Objective To explore the early surgical treatment for sleep apnea in children with the influence of mental development,provide important basis for children with sleep apnea early surgery.Methods 40 children with sleep apnea underwent bilateral amygdala and/or adenoidectomy (observation group).The Chinese wechsler intelligence scale for children (C-WYCSI) was used for preoperative and postoperative 6 months of intelligence tests,including language IQ and operation IQ test two parts,a total of 11 points.Operation IQ but also contain visual analysis test(VA) and two categories,including geometry test (GD).According to the 1 ∶ 1 matching selection of the same gender,and age(± 3 months),family condition was quite healthy check-up preschool children comparing 40 cases as control group.Results Observation group preoperative total intelligence quotient (IQ),verbal IQ,IQ operation was 82,81,83,respectively,which were lower than those of the control group 101,101,101 (t =4.131,3.952,3.842,all P ＜ 0.05).Postoperative total intelligence quotient(IQ),verbal IQ and operation IQ 98,98,99,respectively,were in normal range,with no statistically significant difference were observed in the control group (all P ＞ 0.05).Conclusion Children with sleep apnea early surgical treatment can improve children mental development.Children with sleep apnea early,breathing and surgical indications should be early surgical treatment.

Objective To explore the efficacy and safety of low-dose ketamine for prevention of shivering during cesarean section under subarachnoid space and epidural anesthesia.Methods Ninety pregnant women who scheduled cesarean delivery under subarachnoid space and epidural anesthesia were randomized divided into three groups:control group (30 patients) low-dose ketamine group (30 patients) and high-dose ketamine group (30 patients).0.9％ sodium chloride,0.25 mg/kg(3 ml),and 0.50 mg/kg (3 ml) ketamine were given in three groups before operation.The incidence of shivering and adverse effect were recorded respectively before anesthesia,after anesthesia 15 min and 30 min and after operation 1 h.Results The level of mean arterial pressure (MAP) in control group after anesthesia 15 main and 30 min and after operation 1 h[(62 ± 10),(58 ± 8),(61 ± 11) mm Hg(1 mm Hg =0.133 kPa)] were significantly lower than those in high-dose ketamine group [(78 ± 12),(82 ± 8),(76 ± 11)mm Hg] and low-dose ketamine group [(72 ± 10),(76 ± 6),(80 ± 7) mm Hg],there was significant difference (P ＜ 0.05).There was no shivering need treat in low-dose ketamine group and high-dose ketamine group.The rate of hallucinations and nystagmus in low-dose ketamine group [0,6.7％ (2/30)] was significantly lower than that in high-dose ketamine group [20.0％ (6/30),50.0％ (15/30)],there was significant difference (P ＜ 0.05).Conclusion The pretreatment with low-dose ketamine on shivering during cesarean section under subarachnoid space and epidural anesthesia has better preventive effectiveness,0.25 mg/kg of ketamine is more effective than 0.50mg/kg.

Objective To analyze the immunogenicity of selected B-cell epitopes of Epstein-Barr virus (EBV) latent membrane protein-2 (LMP2). Methods Three potential dominant B-cell epitopes of LMP2199-209, LMP2318-322 and LMP2381-391 from EBV LMP2 had been predicted using bioinformaties methods. The gene fragments of three epitopes were cloned respectively into pET32a(+) vector and transformed into E. coli strain BL21 (DE3). After identification by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, the expression products were purified by Ni-NTA agarose affinity chromatography. BALB/c mice in immunized groups were immunized by multi-point intracutaneous injection with the three purified epitope proteins,respectively; and mice in control groups were injected with pET32a (+) protein or phosphate buffered saline(PBS), respectively. The sera from mice at week O, week 3 and week 6 of injection were collected for determination of epitope-specific antibody IgG by enzyme linked immunosorbent assay (ELISA) using epitope proteins as coating antigens. The ability of serum antibody recognizing nature EBV antigen was determined at week 6 of immunization. Results Three epitope proteins of LMP2199-209 ,LMP2318-322 and LMP2381-391 were successfully expressed in prokaryotic system. Epitopespecific antibodies IgG could be detected respectively in the sera of all immunized mice, and the levels of antibodies increased with immunized time increasing. The antibody levels in LMP2318-322 immunized group at week 3 and week 6 were significantly higher than that of pET32a (+) protein control group (F= 493.85 and 773.99, respectively; both P＜0. 05), and the antibody levels in LMP2381-391 immunized group at week 3 and week 6 were also significantly higher than that of pET32a (+) protein control group (F= 926.33 and 309.14, respectively; both P＜0.05). Antibody level in LMP2199-209 immunized group at week 6 was significantly higher than that of pET32a ( + ) protein control group (F=87.27, P＜0.05). The antibody IgG in serum from immunized mice with three epitope proteins could all recognize nature EBV antigens, especially LMP2199-209 and LMP2381-391 immunized groups.Conclusions Three possible dominant epitopes of LMP2199-209, LMP2318-322 and LMP2381-391 from EBV LMP2 are prepared by prokaryotic expression system and exhibit obvious immunogenicity, which could be used for further research of EBV infection and related tumor vaccine.

Objective To evaluate the effect of morphine on the growth of subcutaneous tumor of human gastric cancer cells in nude mice.Methods Thirty SPF male BALB/C nude mice,aged 4-5 weeks,weighing 15-20 g,in which the model of subcutaneous tumor of human gastric cancer cell line MGC-803 was established,were randomly divided into 3 groups (n=10 each) using a random number table:control group (group C),normal saline group (group N),and morphine group (group M).The mice in group C received no treatment.Morphine 20 mg/kg was intraperitoneally injected once a day for 14 consecutive days in group M,while normal saline 1.5 ml/kg was given instead of morphine in group N.The caliper was used to measure the tumor size every 2 days starting from 3 days after beginning of administration,and the relative tumor volume was calculated.The nude mice were sacrificed on 15th day,and the tumor tissues were obtained for determination of nuclear factor-kappa B activity and Bcl-2 and Bax protein and mRNA expression by real-time reverse transcriptase polymerase chain reaction and Western blot.Results Compared with group C,the relative tumor volume was significantly decreased,the activity of nuclear factor-kappa B in tumor tissues was significantly decreased,the expression of Bcl-2 protein and mRNA was significantly down-regulated,and the expression of Bax protein and mRNA was significantly up-regulated at each time point in group M (P＜0.05),and no significant change was found in each parameter mentioned above in group N (P＞0.05).Conclusion Morphine can inhibit the growth of subcutaneous tumor of human gastric cancer cells in nude mice,and the mechanism is associated with promotion of apoptosis in tumor cells.

To investigate the effects of particle size, mPEG molecular weight, coating density and zeta potential of monomethoxyl poly(ethylene glycol)-poly(lactic-co-glycolic acid) (mPEG-PLGA) nanoparticles on their transportation across the rat nasal mucosa, mPEG-PLGA-NPs with different mPEG molecular weights (M(r) 1 000, 2 000) and coating density (0, 5%, 10%, 15%) and chitosan coated PLGA-NP, which loaded coumarin-6 as fluorescent marker, were prepared with the nanoprecipitation method and emulsion-solvent evaporation method, and determine their particle size, zeta potential, the efficiency of fluorescent labeling, in vitro leakage rate and the stability with the lysozyme were determined. The effects of physical and chemical properties on the transmucosal transport of the fluorescent nanoparticles were investigated by confocal laser scanning microscopy (CLSM). The result showed that the size of nanoparticles prepared with nanoprecipitation method varied between 120 and 200 nm; the size of nanoparticles prepared with emulsion-solvent evaporation method varied between 420 and 450 nm. Nanoparticles dispersed uniformly; the zeta potential of PLGA-NPs was negative; mPEG-PLGA-NPs was close to neutral; chitosan coated PLGA-NPs was positive; and the efficiency of fluorescent labeling were higher than 80%. In vitro leak was less than 5% within 4 h and nanoparticles were basically stable with lysozyme. The CLSM results show that the transportation efficiency of mPEG-PLGA-NPs with a high PEG coating density and high mPEG molecular weight was significantly higher than that of uncoated PLGA nanoparticles and also that of chitosan coated PLGA-NPs (P < 0.05). The hydrophilcity, zeta potential and particle size of nanoparticles play important roles on the efficiency of mPEG-PLGA nanoparticles to transport across the rat nasal mucosa.

Objectives To investigate the mutational characteristics of MSX1 and PAX9 genes in a family affected by non-syndromic oligodonti so as to study the pathogenesis of oligodontia from a molecular prospective. Methods A family with oligodontia, but of different descent and unrelated healthy controls were enrolled in our study. Genomic DNA was isolated from the blood samples. Mutation analyses were performed by amplifying MSX1 and PAX9 exons and sequencing the products. Results DNA sequencing revealed a novel missense mutation c.348C>T in a highly conserved homeobox sequence of MSX1 and a known polymorphisms c. 469+35- c.469+45del in exon 1 and in intron in the two patients and in two unrelated healthy controls. But we did not detect any mutation in PAX9. Conclusion Our finding suggests the samesense mutation (c.348C>T) and the polymorphisms (c.469+35- c.469+45del) may be responsible for oligodontia phenotype in this Chinese family.

The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.

Objective To explore the HLA-A2 restriction and immunogenicity of 5 previously identified HCV-speeific CTL epitopes. Methods Based on T2 cell, to explore the HLA-A2 restriction of previously identified HCV-specific CTL epitopes by MHC-peptide complex stabilization assay;To detect pep-tide-specific CTL in HLA-A2~+ PBMC stimulated by HLA-A2-restricted peptides by intracellular cytokine staining(ICS) and ELISPOT; To explore the cytotoxicity of peptide-specific CTL to same peptide-loaded T2 cells (target cells) by CTL cytotoxicity test. Results Among 5 previously identified CTL epitopes NS4b_78 (SMMAFSAAL) and NS5a_367 (TVSSALAEL) have high-affinity for HLA-A2 molecules(FI 1) ;ELISPOT results shown that NS4b_78(SMMAFSAAL) and NSSa_367(TVSSALAEL) induced high levels of IFN-γ-se-creting cells [(60±6) SFC/10~4 PBMC vs (4±1 ) SFC/10~4 PBMC, P < 0.01 ; (10 ± 3 ) SFC/10~4 PBMC vs (2±1 ) SFC/10~4 PBMC, P <0.01, respectively] ;ICS results indicated that there were high percentages of CD8~+ IFN-γ~+ T cells in total CD8~+T cells stimulated by these peptides [(2.33 ±0.22 ) % vs (0.05±0.01)%, P <0.001 ; (0.36±0.06)% vs (0.03±0.01)%, P <0.001, respectively]. Furthermore,peptide-specific CTL could effectively kill same peptide-loadcd T2 cells. Conclusion NS4b_78 (SMMAF-SAAL) and NSSa_367 (TVSSALAEL) were identified as HLA-A2-restricted CTL epitopes which could in-duce immune response in vitro.

Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.

Objective To investigate Helicobacter pylori(Hp)infection and HLA-DQA1 allelic frequency in family members of children with recurrent abdominal pain.Methods One hundred and eighteen family members of 20 children with recurrent abdominal pain were divided into two groups:with and without recurrent abdominal pain.Serum Hp antibody was tested by dot immunogold filtration assay and immunophenotyping was determined by Western blot(immunobiot)technique.Polymerase chain reactionsequence specific primers(PCR-SSP)technique Was applied to identify HLA-DQAi allelic frequencies.Hardy-Weinberg equilibrium test was performed(P>0.05),and Chi-square test was used to compare the frequency of HLA-DQA1 alleles between the groups.Results The Hp seropositive rate in 118 members Was 100%and the Hp immunophenotyping was 96.6%.The prevalence of Hp Ⅰ and Ⅱ type was 55.1%(65/118)and41.5%(49/118).HLA-DQA1*0302 allelic frequency Was significantly higher in subjects with recurrent abdominal pain than that in subjects without one(23%vs.2%,X2=13.277,P=0.000).Conclusion There is immunogenetic difference between familial members with and without recurrent abdominal pain infected by Hp,and HLA-DQA1*0302 may be the associated gene contributing to different clinical outcomes after Hp infections.