Objective Observe the changes of cytokine and immunoglobulin in children with Hand-foot-and-mouth disease (HFMD) infected by Enterovirus 71 (EV71),to explore the significance of inflammation and immunological mechanisms.Methods A total of 65 cases of children with HFMD by EV71 were taken as the observation group,and 65 cases of healthy children were as the control group in June 2015-August 2016.The serum levels of IL-6,IL-8,TNF-α were detected by enzyme-linked immunosorbent assay (ELISA),and the levels of IgA,IgG,IgM were detected by immune scattering turbidimetric.Results Compared with control group,the levels of IL-6,IL-8 and TNF-α in observation group were significantly increased (t=7.94,6.87,9.56,P ＜ 0.01),the levels of IgA,IgG and IgM also increased,differences were statistically significant (t =6.71,7.96,8.39,P ＜ 0.01).Conclusion The inflammatory cytokines and immune imbalance in the development of HFMD play an important role,observe the dynamic changes of IL-6,IL-8,TNF-α,IgA,IgG and IgM levels has certain clinical application value in preventing and judging the state of HFMD.

Objective To establish the newborn rhesus monkey model of hemolytic hyperbilirnbinemia and provide an experimental basic model for research of hyperbilirubinemia.Methods Sixteen 3-day old newborn rhesus monkeys were divided into experimental group and control group,with 8 newborn rhesus monkeys in each group.Eight newborn rhesus monkeys in experimental group were treated with intravenous injection of l0 g/L phenylhydrazine hydrochloride (50 mg/kg) to establish model of homolytic hyperbilirubinemia.The newborn rhesus monkeys in control group were treated with intravenous injection of 9 g/L saline at the same time.Twenty-four hours and 48 hours after the experimental treatment,the bilirubin in blood was detected to evaluate the models,and the clinical manifestations of newborn rhesus monkeys with hyperbilirubinemia were recorded by using monitoring equipment.The brain slices were made to evaluate the model in 1 dead monkeys of experimental group.Results The newborn rhesus monkey of experimental group showed obvious skin,sclera jaundice and hemoglobinuria.The serum total bilirubin [(252.76 ± 63.42) μmol/L],unconjugated bilirubin[(165.85 ±44.93) pmol/L] and conjugated bilirubin [(87.16 ±21.22) μmol/L] in the experimental group were significantly higher than those [(20.62 ± 5.72) μmol/L,(7.93 ± 2.31) μmol/L,(12.51 ± 3.53) μmol/L] in the control group,and the differences were statistically significant (t =14.581,13.881,14.040,all P ＜ 0.01).The level of hemoglobin [(47.18 ± 10.09) μmol/L] in the experimental group was significantly lower than that of the control group [(136.85 ± 13.48) μmol/L],and the difference was statistically significant (t =-21.308,P ＜ 0.01).The results of pathological showed brain edema,rupture and eosinophilic and bilirubin deposition in the basal nuclei,and necrosis appeared in some severe parts.And there were different degrees of retardation and coordination disorders in the experimental group(s) newborn rhesus monkeys,but gradually returned to normal in 4 months later.Conclusion Intravenous injection of phenylhydrazine hydrochloride can be used to produce newborn rhesus monkey models of hemolytic hyperbilirubinemia.

One strain of PRRSV was isolated from tissue of piglets died of obvious respiratory syndrome and high fever.It was identified as a strain of American type PRRSV by serological test and gene sequencing.According to American type PRRSV VR-2332,we designed 3 pairs of primers to aim directly at N gene,GP5 gene and variation sequence of NSP2.The results of sequence analysis indicated a discontinuous deletion of 30 amino acids in nonstructural protein 2 (NSP2).N and GP5 gene is conservative relatively.

Objective To investigate the clinical applications of the chain locking-type tension bands. Methods89 cases with patellar bone fractures,27 olecranal fractures patients,18 patients with fractures of surgical neck of humerus,16 patients with dislocation of the acromioclavicular joints and 12 patients with clavicular lateral fractures were treated with chain locking-type tension bands. ResultsTheResultsshowed that all patients wound were postoperative first intention.The healing time of the fractures were 6 ～ 18 months(average 10 months).The Kirschner's pins lapping,steel wire breaks and tension bands out of control were not found. ConclusionCompared with the traditional tension bands,the chain locking-type tension bands had stronger stability,stress distribution more even and less complications.

Diffuse intrinsic pontine glioma( DIPG)is a highly invasive tumor located in the pons (middle)of the brain stem. They are usually diagnosed during childhood and account for 10% -15% of primary brain tumors in children. DIPG has a very poor prognosis. Fewer than 10% of DIPG patients survive more than 2 years after diagnosis. The imaging manifestations of DIPG are typical,and biopsy is only performed in atypi-cal cases. The tissue specimens of newly diagnosed DIPG are very few and limit its molecular biological research. Recent advances in surgical and molecular-analytic techniques have increased the safety of biopsy which has already been used in many clinical trials step by step. The research of DIPG′s molecular pathogenesis and treatment is sure to achieve new breakthroughs.

Objective To study the effect of endotoxin tolerance (ETT) on chemokine receptor 7 (CXCR7) in the liver tissue of rats with acute liver failure (ALF).Methods SD male rats were randomly divided into three groups:normal group,ALF group and ETT group.The rats in the ETT group and ALF group were injected with lipopolysacharide (LPS) 0.1 mg/kg or saline respectively,one time / day for 5 days.At 24 hours after the 5th-day injection,all rats were injected with D-GalN 800 mg/kg and LPS 8μg/rat.Blood sample and liver tissue were collected on 2,6,12,24 and 48 hours after injection.The gene expressions of CXCR7 in the liver were measured by RT-PCR,and the protein expressions of CXCR7 were determined by Western Blot.The data analysis was performed by LSD,Dunnett's t test.Results The histological damage in the liver tissue was significantly mider in ETT group compared to ALF group.The gene expressions of CXCR7 were significantly milder in ETT group compared to ALF group (2 h:F =29.222,6 h:F=166.892,12 h:F=38.975,24h:F=34.603,48 h:F=18.929,allP＜0.01),but still severer than that of normal group.The CXCR7 protein expression in ALF group and ETT group peaked at 24 hours,but the expression of CXCR7 in ETT group was lower compared with that in in ALF group (2h:F=11.155,6 h:F=42.553,12h:F=17.082,all P＜0.01; 24 h:F=7.242,P＜0.05).Conclusions During the process of endotoxin tolerance,LPS pretreatment and D-GalN can decrease the liver injury,down-regulate the expressions of CXCR7mRNA and CXCR7.This suggests that CXCR7 may play an important role in the ETT.

Objective To explore the dynamic expressions of interleukin-1 (IL-1)receptor associated kinase (IRAK)-M and IRAK-4 in rats with or without endotoxin tolerance (ETT)in acute liver failure (ALF).Methods Sixty-six male SD rats were divided into three groups:ALF group,ETT group and control group.The rats in ETT group received daily lipopolysaccharide (LPS)intraperitoneal injection for 5 days,while the rats in ALF group received daily injection with same volume of 0.75% NaCl solution.Both ETT group and ALF group received intraperitoneal injection with D-galactosamine (D-GalN)and LPS at 24 hours after the 5th injection of LPS or NaCl solution.At 2,6,12,24 and 48 h after D-GalN and LPS injection,the serum levels of tumor necrosis factor-α (TNF-α)and interleukin-6 (IL-6)were detected by enzyme-linked immunosorbent assay (ELISA).The liver pathologic changes were observed with HE staining by microscope.The mRNA expressions of IRAK-4,IRAK-M and NF-κB (p65)were detected by reverse transcriptase polymerase chain reaction (RT-PCR).The pairwise comparison between groups was done by lease significant difference (LSD)and Dunnet's t test.Results The liver pathologic changes in ETT group were much milder than those of ALF group.In ALF group,IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24and 48 h after D-GalN and LPS challenge were 0.711 ±0.074,0.904±0.118,1.012 ±0.098,1.534±0.279 and 1.451±0.290,respectively,while the IRAK-M mRNA/β-actin absorbance ratios were 0.496±0.018,0.516±0.089,0.503±0.023,0.503±0.057 and 0.469±0.142,respectively.In ETT group,the IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24 and 48 h after D-GalN and LPS challenge were 0.619±0.083,0.587±0.033,0.623±0.034,0.720±0.044 and 0.654±0.041,respectively,while the IRA'K-M mRNA/β-actin absorbance ratios were 0.929 ± 0.064,1.111±0.138,1.113±0.027,1.891±0.315 and 1.710±0.303,respectively.The IRAK-M mRNA expression level was significantly increased and IRAK-4 mRNA expression level was relatively decreased in ETT group compared to ALF group.The differences were all statistically significant at 2,6,12,24 and 48 h after D-GalN and LPS challenge (F= 17.305,54.921,121.031,67.607,55.279,respectively; F=19.506,43.777,110.823,302.681,202.822,respectively; F=172.003,59.519,987.055,68.463,96.601,respectively; all P＜0.05).Conclusion LPS pretreatment may induce ETT in rat model by downregulating the expression of IRAK-4 and upregulating the expression of IRAK-M.

Objective To investigate the mechanism of endotoxin tolerance (ETT) through observing the expression of OX40 in liver tissues of ETT rats.Methods SD male rats were randomly divided into three groups:normal group (n=6),acute liver failure (ALF) group (n=24) and ETT group (n=24).Lipopolysacharide (LPS) 0.1 mg/kg (ETT groups) or 0.9 ％NaC1 (ALF groups) was administered by five consecutive intraperitoneal injections at 24 h intervals,and at the sixth day,all animals were treated with intraperitoneal injections of D-galactosamine (D-GalN) 800 mg/kg and LPS 8 μg/rat.Blood and liver tissue were collected at 6,12,24 and 48 hours after the injection of D-GalN/LPS.The gene expression of OX40 in the liver was measured by reverse transcription-polymerase chain reaction (RT-PCR).The protein expression of OX40 was estimated by Western blot.The tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels were determined by enzyme-linked immunosorbent assay (ELISA).The data analysis was performed by one-way ANOVA,lease significant difference (LSD) and Dunnett's T3 test.Results The expressions of TNF α and IL-6 were both significantly lower in ETT group compared to ALF group,but still higher than that of control group.The gene expressions of OX40 peaked at 12 hours and decreased gradually in ALF group.The gene expressions of OX40 were significantly lower in ETT group compared to ALF group (6 h:F=5.027,24 h:F=5.539,48 h:F=5.011,all P＜0.05; 12 h:F=36.688; P＜0.01),but still higher than that of normal group.The tendency of OX40 protein expression in ALF group was peaked at 12 hours and decreased at 24 hours.In ETT group,the expression of OX40 was lower,and the difference between ETT group and ALF group had statistical significance (6 h:F=8.658,P＜0.05; 12 h:F=34.611,24 h:F=28.176,48 h:F=16.747; all P＜0.01).Conclusions The level of OX40 is increased in ALF group,while the expressions of OX40,TNF-α and IL-6 are lower in ETT group,which suggested that OX40 may play an important role in the process of ETT.