Objective To study protective immunity effects of co-immunization with P30 DNA vaccine and protein vaccine. Methods Forty-eight 5-6 weeks old BALB/c female mice were divided into four groups (A,B,C,D), 12 mice of each group. In group A (control group) each mouse was immunized with 100 ?g pcDNA3.1 plasmid DNA by intramuscular (i.m.) for three times at week 0,2 and 4; in group B (P30 protein group) each mouse was immunized (i.m.) with 50 ?g rP30+50 ?g CFA for three times at week 0, 2 and 4; in group C (pcDNA3.1-P30 group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for three times at week 0, 2 and 4; in group D (P30 DNA+rP30 co-immunization group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for two times at week 0, 2 and immunized by subcutaneous with 50 ?g rP30+50 ?g CFA at week 4. Each mouse was infected with 100 tachyzoites of Toxoplasma gondii RH strain four weeks later after last immunization. The anti-P30 antibodies were detected with ELISA before the challenge. Results The P30 DNA vaccine was successfully constructed. High titers of anti-P30 antibodies were induced in each mouse immunized with DNA vaccine. The protective trial proved that there was no significant difference between control group and experimental group though the survival time of mouse from experimental group had been prolonged. Conclusion The P30 DNA vaccine could induced high titers of anti- P30 antibodies in immunized mice, and it may be a potential DNA vaccine candidate.

Objective To evaluate efficacy of routine cleaning and disinfection methods for incubators,and put forward a feasible improvement solution.Methods 30 incubators used in a neonatal intensive care unit of a hospital between Decem-ber 2013 and June 2014 were chosen and randomly divided into baseline,control,and trial groups(10 incubators in each group).Baseline group and control group were disinfected by routing disinfection method (wiping internal and external sur-faces of incubators with water and chlorine-containing disinfectant),trial group adopted intensified disinfection method (wi-ping internal surfaces of incubators with alcohol)on the basis of routine disinfection,disinfectant efficacy of three groups were compared.Results In baseline group,unqualified incubators were initially detected on the fourth day of monitoring, all incubators were contaminated in varying degrees on the seventh day of monitoring,the detection rate of unqualified spec-imens was 31.43% (88/280).The median time for the initial detection of unqualified incubators in control group and trial group were on the fifth day and seventh day respectively,there was significant difference between two groups(χ2 =12.38, P <0.05);The unqualified rate of trial group was significantly lower than control group (15.36%[43/280]vs 32.86%[92/280],χ2 =23.43,P <0.05 ).Conclusion Intensified disinfection with alcohol on the basis of routine disinfection method can effectively improve the disinfectant efficacy of the surface of incubators,it is convenient,inexpensive and safe, and worth to be popularized in primary hospitals.

OBJECTIVE:To investigate the preparation technic and method of quality control of Xinyoukexing tincture(XYKX) METHODS:To determine the content of Valaciclovir by spectrophotography,and podophyllotoxin by dual-wavelength spectrophotography RESULTS:In the range of 14 88～34 72?g/ml,the Valaciclovir concentration in XYKX was in direct proportion to the absorption at 310 4nm,C=44 5 905A+1 7 556,r=0 9 998 In the range of 17 50～50 12?g/ml,the podophyllotoxin concentration in XYKX was in direct proportion to the absorption at 292nm,C=97 7 981Ap292+1 3 614,r=0 9 999,AP292=As292-As325 CONCLUSION:The preparation technic of XYKX is simple and the method of quality control is feasible

Objective To investigate the changes of plasma adrenodullin(ADM) and C-type natriuretic peptide(CNP) level in children with chronic heart failure(CHF) and its clinical implications.Methods Forty-two children with CHF were collected.The patients suffered from dilated cardiomyopathy,congenital heart defects,and other heart diseases.According to a modified scoring system described by Ross and Reithman,16 patients were classified as class Ⅱ,14 as class Ⅲ,and 12 as class Ⅳ.Plasma levels of ADM and CNP were measured by radioimmunoassay assay in these patients and 11 healthy children.Echocardiography was performed to measure left ventricular function and the ratio of E/A.Results Plasma ADM and CNP levels of CHF patients were significantly elevated as compared with those of the control subjects [(218.27?106.53) ng/L vs(74.39?53.99) ng/L,P=0;(190.27?108.38) ng/L vs(92.59?(59.46) ng/L),P0.05).ADM levels were elevated with the advancing severity of CHF determined by a modified scoring system described by Ross.However,the plasma CNP levels in the normal state wasn′t significantly different from those observed in class Ⅱ.Likewise,the plasma CNP level in the class Ⅲ was not significantly different from that observed in class Ⅱ.Conclusions ADM and CNP might play a compensatory and defensive roles in the pathophysiology of the pediatric CHF.ADM may be a biochemical marker for evaluation the severity of the chronic heart failure in children,but also a new prognostic indicator of this syndrome.

Objective To collect the families with generalized epilepsy with febrile seizures plus(GEFS+) and analyze the clinical status and heredity features of Chinese GEFS+.Voltaged-gated sodium channel ?1 subunit(SCN1B) gene of 2 families were detected,and expect to find new mutation sites.Methods All participant in the study of 2 families members were informed of voluntary participate in this investigation,health examination and blood sampling.All 6 gene exons of proband,patients and healthy control group were sequenced.The sequencing result was compared and analyzed with the normal sequence of genomic exon fragment and exon fragment sequencing result of control group through internet(BLAST).Results 1.A new G/A heterozygous polymorphism (G181A)was found in the 181th basyl of SCN1B gene exon 3,and codon was changed from TCG to TCA,both encoding serine (Ser,S).It was synonymous mutation.2.A new G/A heterozygous polymophism(G15A)was found in the 15th basyl of SCN1B gene exon 3,and codon was changed from GAG to GAA,both encoding glutamic acid(Glu,E).It belonged to synonymous mutation.3.A new T/C heterozygous polymorphism (T37C)was found in the 37th basyl of SCN1B gene exon 6.The patients genetype were:5 cases with T/C heterozygote,3 cases with T/T homozygote,2 cases with C/C homozygote.Healthy control group were all T/T homozygote.Allele frequency distribution for T was 55.0%,and 45.0% for C.4.A new A/C heterozygous polymorphism (A81C)was found in the 81th basyl of SCN1B gene exon 6.The patients genetype were:5 cases with A/C heterozygote,3 cases with A/A homozygote,2 cases with C/C homozygote.Healthy control group were all A/A homozygote.Allele frequency distribution for A was 55.0%,and 45.0% for C.Conclusions Two new heterozygous polymorphism (G181A),(G15A) were found in SCN1B gene exon 3.Two new heterozygous polymorphism (T37C),(A81C) were found in SCN1B gene exon 6.These 4 polymorphism enriched single nucleotide polymorphism(SPN) database and provided candidate sites for the research of epilepsy susceptbility polymorphisms.

Generalized epilepsy with febrile seizures plus(GEFS+) is a new epilepsy syndrome proposed by International League Against Epilepsy.At present,the progress of genetic studies of GEFS+ focus on gene mapping based on family analysis,many researches indicate that GEFS+ is associated with voltaged-gated sodium channel(SCN) gene mutation.This paper intends to discuss the relationship beween GEFS+ and SCN1B,SCN2B,SCN1A,SCN2A genes,mutations in order to improve the cognition about GEFS+.

Objective To set a rapid,simple gene diagnosis method for Down syndrome.Methods Three short tandem repeats(D21S11,D21S1270,D21S1437)loci in or near Down syndrome critical region(DSCR) were analyzed and detected by polymerase chain reaction and DNA quantitative analysis in 11 core ancestry.Results There were four types by DNA quantitative analysis to different individuals at a short tandem repeats(STR) locus.In type one,a homozygote of one allelic gene was detected.In type two,a normal heterozygote of two allelic genes was found,the content or two DNA electrophoresis bands was approximately 1∶1.In type three,a Down syndrome patient of two allelic genes was discovered.The quantity of two electrophoresis bands was nearly 2∶1.In type four,the patient showed three DNA electrophoresis bands which the content was approximately 1∶1∶1.Conclusion A rapid gene diagnosis and prenatal diagnosis method for Down syndrome can be used for quantitative analysis of STR polymorphism loci.

Objective To explore the mutation of voltage-gated sodium channel ?1 subunit(SCN1B)gene in Chinese Han families with generalized epilepsy with febrile seizures plus(GEFS+).Methods Two families pedigrees were established and disease history,physical examination were collected in order to analyze the mode of inheritance.The mutation sites of reported SCN1B gene exon 3-(C46T,G47A,C156G)were detected by polymerase chain reaction sequence special primers(PCR-SSP)method after genomic DNA were extracted.Results There were 13 patients in the 2 families,all were Han people and 9 cases were living.The results showed the mode of inheritance basically corresponds with autosomal dominant inheritance complicated with incomplete penetrance,and leaded to different kinds of phenotype.The mutation sites of SCN1B gene exon 3 were detected by using PCR-SSP method,and heterozygote was not found,and point mutations were also not found in the 2 families.Conclusions GEFS+ is a complex disorder with genetic heterogeneity.There were no gene mutations of SCNIB in the 2 GEFS+ families,which might suggets the possibility of insufficient samples as the patients came from Henan province and the possibility of differences from races and regions of other countries.

ObjectiveTo study the effect of pre-arrest and post-arrest mild hypothermia after restoration of spontaneous circulation (ROSC) on myocardial function, ultrastructure, apoptosis of myocardial cells in rabbits with ventricular fibrillation.Methods Sixty-two male New Zealand rabbits were randomly allocated into five groups: namely normothermic control group (NTC group,n = 10), hypothermia control group (HTC group,n = 10), normothermic resuscitation group (NTR group,n = 14), hypothermia pre-arrest group (HPRA group,n = 14), and hypothermia post-arrest group (HPOA group,n = 14). The normal temperature was controlled at (39.0±0.5)℃, and the hypothermia (33.5±0.5)℃. Ventricular fibrillation cardiac arrest (CA) was reproduced in rabbits by transcutaneous epicardium electrical stimulation. The parameters of hemodynamics were monitored dynamically for 4 hours in all the groups, including heart rate (HR), left ventricular end diastolic and systolic pressure (LVEDP/LVESP), maximal rate of increase/decrease in left ventricular pressure (±dp/dt max), and mean arterial pressure (MAP). The body temperature of rabbits in hypothermia groups was maintained by surface cooling for 4 hours followed by rewarming. The survived rabbits were sacrificed at 48 hours after resuscitation, and myocardial apical tissue was harvested for observation of ultrastructure with electronic microscope, and to observe apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining.Results① Resuscitation investigation: there was no significant difference in rate of ROSC, time of CPR and energy of defibrillation among HPRA, HPOA, and NTR groups [rate of ROSC: 85.71%, 71.43%, 71.43%; time of CPR (seconds): 45.3±30.2, 61.2±41.3, 82.3±63.8;energy of defibrillation (J): 14.3±8.9, 22.0±15.5, 25.0±15.8, allP> 0.05].② Hemodynamics: compared with normal temperature groups, animals in hypothermia groups exhibited lower levels of HR (allP< 0.05). Compared with NTR group, HPRA group exhibited higher levels of LVESP (mmHg, 1 mmHg = 0.133 kPa) at 0.5, 1, 2 and 3 hours post ROSC (0.5 hour: 103.8±14.3 vs. 91.6±13.3, 1 hour: 107.2±14.1 vs. 82.7±8.5, 2 hours: 109.0±16.9 vs. 88.8±12.9, 3 hours: 109.1±14.6 vs. 89.3±14.3, allP< 0.05). Compared with NTR group and HPOA group, HPRA group exhibited lower levels of LVEDP (mmHg) at 0.5 hour post ROSC (3.70±0.85 vs. 7.61±2.73, 7.02±3.12, both P< 0.05). Compared with NTR group, HPRA group exhibited lower levels of LVEDP at 1 hour post ROSC (4.34±1.44 vs. 6.99±1.96,P< 0.05). In HPRA group, the level of+dp/dt max (mmHg/s) was higher than that of NTR group and HPOA group at 1 hour and 2 hours post ROSC (1 hour: 2 759.5±321.6 vs. 2 123.0±304.5, 2 283.7±234.2, 2 hours:2 730.6±425.1 vs. 2 221.5±392.9, 2 252.6±476.0, allP< 0.05). There were no significant differences in -dp/dt max and MAP levels among three CPR groups.③ The survival rate at 48 hours post ROSC of NTR, HPRA and HPOA groups was 60%, 75%, and 100%, respectively. Compared with NTR group, higher survival rate was found in HPOA group at 48 hour post ROSC (P< 0.05).④ Compared with NTR group, less damage to myocardial ultrastructure was found in HPRA and HPOA groups. Apoptosis index (AI) was lower in HPRA and HPOA groups than that in NTR group [(28.05±9.82) %, (26.39±8.98) % vs. (42.02±13.36) %, bothP< 0.05].Conclusions Our study shows that mild hypothermia has no effect on ROSC rate. Pre-arrest hypothermia can ameliorate myocardial systolic function of rabbit in early stage after ROSC, and it has no negative influence on diastolic function. Post-arrest mild hypothermia produces no negative influence on myocardial function of rabbit, but it improves 48 hours survival rate in ROSC rabbits. Both pre-arrest and post-arrest mild hypothermia therapy can attenuate myocardial injury in CA model of rabbits by ameliorating mitochondrial injuries and suppressing apoptosis of myocardial cells.