Objective To investigate the changes and diagnostic significance in plasma undercarboxylated osteocalcin in children with Kawasaki disease (KD),especially with coronary artery lesions (CALs).Methods The data of 36 KD children were collected,who were inpatients at Department of Cardiovascular and Rheumatology,Shanxi Province Children's Hospital from January 2015 to December 2015,including 20 boys and 16 girls,aged (2.3 ± 1.1)years old.According to the course of the disease,KD children were divided into an acute stage group and a subacute stage group.Based on the echocardiography findings,KD children were subdivided into CALs group and no coronary artery lesions (NCALs) group.Twenty-five healthy children from the physical examination during the same period were selected as the healthy control group,13 boys and 12 girls,aged (2.6-± 1.0) years old.Plasma undercarboxylated osteocalcin level was measured by double antibody sandwich enzyme-linked immunosorbent method.Sigrnaplot 12.5software was used to analyze the data statistically,and the receiver-operating characteristic (ROC) curve was used to evaluate the diagnostic effect of plasma undercarboxylated osteocalcin in KD with CALs.Results The levels of plasma undercarboxylated osteocalcin in the healthy control group,the acute stage group and the subacute stage group were (16.4 ± 1.6) μg/L,(14.2 ± 1.6) μg/L,(14.3-± 1.7) μg/L,respectively.Compared with the healthy control group,the plasma undercarboxylated osteocalcin level in the acute stage and the subacute stage were significantly lower,the differences were statistically significant (q =6.088,5.687,all P ＜ 0.01).But there was no difference of plasma undercarboxylated osteocalcin level between the acute stage group and the subacute stage group (q =0.466,P ＞ 0.05).The levels of plasma undercarboxylated osteocalcin in acute stage group with CALs and acute stage group with NCALs were (12.9 ± 1.2) μg/L,(15.0 ± 1.4) μg/L.Compared with healthy control group,the plasma undercarboxylated osteocalcin levels of children with CALs and with NCALs were obviously decreased,the differences were statistically significant (q =8.711,3.891,all P ＜ 0.01).There was a statistical difference in plasma undercarboxylated osteocalcin level between the acute stage with CALs and the acute stage with NCALs (q =5.171,P ＜ 0.01).The plasma undercarboxylated osteocalcin levels of KD children with CALs in the subacute stage was (13.0-± 1.3) μg/L.Compared with acute stage,there was no statistical difference (t =0.257,P ＞ 0.05).There was a sensitivity of 79％,specificity of 82％,positive predictive value of 88％ and negative predictive value of 70％ for the 15.7 μg/L undercarboxylated osteocalcin for diagnosing KD.There was a sensitivity of 83％,specificity of 88％,positive predictive value of 83％ and negative predictive value of 88％ for the 13.7 μg/L undercarboxylated osteocalcin for diagnosing KD with CALs.Conclusions Osteocalcin is related to the pathogenesis and development of KD.Plasma undercarboxylated osteocalcin contributes to the diagnosis of KD with CALs.

Objective To develop a novel automatic water system for ventilator humidifier.Methods By hanging the infusion apparatus high,sustained water was filled by driving force of gravity.Liquid level in the thong of the infusion apparatus was detected by the sensor,and the closing and opening of the closing clip was controlled by solenoid valve.Results The closing clip opened when the liquid level inside the ventilator humidifier fell below the lowest setting value and purified water in the infusion bottle automatically flowed to the humidifier.When the liquid level reached the highest setting level,the closing clip was automatically closed.Conclusions The developed automatic water system for ventilator humidifier is effective,convenient,inexpensive,and realized a sustained,relatively steady and a small amount of water process.Meanwhile,humidifier water is stable,heating is uniform,and gas temperature is relatively constant,which can be applied in clinical use.

Objective To evaluate the effect of total alkaloids of Sophora alopecuroides L on the nervous behavior and the expression of neurotransmitters in rats. Methods 48 male SD rats were randomly divided into 4 groups:blank group,total alkaloids of Sophora alopecuroides L treatment with 4 mg·kg-1 ·d-1 ( low dose group) ,8 mg·kg-1 ·d-1( medium dose group) and 16 mg·kg-1 ·d-1( high dose group) groups. After successive intragastric administration for 30 days,the locomotor activity was applied to test the nervous behavior and emotional state of rats in each group. After behavioral tests were finished,the contents of trypto-phan (Trp),5-hydroxytryptophan (5-HTP),5-serotonin (5-HT),5-hydroxyindoleacetic acid (5-HIAA), norepinephrine (NE),epinephrine (E) and dopamine (DA) were detected by ELISA in serum and brain. Results In the experiment of locomotor activity,compared with blank group ((95.33±12.75) times),the numbers of horizontal movement of Sophora alopecuroides L in medium and high dose group ( ( 61. 64 ± 5.91),(64.62±5.79)times both P<0.05) were significantly reduced (P<0.05, P<0.01).Compared with the blank group ((4.33±0.66) times),the numbers of vertical motion of Sophora alopecuroides L in medium and high dose group ((3.05±0.09)times,(3.77±0.69)times) were significantly reduced (P<0.05, P<0.01). Compared with blank group ((2.25±0.39) number),the granule number in high dose group ((3.38±0.40) number) were significantly increased (P<0.05, P<0.01),and emotional stress degree increased. Meanwhile,compared with blank group,the content detection of 5-HT,5-HIAA in different dose groups of the total alkaloids of Sophora alopecuroides L was improved obviously(P<0.01),but there was no significant effect on the content of Trp,5-HTP,NE,E,DA(P>0.05). Correlation analysis indicated that the degree of au-tonomic activity in rats with the content of 5-HT,5-HIAA and DA in serum was negatively correlated (P<0.05, P<0.01) ,the degree of emotional stress and the content of 5-HT,5-HIAA in brain was negatively cor-related (P<0.05, P<0.01) . Conclusion The total alkaloids of Sophora alopecuroides L can reduce the ac-tivity of rats and increase the degree of emotional stress. And the mechanism may be correlated with the in-creasing level of 5-HT and 5-HIAA in serum and brain.

Aim To study the insulinotropic effects of Efaroxan and the underlying mechanism in rat βcells. Methods Pancreatic islets were isolated by college-nase p digestion.Radioimmunoassay was used to meas-ure insulin secretion and cAMP level in rat pancreatic islets.Results Efaroxan only potentiated insulin se-cretion at high glucose concentrations(8.3,1 1 .1 mmol ·L -1 )but not at low glucose concentrations.KU1 4R,an antagonist of Efaroxan,remarkably inhibited Efarox-an-potentiated insulin secretion;and similarly,KU1 4R significantly inhibited forskolin-induced and IBMX-in-duced insulin secretion.cAMP measurement showed that forskolin and IBMX significantly increased cAMP levels,but Efaroxan and KU1 4R had no effects on cAMP content in pancreatic islets.Conclusion The mechanism of Efaroxan-potentiated insulin secretion is related to downstream of cAMP signaling pathway, KU1 4R antagonized the downstream of cAMP signaling leading to its inhibitory effects on Efaroxan,forskolin and IBMX-induced insulin secretion.

Aim To investigate the effects of 3 ,5 ,2 ’ , 4’-tetrahydroxychalcone (P40) on urate excretion, as well as mRNA and protein expressions of renal URAT1 and GLUT9 in hyperuricemic mice. Methods Sixty Kunming mice were randomly divided into six groups:normal control group, hyperuricemic group ( model group), P40 2. 0, 4. 0, 8. 0 mg·kg-1 groups and positive control group. All drugs were administered in-tragastrically to mice for 5 doses. Hyperuricemic mice were induced by intraperitoneal injection of uric acid (0. 15 g·kg-1 body weight) for 3 times. The urate levels were assayed with the phosphotungstic acid method. The mRNA and protein expressions of GLUT9 and URAT1 were determined by RT-PCR and Western blot. Results P40 at a dose of 4. 0 and 8. 0 mg · kg-1 significantly reduced the serum urate levels in a dose-dependent manner, when compared with untreat-ed hyperuricemic mice ( P<0. 05 or 0. 01 ) . The he-patic urate contents decreased in untreated-and treated-hyperuricemic mice as compared with normal mice ( P<0. 01 ) . Furthermore, P40 had no influence on the renal URAT1 mRNA and protein expression levels, while it could down-regulate renal GLUT9 protein ex-pression but not mRNA expression in hyperuricemic mice. Conclusion P40 possesses potent uricosuric effects associated with urate reabsorption by down-regu-lating the protein expression of GLUT9 in kidney.

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated specklike protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

OBJECTIVE:To study the effects of butin in Vernohia anthelmintica(VW)on proliferation of human immortal ke-ratinocyte cell strain HaCaT and cell secretory factors,and explore the mechanism of butin in VW in the treatment of vitiligo. METHODS:MTT method was used to determine the survival rate of HaCaT cells cultured by 0 (blank control),0.1,0.5,1.0, 5.0,10.0 μg/mL of butin for 48 h. Enzyme-linked immunosorbent assay was used to determine the contents of cell secretory factors as endothelin 1 (ET-1),ET-3,melanocyte stimulating hormone (MSH),stem cell factor (SCF),basic fibroblast growth factor (bFGF)in culture medium after HaCaT cells were cultured by 0.5,1.0,5.0 μg/mL of butin for 48 h. RESULTS:Compared with blank control,cell survival rate was increased to varying degrees after cultured by 0.1-5.0μg/mL of butin for 48 h,while decreased after cultured by 10.0 μg/mL of butin. Contents of ET-1,SCF,bFGF in culture medium were significantly increased after cultured by 0.5,1.0,5.0μg/mL of butin for 48 h(P<0.01);and contents of ET-3,MSH in culture medium were significantly increased af-ter cultured by 1.0,5.0 μg/mL of butin for 48 h(P<0.01). CONCLUSIONS:Butin can promote the proliferation of HaCaT cells, the mechanism may be associated with promoting the secretion of cell secretory factors of ET-1,ET-3,MSH,SCF,bFGF.

Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P<0.01), there were significant difference between cervical squamous cancer and HSIL, or between HSIL and LSIL(all P<0.05), while there were not significant difference between LSIL and chronic cervicitis(P>0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P<0.05), but not with age, clinical stage, lymph node metastasis, and serum squamous cell carcinom antigen(SCC-Ag)levels (all P>0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.

Purpose To investigate the diagnostic value of prenatal MRI in fetal intracranial hemorrhage (FICH). Materials and Methods The imaging and clinical data of 41 cases of FICH accepting MRI diagnosis were retrospectively analyzed. The hemorrhage location, signal characteristics and the associated intracranial abnormalities were observed. The pregnancy outcomes and clinical data after birth were followed up. The correlation between periventricular hemorrhage/intraventricular hemorrhage (PVH/IVH) classification and clinical outcomes was analyzed by combining prenatal risk factors. Results Forty-one cases of FICH were evaluated. There were 23 cases of multifocal cerebral hemorrhage and 18 cases of single focal hemorrhage. According to the bleeding site, the 41 cases were classified into PVH/IVH (33 cases), cerebral hemispheres near cortex hemorrhage (3 cases), cerebellar hemorrhage (2 cases), subdural hemorrhage (2 cases) and subarachnoid hemorrhage (1 case). Most of the FICH cases were in subacute period (36/41) and a few were in chronic period or forming encephalomalacia (5/41). The associated changes included ventriculomegaly, vascular malformation, agenesis of corpus callosum, agenesis of vermis, etc. Follow-up results showed that there were 25 cases of labor induction (autopsy after labor induction was performed in 3 cases), 16 cases were followed-up after birth. Among the 16 newborn, there was 1 case of PVH/IVH grade II fetus showing left ear hearing loss after birth, 1 case of grade II fetus showed dyskinesia within one year after birth, and 1 case of grade IV fetus showed unilateral limb movement disorder. The other 13 cases had no obvious neurological sequelae. Spearman correlation test of ranked data indicated that PVH/IVH classification was moderately correlated with birth outcome (r=0.689, P<0.05). Conclusion Prenatal MRI can evaluate the type and severity of fetal intracranial hemorrhage, and provide references for clinical diagnosis and treatment.

OBJECTIVETo investigate the mortality rates of hepatocellular carcinoma (HCC) in Nantong,China from 1999 to 2011, in order to uncover dynamic trends and provide reasoned advice on intervention strategies to decrease HCC incidence and mortality in Nantong in the future.

METHODSVersions 10 and 9 of the WHO International Classification of Diseases (ICD-10 and ICD-9) were used to determine the number of HCC deaths in Nantong,China for the study's range of years. Thex2 test was applied to compare the HCC mortality rates according to sex and age. The Grey system GM(1,1) model was used to predict the next-5-year HCC mortality for Nantong.

RESULTSAnalysis of the standardized mortality in Nantong showed a slight decreasing trend from 1999 to 2011 (x2=57 545.98, P less than 0.001),with males showing a steeper decrease than females. The total mortality of HCC during these years was 53.41 per 100,000 people,with mortality among males being significantly higher than that among females (80.81 per 100,000 people vs. 26.94 per 100,000 people; x2=13 625.42, P less than 0.001). In general, HCC mortality increased with increase in age (general trend:x2=57 545.98, P less than 0.001; male trend: x2=39 878.8, P less than 0.001; female trend: x2=20 105.3, P less than 0.001). However,HCC mortality increased significantly in women after the age of 40 and in men after the age of 35. The GM(1,1) equation was: Yt=-1265.28e(-0.0375t)+1315.5, which predicted that the HCC mortality will decrease to 25.56 per 100,000 people in 2016.

CONCLUSIONAlthough HCC mortality generally decreased from 1999 to 2011, the rate remained high. Public health intervention strategies may be more effective if they focus on males over the age of 35 and females over the age of 40.

Carcinoma, Hepatocellular ; mortality ; China ; epidemiology ; Female ; Humans ; Incidence ; Liver Neoplasms ; mortality ; Male