Objective To analyze the sandtray topics of the children with behavior problems according to the grounded theory method.Methods Collected and described children' s sandtray topics of different types of behavior problems,age and gender.And analyzed them base on a 3 levels coding list compiling according to the ground theory method.Results The frequency of “negative theme” (3.33 ± 1.25 ) was higher than “positive theme” (0.81 ± 1.25 ) with significant difference (P＜0.01) in feature level 1.Age factor had significant effect upon the frequency of “positive theme” in feature level 1 (P ＜ 0.05 ).And also had impact on “harmony”,“ development” in features level 2 (F =3.039,P ＜ 0.05 ).Gender factor,interaction of gender and age had significant effects on frequency of “negative theme” in feature level 1 (F =5.395,P ＜ 0.05 ; F =4.222,P ＜ 0.05 ).Conclusion The characteristics of the children' s sandtay with behavioral problems mainly are “negative theme”.And it shows different features in children with different gender or age.

Objective To investigate the value of nerve conduction studies (NCSs),F wave analysis,somatosensory evoked potential (SEP) and skin sympathetic response (SSR) in the early diagnosis of diabetic peripheral neuropathy (DPN).Methods A total of 110 patients with diabetes mellitus were recruited as the diabetic group and another 50 well-matched healthy volunteers as the normal controls.Sensory and motor NCSs of the median,ulnar,posterior tibial and common peroneal nerves were performed.F waves were recorded from the median and posterior tibial nerves.SEPs elicited by stimulation to nerves of both the upper and lower limbs as well as SSRs were measured,all in both the diabetic group and the normal controls.Results The total rate of nerve conduction abnormality was 74.5％ in the diabetic group,with sensory nerve conduction abnormalities more frequent and more severe among motor nerves in the extremities.The total rate of F wave abnormalities was 57.3％ in the diabetic group.The rate in patients with normal distal motor conduction in their median and posterior tibial nerves was 50.7％.The total SEP abnormality rate was 70.0％ with regard to the proximal peripheral nerve potentials in the diabetic group,but there was no obvious abnormality of the supraclavicular electrical potential in the upper limbs for those with normal sensory nerve conduction in the median nerve.The rate of occurrence of abnormality in the gluteus point potential in the lower limbs of those with normal posterior tibial sensory conduction was 62.5％.The total rate of SSR abnormalities was 80.0％ in the diabetic group but 72％ among those with normal nerve conduction in their extremities.Combining the NCS,SSR,SEP and F wave results,the total abnormality rate was 90.9％ in the diabetic group,which was much higher than with any single test used alone.Conclusion NCS is essential for diagnosing DPN.Early diagnosis of subclinical diabetic neuropathy will be significantly enhanced when nerve conduction,SSRs,SEPs and F waves are tested together.

Objective To study the differences of coagulation indices on the first day of birth in newborns with different gestational ages.Method Premature infants born in our hospital between January 2014 and December 2015 were enrolled in this study as the observational group,and they were divided into early preterm group,moderate preterm group,and late preterm group according to their gestational ages.Healthy full-term infants born during the same period were selected as the control group by 3:1 The clinicaldata and coagulation indices of the infants and their mothers in each group were compared.Result There were 44,50,71,and 52 cases in the early preterm,moderate preterm,late preterm,and control group,respectively.The prothrombin time (PT),activated partial thromboplastin time (APTT),and thrombin time (TT) of the premature infants in the early preterm group,moderate preterm group,and late preterm group were all longer than those of the control group [PT:(16.1 ±4.3) s,(16.8 ±4.9) s,(15.8 ±4.8) s,vs.(13.0±1.3)s;APTT:(88.3±38.1) s,(93.5±37.7) s,(91.0±32.3) s,vs.(66.0±17.8) s;TT:(25.4 ±4.6) s,(25.1 ±5.5) s,(25.0 ±3.3) s,vs.(24.0 ±3.3) s;all P＜0.05].The fibrinogen level of the premature newborns in three groups were all lower than that of the full-term infants in the control group [(1.11 ± 0.46) g/L,(1.12 ± 0.44) g/L,(1.12 ± 0.45) g/L vs.(1.28 ± 0.37) g/L,P ＜ 0.05].The differences of all the indices among the three groups of premature infants were all not statistically significant (P ＞ 0.05).The comparison of the coagulation indices of the mothers of the newborns from all four groups showed no significant differences (P ＞ 0.05).Conclusion Compared with full-term infants,preterm infants showed significantly poorer coagulation function on the first day of birth.However,there were no significant differences in coagulation indices among preterm infants of different gestational ages.

Objective To evaluate the effects of ulinastatin on renal ischemia-reperfusion injury in patients undergoing operation on aorta with deep hypothermic circulatory arrest (DHCA ) .Methods Thirty patients ,aged 30-50 yr ,of ASA physical status Ⅲ or Ⅳ (NYHA Ⅱ or Ⅲ) ,scheduled for elective operation on aorta with DHCA ,were randomly divided into 2 groups ( n=15 each) using a random number table :control group (group C ) and ulinastatin group (group U ) .In group U ,ulinastatin 20 000 U/kg was infused via the central vein at 500-1 000 U·kg-1 ·min-1 from the time immediately after tracheal intubation until 10 min before ascending aortic cross-clamping .In group C ,the equal volume of normal saline was infused instead of ulinastatin .At 5 min before the beginning of DHCA (T1 ) and 15 min after the end of DHCA (T2 ) ,blood samples were taken from the extracorporeal circulation for determination of polymorphonuclear leukocyte counts , and plasma levels of intercellular adhesion molecule-1 , tumor necrosis factor-α, iterleukin-6 (IL-6 ) IL-8 , IL-10 , malondialdehyde , myeloperoxidase ,atrial natriuretic peptide ,cystatin C ,and creatinine .Results The polymorphonuclear leukocyte counts and plasma levels of intercellular adhesion molecule-1 , tumor necrosis factor-α, IL-6 , IL-8 , malondialdehyde , myeloperoxidase , cystatin C , and creatinine were significantly lower , and the plasma concentrations of IL-10 and atrial natriuretic peptide were higher in group U than in group C ( P< 0.05 ) . Conclusion Ulinastatin can attenuate renal ischemia-reperfusion injury in patients undergoing operation on aorta with DHCA and inhibition of inflammatory responses is involved in the mechanism .

Objective To explore the synergetic effect of HBX protein and M2 macrophages in inflammatory microenvironment on invasion and metastasis of hepatocellular carcinoma cells.Methods Hep3B cells were infected with recombinant lentivirus carrying HBx gene,following co-culture with THP-1 original M2 macrophages.The cells were divided into six groups:two infected groups (Hep3B +and Hep3B + + M2),four non-infected groups (Hep3B-,Hep3B-+ LV5,Hep3B-+ M2,Hep3B-+LV5 + M2).Western blot (WB) was used to assess the expression changes of E-cadherin and N-cadherin,markers of epithelial-mesenchymal transition (EMT).The cellular location of EMT markers was observed by immunofluorescence confocal microscopy.Transwell assay was used to evaluate the invasion ability of Hep3B cells.Results HBX protein overexpressed in Hep3B cells by lentivirus infection.After 72 h co-culture with M2 macrophages,WB results showed that E-cadherin descreased significantly in Hep3B+ (0.42 ±0.11) when compared with Hep3B-(1.00 ±0.18) (t =4.762,P ＜0.05),while N-cadherin was significantly higher in Hep3B + (2.85 ± 0.44) than in Hep3B-(1.00 ± 0.17) (t =4.762,P ＜ 0.05).M2macrophages decreased E-cadherin expression in Hep3 B + + M2 (0.1 ± 0.13) compared with Hep3 B + (t =3.255,P ＜0.05),while N-cadherin expression increased in Hep3B+ + M2 (4.18 ± 0.52) (t=10.009,P ＜ 0.05).Non-Infected groups didn't change the markers of E-cadherin and N-cadherin.It was suggested that invasion ability of Hep3B increased by HBx overexpression.Conclusions HBX protein and M2 macrophages synergetically mediated the invasion and metastasis of hepatocellular carcinoma cells by EMT.

Objective To detect the expressions of osteopontin (OPN) and inerleukin-18 in patients with bullous pemphigoid (BP),and to analyze their relationship with clinical and laboratory indices.Methods Enzyme linked immunosorbent assay (ELISA) was performed to quantify the serum levels of osteopontin (OPN) and inerleukin (IL)-18 in 30 patients with BP and 30 health controls.Results The serum level of OPN was statistically higher in the patients than in the healthy controls ((8.29 ± 2.76) vs.(3.88 ± 1.41 ) ng/ml,P ＜ 0.01 ),and was positively correlated with the severity of BP (r =0.658,P ＜ 0.01 ) and with some laboratory indices in the patients.Increased serum IL-18 level was observed in patients complicated by cardiovascular diseases compared with those without cardiovascular diseases ((37.49 ± 6.43) vs.(31.10 ± 5.40) pg/ml,P ＜ 0.01).Moreover,the BP patients with diabetes,tumor,hepatic and renal impairment displayed an enhanced level of serum OPN and IL-18 than those without (all P ＜ 0.05).Conclusions OPN may be positively correlated with the severity of BP,while IL-18 may be involved in the development of complications of BP.

Objective To evaluate the effects of ulinastatin preconditioning on protamine-induced pulmonary injury in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).Methods Sixty NYHA class Ⅱ or Ⅲ and ASA physical status Ⅱ or Ⅲ patients of sexes,aged 21-59 yr,scheduled for elective cardiac valve replacement under CPB,were randomly divided into 3 groups (n =20 each):group control one protamine given via central vein (group C1) ; group control two protamine given via ascending aorta (group C2) ;group ulinastatin preconditioning (group U).Heparin was neutralized with protamine after termination of CPB.Ulinastatin 20 000 U/kg was infused via the central vein at a rate of 500-1000 U· kg-1 · min-1 starting from the time point after tracheal intubation until 10 min before cross-clamping of superior vena cava and inferior vena cava in group U.At 10 min after termination of CPB,protamine 4 mg/kg was infused over 8 min via the right internal jugular vein in groups C1 and U,or via the aortic root in group C2.Blood samples were obtained from the left atrium and right atrium at 5 min before neutralization of heparin with protamine (T1) and 15 min after neutralization of heparin with protamine (T2) for determination of polymorphonuclear leukocyte (PMN) and platelet (Plt) counts,and plasma concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-keto-PGF1α).Blood samples were obtained from the left atrium at T1 and T2 for determination of the levels of TNF-α,IL-1,IL-8,CD11b/CD18,C3a,C5a,and malondialdehyde (MDA) and superoxide dismutase (SOD) activity and for blood gas analysis.Alveolar-arterial oxygen gradiant (A-aDO2),respiratory index (RI) and oxygenation index (OI) were calculated.Pulmonary arterial pressure (PAP) was recorded.Results Plt and PMN counts in the blood obtained from the left atrium were significantly lower,and plasma TXB2 concentrations in the blood obtained from the left atrium were higher at T2 in group C1,and the plasma 6-keto-PGF1α concentrations and SOD activity in the blood obtained from the left atrium were higher at T2 in groups C2 and U than those in the blood obtained from the right atrium (P ＜0.05).Compared with group C1,Plt and PMN counts and plasma 6-keto-PGF1α concentrations were significantly increased,the levels of plasma TXB2,TXB2/6-keto-PGF1α,TNF-α,IL-1,IL-8,C3a,C5a and MDA were decreased,CD11b/CD18 expression was down-regulated,PAP,A-aDO2 and RI were decreased,and OI was increased at T2 in C2 and U groups (P ＜ 0.05).There were no significant differences in the parameters mentioned above between groups C2 and U (P ＞ 0.05).Conclusion Ulinastatin preconditioning can inhibit protamine-induced pulmonary injury in patients undergoing cardiac valve replacement under CPB,and the effect is similar to that of protamine administered via the aorta.

Objective To investigate the role of phosphatidylinositol 3-kinase (PI3K)/protein-serine-threonine kinases (Akt) signal pathway in ulinastatin postconditioning-induced attenuation of apoptosis in myocardial cells in patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Forty NYHA class and ASA physical status Ⅱ or Ⅲ patients of both sexes,aged 21-59 yr,scheduled for cardiac valve replacement with CPB,were randomly divided into 2 groups (n =20 each):normal saline control group (group C) and ulinastatin postconditioning group (group U).In group U,ulinastatin 10 000 U/kg was perfused via the aortic root at 4000-5000 U· kg-1 · min-1 starting from 5 min before aortic unclamping.In group C,the equal volume of normal saline was given instead of ulinastatin.Myocardial specimens were taken from the right auricle at 45 min after aortic unclamping for determination of the expression of Akt,phosphorylated Akt (p-Akt),cytochrome c,caspase-9,Bcl-2 and Bax,and cell apoptosis.Bcl-2/Bax ratio and apoptotic index were calculated.Results The expression of p-Akt and Bcl-2 and Bcl-2/Bax ratio were significantly higher,and the expression of cytochrome c,caspase-9 and Bax and apoptotic index were lower in group U than in group C (P ＜ 0.05).Conclusion Ulinastatin postconditioning attenuates apoptosis in myocardial cells in patients undergoing cardiac valve replacement with CPB through activating PI3K/Akt signal pathway.

BACKGROUND:Adipose-derived mesenchymal stem cells are gaining widespread interest in the Achil es tendon tissue engineering and regeneration, and an enabling environment (oxygen concentration) for cellinduction and differentiation is particularly necessary.
OBJECTIVE:To co-culture adipose-derived mesenchymal stem cells with primary tenocytes in standard culture condition (20%O 2 tension) and in an atmosphere of reduced oxygen (2%O 2 tension) in order to determine whether the two conditions differ in their effect on tenogenic differentiation.
METHODS:Tenocytes were isolated via serial expansion in culture from several Sprague-Dawley rats’ Achil es tendons. Adipose-derived mesenchymal stem cells were purchased. After one passage, adipose-derived mesenchymal stem cells were indirectly co-cultured with tenocytes in standard culture condition (20%O 2 tension) and in an atmosphere of reduced oxygen (2%O 2 tension). Col agen 1, col agen 3, Tenomodulin, Thrombospondin-4, Scleraxis levels were compared for each culture condition at 7, 14 and 21 days fol owing co-culturing. Immunofluorescence staining was performed to evaluate production of col agen 1 and Thrombospondin-4.
RESULTS AND CONCLUSION:Fol owing indirect co-culturing, hypoxic differentiated adipose-derived mesenchymal stem cells expressed higher levels of tendon-related genes and proteins than normoxic controls, which suggest that oxygen levels can significantly affect tenogenic differentiation, and hypoxia is advantageous for efficient differentiation of adipose-derived mesenchymal stem cells in vitro for tendon tissue engineering.

Objective To evaluate the role of Fas/FasL signaling pathway in ulinastatin postconditioning-induced attenuation of apoptosis in the myocardial cells of patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Forty patients of both sexes,aged 21-59 yr,of ASA physical status Ⅱ or Ⅲ (NYHA class Ⅱ or Ⅲ),scheduled for elective cardiac valve replacement with CPB,were randomly divided into 2 groups (n =20 each):control group (group C),and ulinastatin postconditioning group (group U).In group U,ulinastatin 10 000 U/kg was perfused via the aortic root at 4 000-5 000 U·kg-1 ·min-1 starting from 5 min before aortic unclamping.In group C,the equal volume of normal saline was infused instead of ulinastatin.Myocardial specimens were taken from the right auricle at 45 min after aortic unclamping for determination of Fas,Fas ligand (FasL),caspase-8,Bcl-2 and Bax expression and cell apoptosis.The ratio of Bcl-2 expression to/Bax expression (Bcl-2/Bax) and apoptotic index were calculated.Results Fas,FasL,caspase-8 and Bax expression and apoptotic index were significantly lower,and Bcl-2 expression and Bcl-2/Bax were higher in group U than in group C.Conclusion Ulinastatin postconditioning attenuates apoptosis in the myocardial cells through inhibiting Fas/FasL signaling pathway in the patients undergoing cardiac valve replacement with CPB.