0.05).The correlation of reduction of intra-disc pressure and accumulation of ablation time was positive.Compared with intra-disc pressure in control group,the pressure change of all ablation groups was statistically significant(P

Objective To investigate the effect of different doses of lidocaine on expression of the high mobility group box 1 protein (HMGB1) in small intestine in septic rats.Methods Fifty adult male Wistar rats weighing 200-250 g were randomly divided into 5 groups ( n =10 each):sham operation group (group S),sepsis group( group Sep ) and different doses of lidocaine group (group L1～3 ).Group S were not applied cecal ligation and puncture (CLP).Sepsis intestinal damage model was performed in group Sep by CLP.Group L1～3 were given intraperitoneally lidocaine in a dose ofS,10 and 20 mg/kg at immediately,1 and 2 h after CLP,respectively.Isometric normal saline was given intraperitoneally in group S and group Sep.The small intestine tissues were taken at 24 and 48 h after CLP.The small intestine morphology was observed with optical microscope.The expression of the HMGB1 mRNA were examined by PCR and the activity of diamine oxidase (DAO) were determined by ELISA.Results Compared with group S,the expression of HMGB1 mRNA was increased and the activity of DAO decreased in group Sep and groups L1～3 at 24 and 48 h after CLP ( P ＜ 0.05 ).Compared with group Sep,the expression of HMGB1 mRNA was decreased and the activity of DAO increased in a dose-dependent manner in groups L1～3 ( P ＜0.05),The injury of pathology of small intestine was slighter in groups L1～3 than in group Sep.Conclusion Lidocaine can reduce samll intestine injury through inhibiting HMGB1 expression in septic rats by a dose-dependent manner.

Objective To examine the levels of fractional exhaled nitric oxide in bronchial asthma and the cor-relation between fractional exhaled nitric oxide and pulmonary function tests as well as the value of pulmonary function tests in curing the children with bronchial asthma. Methods Seventy - two asthmatic children came from Guiyang Ma-ternal and Child Health Hospital were studied. They were divided into 2 groups:mild - to - moderate asthma group(36 cases)and serious asthma group(36 cases). Fractional exhaled nitric oxide level and the lung function were measured and the correlation analysis was analyzed by SPSS 13. 0 software. Results The fractional exhaled nitric oxide concen-tration of the serious asthma group was significantly higher than that of the mild - to - moderate asthma group[(47 ± 26)ppb vs(29 ± 15)ppb,t = 3. 543,P ﹤ 0. 01],1 - second rate,maximum mid - expiratory flow(MMEF),and peak expiratory flow(PEF)in mild - to - moderate asthma group were significantly higher than those in serious asthma group[(99. 74 ± 9. 81)% vs(87. 17 ± 13. 30)% ,(87. 12 ± 35. 61)% vs(68. 90 ± 30. 88)% ,(99. 97 ± 10. 83)% vs (94. 31 ± 17. 69)% ](P ﹤ 0. 01,0. 05,0. 05). No correlations were found between fractional exhaled nitric oxide level and the lung function parameters,including forced expiratory volume in 1s(FEV1),1 - second rate,MMEF and PEF in the mild - to - moderate asthma group(r = 0. 073,0. 087,0. 242,0. 326,all P ﹥ 0. 05). However the level of fractional exhaled nitric oxide was negatively correlated with the lung function parameters,including FEV1,FEV1 / FVC,MMEF and PEF in serious asthma group(r = - 0. 354,- 0. 573,- 0. 478,- 0. 471,P ﹤ 0. 05,0. 001,0. 005,0. 005),respec-tively. Conclusions Monitoring of fractional exhaled nitric oxide and the lung function parameters can effectively indi-cate the state of illness of asthmatic child,which plays a key role in the treatment.

Objective To investigate the changes of thyroid hormones in patients with peripartum car-diomyopathy. Methods Seventy-two patients with peripartum cardiomyopathy were consecutively enrolled and 72 healthy women who had normal delivery were regarded as control. Among all the subjects , free triiodothyronine (FT3), free thyroxine (FT4) and thyroid-stimulating hormone (TSH) in the serum and high sensitive C-reactive protein (hs-CRP), interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in the plasma were measured. Results The levels of serum FT3 and FT4 in peripartum cardiomyopathy group were significantly higher than those in control group (P < 0.05). And the level of serum TSH in peripartum cardiomyopathy group was significantly low-er than that in control group (P < 0.05). There were 24 cases with hyperthyroidism in peripartum cardiomyopa-thy group while no patient in the control group had hyperthyroidism (P < 0.05). The levels of plasma hs-CRP and IFN-γ in peripartum cardiomyopathy group were significantly higher than those in the control group (P <0.05). And the level of IL-4 in peripartum cardiomyopathy group was significantly lower than that in the control group (P < 0.05). Conclusions Serum thyroid hormones elevated in patients with peripartum cardiomyopathy and its mechanism might be related to abnormal immune reaction.

Objective To investigate the effect of vasoactive intestinal peptide (VIP) and methylprednisolone (MP) on Toll-like receptor (TLR)2/4 mRNA expression in endotoxin (lipopolysaccharide,LPS) induced shock.Methods Ninety Sprague-Dawley rats were randomly divided into LPS group (n =20),LPS + VIP group (n =20),LPS + MP group (n =20),LPS + VIP + MP group (n =20) and control group (n =10).LPS group injected intravenously LPS (E Coli O55B5) 10 mg/kg.LPS + VIP group,LPS + MP group and LPS + VIP + MP group were injected intravenously VIP 5 nmol/kg,MP 3 mg/kg and VIP 5 nmoL/kg + MP 3 mg/kg after LPS 10 mg/kg injection.The control group injected normal saline intravenously instead of LPS.The rats were sacrificed at 6 h and 24 h after injection and the intestine samples were collected.Pathological changes of the intestine were observed by microscopy.RT-PCR was used to detect the intestinal TLR2 mRNA and TLR4 mRNA expressions.Results Intestinal mucosa showed edema or necrotic change with structure of the microvilli disappeared after LPS injection.The inestinal lesions in VIP,MP and VIP + MP groups were milder than LPS group.At 6 h after LPS injection,TLR2 mRNA and TLR4 mRNA expressions were significantly up-regulated in LPS group,LPS + VIP group,LPS + MP group and LPS + VIP + MP group (TLR2 mRNA:1.14 ±0.38,1.17 ±0.42,1.16 ±0.41,0.92 ± 0.29;TLR4 mRNA 1.21 ±0.18,1.04 ± 0.38,1.11 ± 0.34,1.01 ± 0.20) compared with the control group (0.32 ± 0.20,0.24 ± 0.17) (P ＜ 0.01).But there was no significant difference between LPS group,LPS + VIP group,LPS + MP group and LPS + VIP + MP group (P ＞ 0.05).At 24 h after LPS injection,TLR2 mRNA and TLR4 mRNA expressions in LPS + VIP group,LPS + MP group and LPS + VIP + MP group (TLR2 mRNA:0.63 ± 0.12,0.59 ± 0.13,0.52 ±0.19;TLR4 mRNA 0.67 ±0.09,0.64 ±0.09,0.51 ±0.13) were significantly lower than LPS group (1.04 ± 0.38,0.82 ±0.18) (P ＜0.01) (P ＜0.05).Conclusion VIP and/or MP can mitigate intestinal injury induced by LPS shock.The gastrointestinal protection of VIP and glucocorticoids were related to downregulation signaling TLR2 mRNA and TLR4 mRNA expression.But VIP/MP and VIP + MP have no significant effect on expression of intestinal TLR2/4 mRNA until 24 h after LPS shock.

The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL^-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL^-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.

OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To explore the clinical features and diagnostic methods of tuberculosis infection in PICU,and improve the understanding of tuberculosis.Methods We analysed the clinical features and diagnostic methods of severe tubercle bacillus infectious diseases in PICU from Jan 2009 to Dec 2012.Severity of disease was graded by pediatric critical illness score.The diagnosis of the pulmonary tuberculosis was in accord with the diagnostic criteria of paediatric pulmonary tuberculosis established by Chinese Medical Association paediatrics branch.And the diagnosis of tuberculosis meningitis and tuberculosis peritonitis based on the clinical physical examination,laboratory examination and pathologic finding.Results Among 22 cases enrolled in this study,totally 16 cases were pulmonary tuberculosis,6 cases were extrapulmonary tuberculosis,and 3 cases were tuberculosis meningitis.The clinical feature of severe tuberculosis infection in PICU was accompanying with one or multiple organ dysfunction besides tuberculosis infection symptom,among them,respiratory dysfunction occurred in 16 cases,cardiovascular dysfunction was observed in 2 cases,and central nervous system dysfunction was found in 3 cases,even 1 patient experienced cardiovascular system dysfunction,respiratory disorder as well as gastrointestinal system dysfunction simultaneously.Sixteen cases of pulmonary tubercle bacillus infection manifested respiratory failure besides fever,cough,shortness of breath and tuberculosis toxicosis symptom,2 cases of them developed into acute respiratory distress syndrome,8 cases needed mechanical ventilation.Two cases of pericardial effusion presented cardiac tamponade.The level of adenosine deaminase elevated in 12 cases,and the positive result of enzyme-linked immunospot assay for tubercle bacillius was observed in 14 cases.Conclusion It is very important to be aware of that severe tubercle bacillus infection exist in critically ill patients admitted in PICU,measuring the level of adenosine deaminase and taking enzyme-linked immunospot assay for tubercle bacillius test are important accessory examination for tuberculosis diagnosis in children.

ObjectiveTo investigate the effects of sleep deprivation on expressions of Mir-132,mir-134 in the different regions of rat brain.MethodsAll the male SD rats were divided into control group ( normal sleep group),sleep deprivation (SD).The modified multiple platform method (MMPM) was used to establish sleep deprivation model.Mir-132,mir-134 level was detected by real time PCR.ResultsMir-132 were significantly increased in SD groups in hippocampus compared with the control groups ( 51.87 ± 8.13 vs 67.25 ± 7.59 ) (P ＜0.01 ).Mir-134 were significantly decreased in SD groups compared with the control groups( 1.82 ±0.15 vs 1.45± 0.12 )(P ＜ 0.01 ).There were no statistically significant differences in cortex and thalamus (P ＞ 0.05 ).Cortex mir-132 level in SD group and control group was 1.57 ±0.10,1.48 ±0.11 respectively,and it was 1.37 ±0.09,1.36 ±0.11 in thalamus;Cortex mir-134 level in SD group and control group was 98.26 ± 5.17,100.80 ±4.15respectively,and it was 97.56 ± 6.28,91.01 ± 4.07 in thalamus.ConclusionThe upregulation of mir-132 and downregulation of mir-134 implies that two miRNAs did opposite actions in the processes of sleep deprivation.This findings indicate that hippocampus mir-132,mir-134 levels in the SD rat may reflect associated depressive patho-physiological processes.

Objective To study the changes of P-selectin and E-selectin in pediatric patients with critical illness ,and analyze their relationship with the severity and prognosis of diseases.Methods Forprospective study,42 critically ill patients admitted in pediatric intensive care unit (PICU ) from September,2012 to March,2013 as critically ill group were enrolled,and blood specimens were collected with 24 hours after admission.Another 42 cases blood samples were collected from children's physical examination as control group.The severity of the critically ill patients were evaluated by Pediatric Critical illness Score (PICS)and Pediatric risk of score mortality (PRISM)-III.The levels of serum P-selectin and serum E-selectin were measured by double antibody sandwich enzyme-linked immunoassay (ABC-ELISA). Results P-selectin and E-selectin in control group children and critically ill patients group were (37.23 ± 8.99)ng/mL,(36.24 ±17.82)ng/mL,and (107.24 ±35.53)ng/mL,(114.93 ±40.17)ng/mL, respectively.There were statistical differences between two groups (P=0.000).The levels of P-selectin and E-selectin in acute phase were higher than that of levels in recovery phase in critically ill group (P =0.000).Negative correlation was observed between P-selectin concentration and the PCIS score (r =-0.673,P=0.000),as well as E-selectin (r=-0.548,P=0.000).P-selectin level and E-selectin level based upon PRISMⅢ≥10 group were significantly higher than they in PRISMⅢ <10 group (P=0.003,P=0.014).In critically ill children,the differences in P-selectin,E-selectin were significant higher in death patients (P=0.003;P =0.000).Compared with the non-sepsis illness group,the level of P-selectin and E-selectin in the severe sepsis patients were significantly higher (P =0.04,P =0.025 ). Conclusions The levels of P-selectin and E-selectin are closely related to the severity and prognosis in critically ill children.Measuring the level of P-selectin and E-selectin could be used as a judegment the severity and to understand pathological physiological process.