Objective To analyze the levels and clinical significances of IL-18,Caspase-3 and nerve tissue-specific protein S-100B at different disease extent and different stages of infants with hypoxicischemic encephalopathy (HIE).Methods This study was clinical experimental studies.Sixty-seven infants with HIE (23 cases of mild HIE,23 cases of moderate HIE,21 cases of severe HIE) from February 2008 to June 2009 in Hebei Children's Hospital were enrolled.The levels of IL-18,Caspase-3 and S-100B protein in all samples were measured at acute phase (1 d,3 d) and recovery phase (7 d) by ELISA method.Twenty healthy full-term neonates were selected as the normal control group.Multi-factor analysis of variance and Pearson correlation test was used for statistical analysis.Results The levels of the three indicators in the moderate and severe group were higher than the normal control group.In the moderate group,IL-18 levels were(132.15 ± 9.87),(150.31 ± 15.04) and (87.91 ± 9.93) ng/L,Caspase-3 levels were (5.79 ±0.64),(7.36 ± 1.57)and (3.79 ±0.61) μg/L,S-100B levels were(6.82 ±0.61),(9.62 ± 1.29) and (10.76 ± 1.64) μg/L.In the severe group,IL-18 levels were (160.23 ± 16.03),(189.86 ± 18.32) and (107.35 ± 13.02) ng/L;Caspase-3 levels were (6.86 ± 1.02),(9.54 ± 1.43) and (5.25 ± 0.71) μg/L;S-100B levels were(8.90 ± 0.32),(12.54 ± 0.89)and(13.53 ± 0.75) μg/L.In the normal control group,IL-18 levels were (71.08 ± 11.52),(72.53 ± 11.05) and (71.93 ± 11.30) ng/L; Caspase-3 levels were (2.84 ± 0.52),(2.98 ± 0.53) and (2.87 ± 0.52) μg/L; S-100B levels were (1.50 ± 0.25),(1.62 ±0.30)and(1.53 ±0.29) μg/L IL-18 levels,Caspase-3 levels and S-100B levels in severe group were higher than the moderate group and the mild group were higher than the mild group.IL-18 levels were (73.46 ± 4.77),(77.59 ± 4.02) and (72.87 ± 6.92) ng/L ; Caspase-3 levels were (3.13 ± 0.31),(3.63±0.40) and (3.26 ±0.45) μg/L;S-100B levels were(3.68 ±0.40),(5.851 ±0.63) and(6.95 ± 0.58) μg/L in the mild group.S-100B levels in the mild group were higher than that in the normal control group.The IL-18 and Caspase-3 levels were risen in the third day to the first day in the acute phase of the moderate group and severe group,decreased in the recovery phase.Serum S-100B protein levels in the acute and recovery phase increased gradually,and there was no correlation between the three indicators (r-=0.321,0.14,0.48,P=0.438,0.974,0.911 respectively).Conclusions IL-18,Caspase-3 and S-100Bwere involved in the pathophysiological process of HIE.The levels were closely related to the severity and disease progression of HIE,the severer of the illness,and the higher of the levels.Dynamic monitoring the changes of the three indicators may contribute to an early diagnosis,condition monitoring and prognosis of HIE.

Objective To detect the expression of advanced glycation end-products (AGEs) in the sun exposure and non-exposure skin,to observe the elastic protein in the same location,and to explore the relationship between non-enzymatic glycation reactions and,photo-aging skin morphological changes.Methods In the exposure and non-exposure specimens from 30 patients,elastic fibers were stained with Gomori staining,and immunohistochemistry for AGEs was performed.Results AGEs expressed clearly positive in all elastic fibers of degeneration markedly (+ + above) of exposure skin,and appeared line-like in markedly(+ + above)elastic fibers.While in all non-exposure skin,AGEs expressed negative.In the exposure skin with middle or old age,compared with non-exposure skin,AGEs expressed more and elastic fibers were hyperplasia with thickening,curling and irregular distribution.The skin parts of AGEs expressed accorded with where elastic fibers degenerated.Conclusions Ultraviolet radiation can induce denaturation of elastic fibers and non-enzymatic glycation may contribute to the mechanism of skin photo-aging.

Objective:To analyze the changes in T lymphocyte subsets, B lymphocytes and NK cells in children with active tuberculosis (TB) and their clinical significance.Methods:T lymphocyte subsets, B lymphocytes and NK cells in peripheral blood samples of 106 patients with acute TB (TB group) and 106 healthy children (healthy control group) were detected by flow cytometry and compared between different groups.Results:The percentages of CD3 + T, CD4 + T and NK cells as well as the CD4 +/CD8 + T cell ratio were significantly lower in the TB group than in the healthy control group ( Z=-3.783, P=0.000; Z=-5.401, P=0.000; Z=-3.434, P=0.001; Z=-2.014, P=0.044). The percentages of double negative T (DNT) and B cells in the TB group were significantly higher than those in the healthy control group ( Z=2.765, P=0.006; Z=6.880, P=0.000). No significant difference in the percentage of CD8 + T or double positive T (DPT) cells was observed between the two groups ( P>0.05). The expression of peripheral lymphocyte subsets varied in TB children of different age groups (0-<3, 3-<6, 6-<10 and 10-<16 years old). There were significant differences in CD3 + T, DNT and B cells among the four age groups ( H=10.081, P=0.018; H=14.583, P=0.002; H=8.498, P=0.037). The percentage of CD4 + T cells was significantly lower in children with extrapulmonary TB than in those with pulmonary TB ( Z=-3.068, P=0.002). No statistically significant difference in other lymphocyte subsets was found between children with extrapulmonary and pulmonary TB ( P>0.05). Conclusions:Tuberculosis could lead to immune dysfunction in children. Dynamic monitoring of the changes in peripheral lymphocyte subsets in children with TB could be conducive to better assessment of immune status and providing personalized treatment.

Objective To prospectively compare cadmium-zinc-telluride (CZT) SPECT gated myocardial perfusion imaging (GMPI),conventional SPECT MPI and cardiac MRI for the assessment of left ventricular volume and ejection fraction in patients with heart failure.Methods From July 2016 to October 2016,a total of 35 patients (27 males,8 females,average age:(52.7±14.9) years) with heart failure were consecutively included.All patients underwent CZT SPECT GMPI,conventional SPECT GMPI and cardiac MRI within 7 d.LVEDV,LVESV and LVEF of three imaging modalities were calculated.One-way analysis of variance,Pearson correlation analysis and Bland-Altman analysis were used.Results CZT SPECT showed excellent correlation with conventional SPECT for LVEDV,LVESV and LVEF (r values:0.983,0.986 and 0.910,respectively;all P＜0.001).Bland-Altman analysis revealed good agreement between CZT SPECT and conventional SPECT for LVEDV,LVESV and LVEF.The correlation between CZT SPECT and cardiac MRI for LVEDV,LVESV and LVEF were all significant (r values:0.864,0.896 and 0.836,respectively;all P＜0.001).Compared with cardiac MRI,CZT SPECT showed systemic underestimation of LVEDV and LVESV and good agreement of LVEF by Bland-Altman analysis.Conclusions CZT SPECT has high clinical value for patients with heart failure.Despite underestimating LVEDV and LVESV,it correlated well with cardiac MRI.It also has a good agreement with conventional SPECT on left ventricular volume and LVEF.

Objective To develop a tetra-primer amplification refractory mutation system PCR(T-ARMS-PCR)assay for detecting four single nucleotide polymorphisms(SNPs): rs1801133,rs1801131, rs1805087 and rs1801394 associated with folate metabolism in a single reaction tube.Methods Methodology was developed.Then, it applied the established method to analyze 150 physical examination children′s anticoagulant blood samples admitted to the Department of Clinical Laboratory, Children′s Hospital of Hebei Province from January 2017 to April 2017.Four sets of chimeric primers consisting of a universal Tag sequence and targeting rs 1801133,rs1801131,rs1805087 and rs1801394 fused to the specific sequence were designed according to the T-ARMS-PCR principle and the multiplex chimeric primers strategy.A single tube PCR was then conducted after optimizing the primer concentrations and the reaction conditions.The amplified products were analyzed by QIAxcel capillary electrophoresis, and the corresponding SNP genotypes of 150 samples were identified.Furthermore,all samples were verified by direct sequencing.And the Hardy-Weinberg Equilibrium(HWE)testing of four SNPs from 150 samples were conducted by SPSS17.0 Chi-square test.Results The improved T-ARMS-PCR combined with capillary electrophoresis can accurately verify eight different alleles of the four SNPs associated with folate metabolism at one time in 3 hours.Four SNPs of 150 whole blood samples were accurately classified and the results were completely consistent with direct sequencing.All the genotype frequencies of these four SNPs were in HWE (χ2rs1801133=0.69, Prs1801133=0.40; χ2rs1801131=0.21, Prs1801131=0.64; χ2rs1805087=3.32, Prs1805087=0.07;χ2rs1801394=1.91, Prs1801394=0.17).Conclusions The proposed improved T-ARMS-PCR combined with capillary electrophoresis in this study can accurately verify four SNPs associated with folate metabolism in a single reaction tube.This method might be a valuable tool to specifically guide the folate supplement in general population.

Objective:To analyze the epidemiological characteristics of respiratory syncytial virus (RSV)-caused respiratory diseases in children in Hebei Province and the changes in peripheral blood lymphocyte subsets and inflammatory indexes.Methods:A total of 9 491 sputum specimens and 9 491 paired peripheral blood specimens were collected from children admitted to Hebei Children′s Hospital for respiratory infection in 2018. RSV-positive sputum specimens were screened by multiple detection reagents for 13 kinds of respiratory pathogens. Flow cytometry was uses to detect T and B lymphocytes and NK cells in peripheral blood samples of randomly screened RSV-positive children. Procalcitonin (PCT) was measured by Roche E411 luminescence analyzer. Hypersensitive C-reactive protein (hs-CRP) was detected by Roche Cobas 8000 C701 biochemical analyzer. White blood cells (WBC) were measured by Sysmex XN-BN3 hematometer.Results:The positive rate of RSV in children with respiratory diseases was 13.08% in Hebei Province in 2018. There were significant differences in RSV-positive rates among different age groups (χ 2=479.297 6, P<0.000 1). The positive rate of RSV decreased gradually with age (χ 2=-20.282 7, P<0.000 1) and was higher in male than in female (χ 2=34.552 7, P<0.000 1). The incidence of co-infection of RSV with other respiratory pathogens was 29.49% (366/1 241), mainly caused by human rhinovirus (HRV, 150/1 241) and adenovirus (ADV, 40/1 241). The main epidemic seasons of RSV infection were winter and spring. The epidemic trends of simple RSV infection and co-infection were consistent. There were significant differences in inflammatory indexes, WBC ( P<0.01), CD4 + ( P=0.015) and CD4 + /CD8 + cells ( P=0.016) between simple RSV infection and co-infection groups. Conclusions:RSV was a common pathogen causing respiratory diseases in children in Hebei Province. The younger the children were, the more likely they were to be infected with RSV. RSV infection was easily complicated by HRV or ADV infection. The epidemic seasons of RSV infection in Hebei were winter and spring. Both simple infection and co-infection of RSV might result in immune dysfunction.

Objective To investigate the effects of mannan-binding lectin(MBL) on TNF-α production induced by peptidoglycan (PGN) and its mechanism in human THP-1/CD14 monocytes.Methods The THP-1/CD14 cells were stimulated for 24 h with PGN at the indicated ratios after pretreated with human natural MBL at concentrations ranging from 1 to 20 mg/L for 2 h.The content of TNF-α and IL-6 in culture supernatants were detected by ELISA,and the levels of TNF-α and IL-6 mRNA expressions in these cells were determined by RT-PCR.FACS was used to investigate the interaction of MBL with THP-1/CD14 cells and the impact of MBL on PGN binding to THP-1/CD14 cells.Western blot was used to detect PGN-induced NF-κB translocation in THP-1/CD14 cells.Results ELISA showed that secretion of TNF-α and IL-6 from THP-1/CD14 cells could be induced by PGN ;The productions of TNF-α and IL-6 by THP-1/CD14 cells induced with PGN were profoundly inhibited by MBL at higher concentrations (10-20 mg/L) but not MBL at lower concentrations (1 mg/L).RT-PCR analysis also indicated that the mRNA expressions of TNF-α and IL-6 in THP-1/CD14 cells were decreased by MBL at higher concentration,compared to the corresponding THP-1/CD14 cells stimulated with PGN only.FACS showed that the binding of MBL to THP-1/CD14 cells was evident in a Ca2+-dependent manner.PGN could competitively inhibit the binding of MBL to THP-1/CD14 cells.MBL could competitively inhibit the binding of PGN to THP-1/CD14 cells by binding to THP-1/CD14 cells directly.Similarly,MBL at higher concentration (20 mg/L) decreased the NF-κB translocation in THP-1/CD14 cells.Conclusion MBL may inhibit TNF-α and IL-6 production induced by PGN in THP-1/CD14 cells through NF-κB signaling pathways,suggesting that MBL can play some roles in the regulation of PGN-induced inflammatory response.

Objective To analyze the iatrogenic fractures following treatment of type 12-A humeral shaft fractures with antegrade intramedullary nails. Methods We reviewed the patients who had been treated with antegrade intramedullary nails for type 12-A humeral shaft fracture from October 2006 to March 2017. They were 13 males and 13 females with an age range from 22 to 76 years and an average age of 44. 5 years. Six of them were complicated with other fractures or radial nerve injury. We divided the patients into an iatrogenic fracture ( IF ) group and a non-iatrogenic fracture ( NIF ) group and compared gender, age, fracture type, reaming and nonunion between the 2 groups. Results The follow-up time for the 26 patients ranged from 13 to 182 weeks ( 63. 8 weeks on average ) . Of them, 7 suffered iatrogenic fracture and 4 nonunion. The union time for the other 22 patients ranged from 9 to 29 weeks ( 14. 6 weeks on average ) . All the 7 patients in the iatrogenic fracture group were treated with reaming while only 9 of the 19 patients in the non-iatrogenic fracture group were treated with reaming. Nonunion occurred in 3 patients in the iatrogenic fracture group but in only one patient in the non-iatrogenic fracture group. All the above comparisons were statistically significant between the 2 groups ( P < 0. 05 ) but there were no significant differences in comparisons between the 2 groups regarding gender, age, nail diameter, fracture type, way of locking proximal and distal screws or rate of secondary surgery ( P > 0. 05 ) . Conclusions Antegrade intramedullary nailing may cause an iatro-genic fracture in the treatment of type12-A humeral shaft fractures. The iatrogenic fracture may affect fracture union. Intraoperative reaming may be the risk factor for the iatrogenic fracture.

Objective To analyze the molecular epidemiology of adenovirus ( ADV) causing acute respiratory diseases and to investigate the mixed infection of ADV and other respiratory pathogens in Hebei Province. Methods Sputum samples were collected from inpatient children with acute respiratory diseases at Children′s Hospital of Hebei Province between June 2017 and May 2018. Multiplex reverse transcription PCR assay was used to detecte 13 kinds of respiratory pathogens. Nested PCR was performed to amplify ADV hexon gene and the amplified products were then sequenced. Results A total of 353 ADV-positive speci-mens were detected in 8839 specimens with a positive rate of 3. 99％. Significant difference in the positive rate of ADV was not observed between male and female patients (χ2=0. 0003, P=0. 99), but found among different age groups (χ2=115. 69, P<0. 001). All isolated ADV strains belonged to 11 serotypes, which were type 1 (16. 15％, 57/353), type 2 (35. 98％, 127/353), type 3 (21. 25％, 75/353), type 4 (1. 13％, 4/353), type 5 (11. 33％, 40/353), type 6 (3. 97％, 14/353), type 7 (8. 22％, 29/353), type 31 (0. 28％, 1/353), type 41 (0. 28％, 1/353), type 55 (0. 28％, 1/353) and type 57 (1. 13％, 4/353). Among the 353 ADV-positive specimens, 259 were mixed infections mainly caused by ADV and human rhinovirus (35. 52％). ADV and respiratory syncytial virus co-infections accounted for 12. 74％ and 33. 20％ of the mixed infections involved three or more pathogens. ADV could be detected throughout the year, especially in September and April to May. The predominant serotypes were types 1, 2 and 3. The av-erage ages of the two groups of ADV infection alone and ADV mixed infection were (27. 56±24. 67) months and (21. 33 ±20. 28) months, respectively, and the difference between them was statistically significant (P=0. 037). The incidence of ADV 2 infection alone was 25. 77％ (25/97), which was lower than that of ADV 2-involved mixed infection [39. 84％ (102/256),χ2=6. 05, P=0. 014]. However, the rate of ADV 7 infection alone was significantly higher than that of ADV 7-involved mixed infection [16. 49％ 16/97) vs 5. 08％ (13/256),χ2=6. 05, P<0. 001]. Conclusion ADV 1, ADV 2 and ADV 3 were the predominant serotypes circulating in Hebei Province from June 2017 to May 2018, especially in September and April to May. The younger the patients were, the higher the incidence would be. ADV 2 was prone to cause mixed infections with other respiratory pathogens, while ADV 7 was less common in mixed infections. Younger pa-tients were more susceptible to mixed infections. The most common co-infection was caused by ADV and hu-man rhinovirus.

Objective To analyze the factors that influence screw penetration after plating for proximal humerus fractures of Neer two-or three-part.Methods We reviewed the patients with proximal humerus fracture of Neer types Ⅱ & Ⅲ who had been treated from January 2006 to June 2017.The data were documented regarding gender,age,Neer classification,osteoporosis,surgical procedure,cephalic screw number,type of medial support and screw penetration.They were divided into 2 groups according to the presence or absence of screw penetration.x2 test and independent samples t test were used with software IBM SPSS 22.0.The multivariate logistic regression was used to identify the independent risk factors when there were two or more statistically significant factors.The two-side test was used and significance was set to P ＜ 0.05.Results Altogether 78 patients were included in our analysis,including 27 males (34.62％) and 51 females (65.38％) with an average age of 57.41 years (from 18 to 85 years).Their average follow-up time was 7.7 months (from 3 to 24 months).There were 54 Neer two-part fractures (69.23％) and 24 Neer three-part ones (30.77％).Screw penetration was observed in 18 patients (23.08％) but not in the other 60 ones (76.92％).The x2 test and independent samples t test revealed that screw penetration was significantly associated with gender (P =0.003),osteoporosis (P =0.003) and cephalic screw number (P =0.003) but not with age (P =0.256),Neer classification(P =0.754),surgical procedure((P =0.308) or type of medial support (P =0.186).The multivariate logistic regression revealed that osteoporosis (P =0.027) and cephalic screw number (P =0.026) were independent risk factors for screw penetration.Conclusions After plating for proximal humerus fractures of Neer two-or three-part,the patients with osteoporosis and those receiving more than 5 cephalic screws may face a higher risk of screw penetration than those without osteoporosis and those receiving 4 cephalic screws.