Animals ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; metabolism ; virology ; Myocardium ; metabolism ; Osteopontin ; metabolism ; Virus Diseases ; metabolism

ObjectiveThe purpose of the study was to summarize the characteristics of cellular and humoral immunity in children with Kawasaki's disease (KD),and to explore the role of cellular and humoral immunity in the pathogenesis of KD.MethodsOne hundred and ninety-six subjects with KD were diagnosed and observed.The serum IgG,IgM,IgA and C3,C4 levels were detected by velocity scatter turbidimetry.The levels of T lymphocyte subsets,natural killer cells (NK cells),and B cells were analyzed by flow cytometry.Chi-square test,independent t-test and Spearman analysis were adopted to analyze data.ResultsOne hundred and seventy-two cases were diagnosed as typical KD,24 cases were incomplete KD,17 cases were refractory KD,179 cases responded to IVIG therapy,and 34 cases were CAL.The occurrence of coronary artery lesion (CAL) in the refractory KD group was 52.9％,which was evidently higher than those patients responding to IVIG therapy(14.0％,P＜0.05).In the group of KD,the level of CD19 absolute value(1165±556)/μl was higher than that in the control group(P＜0.05),the ratio of CD4/CD8 (2.19±0.77) was higher than that in the control group (P＜0.05),the levels of serum IgM ( 1.41 ±0.45) g/L,IgA (0.81 ±0.35) g/L,C3 ( 1.31 ±0.26) g/L were higher than those in the control group (P＜0.05),the levels of CD8 (671±308)/μl,NK (337±233)/μl absolute value were lower than those in thc control group (P＜0.05).In refractory KD subgroup,the levels of CD19 absolute value,serum IgM,C3 and the ratio of CD4/CD8 were higher than those in the subgroup responding to IVIG therapy(P＜0.05),while the levels of CD8,NK absolute value were lower than those in the latter(P＜0.05).In the CAL subgroup,the levels of CD19 absolute value,serum IgM,IgA and the ratio of CD4/CD8 were higher than those in the non-CAL subgroup (P＜0.05),while the levels of CD8,NK absolute value were lower than those in the latter(P＜0.05).The differences of all data between typical KD subgroup and incomplete KD subgroup were insignificant (P＞0.05).The severity grade of CAL was positively correlated with the ratio of CD4/CD8(P＜0.05).ConclusionThere are cellular immunity and humoral immunity disturbances in the acute stage of KD,while cellular immunity disturbances are more striking.Cellular immunity and humoral immunity are all involved in the pathogenesis of KD.The immunity dysfunction is more significant in refractory KD and CAL.The occurrence rate of CAL is high in refractory KD.The ratio of CD4/CD8 is relevant to the severity of vascular injury.

BACKGROUND:Our previous study has already manifested that Chinese medicine Xiaokening can effectively prevent and treat the early diabetic nephropathy. OBJECTIVE:To compare the effect of rhein and archen on the apoptosis of mesangial cells of rats cultured with high glucose. METHODS:Mesangial cells were stimulated by different concentrations of rhein and archen (20, 40 and 80 μmol/L). Morphology of apoptotic cells was observed by hematoxylin-eosin staining. Karyon apoptosis was examined by fluorescence staining of DAPI. cellapoptosis rate was detected by flow cytometry. RESULTS AND CONCLUSION:The outcome of hematoxylin-eosin staining and DAPI staining indicated that the effect of rhein was much stronger than that of archen on the apoptosis of mesangial cells in the rats cultured with high glucose. Comparison of the apoptosis rate also indicated that the rhein had a stronger effect on the earlier and the later stage apoptotic rate of mesangial cells than archen. Both rhein and archen with different concentrations can induce apoptosis of mesangial cells, but the effect of rhein is much stronger.

Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.

Objective To investigate the detection of human bocavirus (HBoV) in children with acute respiratory infection and to explore the relationship between viral load and clinical characteristics of acute respiratory infection in children.Methods A total of 4 501 nasopharyngeal secretion samples were collected from hospitalized children with acute respiratory infection from January 2013 to June 2013.HBoV-positive children were divided into simple infection group and mixed infection group.Children with HBoV DNA≥1 × 104 copy/mL were categorized into high viral load group,while those with HBoV DNA ＜1 × 104 copy/mL were categorized into low viral load group.HBoV was determined by fluorescence quantitative polymerase chain reaction (PCR).Respiratory syncytial virus (RSV),influenza virus (Inf)-A,Inf-B,parainfluenza virus (Pinf)-Ⅰ 、Pinf-Ⅱ 、Pinf-Ⅲ and adeno virus antigen were detected by direct antigen-specific immunofluorescence assays.Mycoplasm Pnuemonia was detected by real-time fluorescence quantitative PCR.Serum mycoplasma antibodies were detected by enzyme-linked immunosorbent assay (ELISA).Bacteria was detected by sputum culture.Over the same period,23 children undergoing elective inguinal hernia operation with no respiratory infection or fever were considered as control group.The percentage of peripheral blood T lymphocyte subsets were tested by flow cytometry.Inter-group differences were compared using Chi-square test or Fisher exact test.Viral loads were compared using Mann-Whitney test.Results Two hundred and twenty-two HBoV-positive cases were detected with a positive rate of 5.41％ (222/4 105),33.33％ (74/222) of which were with high viral load and 66.67％ (148/222) were with low viral load.There was a high incidence in the age group of 1-2 years.The simple HBoV infection accounted for 24.32％,including 26 cases with high viral load and 28 cases with low viral load.Wheezing was more common in patients with high viral load than those with low viral load,and the difference was statistically significant (88.46 ％ vs 42.86 ％,x2 =12.295,P=0.001).Among the 222 HBoV-positive cases,the median viral load of HBoV in simple infection group was 3.86 × 103 copy/mL,and 1.0× 103 copy/mL in mixed infection group.The difference of the viral load between these two groups was statistically significant (Z =2.906,P =0.004).Mycoplasma and Streptococcus pneumonia were most commonly detected in the 168 patients with mixed infection.Percentages of CD3+ and CD3+/CD8+ subsets were significantly lower in HBoV simple infection group and mixed infection group,compared to control group (both P＜0.05).However,percentages of CD3 /CD19+,CD19+/ CD23+ subsets were significantly higher in HBoV simple infection group and mixed infection group,compared to control group (both P＜0.05).Conclusions HBoV is one of the pathogens causing acute respiratory tract infection in children,which lead to cellular immunity dysfunction in children.Moreover,children with higher HBoV load are more likely to develop wheezing.Co-infection with other pathogens should be considered in children with low HBoV load.

Objective To evaluate the significance of routine blood test and C‐reaction protein(CRP) ,procalcitonin(PCT ) in early diagnosis of neonatal septicemia .Methods Totally 95 cases of confirmed septicemia ,158 cases of the clinical diagnosis of sep‐ticemia and 160 cases of non‐septicemia who were admitted to Newborn Diagnosis and Treatment Center of Children′s Hospital of Chongqing Medical University from January 2013 to May 2014 were enrolled for this study .The routine blood test ,CRP and PCT concentration of the three groups were analyzed .Taking blood culture as the gold standard ,receiver operating characterristic (ROC) curve was drawn ,sensitivity and specificity of single or combined indicator in early diagnosis of septicemia was analysed .Results There was a significant difference between confirmed septicemia group and non‐septicemia group in PCT ,CRP ,WBC ,I/T ,PLT (P< 0 .05) ;the area under the ROC curve(AUC) was 0 .892 ,0 .847 ,0 .828 ,0 .700 ,0 .681 ,respectively ;the five indicators for con‐firmed the diagnosis positive rate of septicemia group and non‐septicemia group was statistically significant (P < 0 .05) ;the single indicator PCT had better diagnosis value ,its sensitivity and specificity was 84 .2% and 66 .9% .With the application of regression of binary Logistic the AUC of combining PCT ,CRP ,WBC was 0 .974 ,its sensitivity and specificity was 94 .7% and 87 .5% ,which was higher diagnosis value than the only one indicator alone .Conclusion The combination of PCT ,CRP and WBC have contribute to early diagnosis of neonatal septicemia .

Objective;To explore the relationship between exposure to violence and behavior problem among rural middle school students.Methods;A cross-section study was conducted in 3620 middle school students randomly selected from rural area.A questionnaire adapted from WHO Health and Life Experience investigation and Achenbach Children Behavior Checklist(CBCL) were used to measure the exposure to violence and behavior problems respectively.Results;Of the 3620 respondents,the prevalence rate of exposure to violence in past year was 33.8%(1225/3620).The prevalence rate of behavior problems averaged 14.3%(519/3620),and students victimized reported significantly more behavior problems than those no-victimized(22.4% vs 10.9%,P

The effect of inhibition of proliferation of a novel human megakaryoblastic leukemia cell line (HIMeg) by two physiological factors, 1.25-dihydroxyvitamin D3 [1,25(OH)2 D3] and 13-cis-retinoic acid (RA), was investigated. At the range of 10-9 - 10-6 mol/L, 1,25(OH)2 D3 and RA showed significant inhibition of proliferation of the megakaryoblastic leukemia cells,which was demonstrated by count of survival cells,incorporation of [3H]-TdR and [3H]-UR, and cloning efficiency. From above, it can further explain the m.echanism of differentiation-inducers and the effect of 1,25(OH)2D3 on myelofibrosis. It is possible for 1,25(OH)2D3 and RA to be used for a new -treatment.

Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.

Objective Tracking of the vaginal microflora recovery and the expression of immune factors from untreated and treated patients with bacterial vaginosis (BV) by using different administration regimens and studying the relationship of treatment results and regimen selections. Methods 25 healthy females were selected as a control group and 100 BV patients were randomly divided into 4 groups (n=25/group). Group A: Intravaginal administration of metronidazole (× 7 d), Group B:Continuous intravaginal administration of metronidazole (× 7 d) and then live Lactobacillus Capsule (× 7 d) , Group C: Intravaginal administration of nifuratel (× 7 d), Group D: Continuous intravaginal administration of nifuratel (× 7 d) and then live Lactobacillus Capsule (×7 d). The microecological assessment system and EILSA were used to compare the clinical efficacy, vaginal microflora recovery and the changes in IL-8, TLR2 and TNF-αof the vaginal lavage fluid in healthy women or patients with bacterial vaginosis before and after treatments by four treatment strategies. Results ① The vaginal microflora imbalance, flora disturbance, pH value increased were presented in BV group compared with the control group.②Compared to the median of IL-8, TLR2 and TNF-α in vaginal lavage fluids of control group, there was no significant difference in IL-8 level but both TLR2 and TNF-αwere significantly increased (P<0.05) in BV group. The immune factors had no significantly difference in all BV groups.③The therapeutic effect in each BV groups was compared after stopping treatment for 7 days. The cure rate and the vaginal microflora recovery rate were significant higher in group B and D than group A and C (P<0.05). ④ After treatment there was no significant change in IL-8 level but there was an obviouslydecrease in TLR2 and TNF-α(P<0.05). The decreased levels are more significant in groups B and D than groups A and C (P<0.05). Conclusion By combining with the microecological assessment system to evaluate the therapeutic effect of BV, our research suggests that the sequence schemes of nifuratel plus live Lactobacillus Capsule is more effective in therapy effect, restoring normal vaginal micro-ecological environment and vaginal local immunity than metronidazole used alone.