Tocilizumab is a recombinant humanized monoclonal antibody against interleukin(IL)- 6 receptor (IL - 6R). It can prevent IL - 6 from binding to membrane - bound or soluble IL - 6R,thus blocking IL - 6 mediated signal transduction,and clinically alleviating inflammation and joint destruction in rheumatoid arthritis(RA). Both in the American and European Union,intravenous Tocilizumab has been approved for the treatment of both systemic juve-nile idiopathic arthritis(sJIA)and polyarticular JIA(pJIA)in patients aged ﹥ 2 years old. The approval was based on the favorable results from 2 randomized,double - blind,placebo - controlled,multinational,phase Ⅲ trials conducted in patients aged 2 - 17 years old with active sJIA or pJIA. Tocilizumab was generally well tolerated in patients with sJIA and pJIA. The most frequently reported adverse events in Tocilizumab recipients were infections,neutropenia,impaired liver function,etc.

OBJECTIVETo detect plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) in acute Kawasaki disease (KD) and analyze the relationship between NT-proBNP and other bio-markers in order to evaluate if NT-proBNP could be as a useful diagnostic marker to predict the risk of coronary arterial lesions in acute KD.

METHODTotally 106 patients with KD were recruited from January 2012 to April 2014 at Department of Pediatrics of Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology,64 were boys and 42 were girls, their age ranged from 2 months to 8 years and 4 months. Of the 106 cases, 48 had typical KD(TKD) and 58 incomplete KD(IKD). They were divided into two groups according to echocardiography results: coronary arterial lesions (KD-CAL, n = 33) and non coronary arterial lesions (KD-nCAL, n = 73). Forty children whose age and gender matched with respiratory tract infection were selected as control group, 22 were boys and 18 were girls, age range from 7 months to 7 years and 11 months. Plasma NT-proBNP levels were measured by using the enzyme-linked fluorescence analysis (ELFA) at the day of admission, meanwhile blood routine tests, liver function tests, determination of C-reactive protein (CEP), erythrocyte sedimentation rate (ESR), electrolytes were performed in these patients. Pearson's correlation analysis was used to evaluate the association. The ROC curve analysis was done to identify the threshold of coronary 'arterial lesions.

RESULTThe levels of NT-proBNP were (1 037 271) ng/L in TKD group and (1,325 ± 264) ng/L in IKD group. The levels of NT-proBNP in control group was (125 ± 22) ng/L. Both the levels of NT-proBNP in TKD and IKD group were significantly higher than that of control group (t = 3.360, 3.590; P < 0.05). The level of NT-proBNP in KD-CAL group was (2,775 ± 842) ng/L and that of KD-nCAL group was (830 ± 145) ng/L, NT-proBNP levels of KD-nCAL group was significantly higher than that of control group (t = 3.660, P = 0.007) ; moreover the level of NT-proBNP of KD-CAL group was also significantly higher than that of KD-nCAL group ( t = 3.860, P = 0.005). The levels of total white blood cell count, neutrophil percentage, platelet count, CRP and ESR of KD-CAL group were significantly higher than those of the control group, however there was no significant difference between KD-CAL group and KD-nCAL group. The levels of albumin and Na of KD-nCAL group were significantly lower than those of the control group. Plasma NT-proBNP level in KD group was positively correlated with white blood cell count, neutrophil percentage, and CRP (r = 0.239, P = 0.025; r = 0.359, P = 0.001; r = 0.474, P = 0.001), there was a negative correlation between albumin and Na (r = -0.303, P = 0.015; r = -0.338, P = 0.002). When the level of NT-proBNP was higher than 950 ng/L, the sensitivity for diagnosis of coronary arterial lesions in the KD was 88.1% and the specificity was 89.0%.

CONCLUSIONThe plasma NT-proBNP can be used as a useful parameter in early diagnosis of KD. Plasma NT-proBNP could be used to predict the risk of coronary arterial lesions in KD.

Biomarkers ; Blood Sedimentation ; C-Reactive Protein ; Case-Control Studies ; Child ; Child, Preschool ; Coronary Vessels ; pathology ; Female ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Predictive Value of Tests ; ROC Curve ; Sensitivity and Specificity

To investigate the effect of preceding naloxone injection into the third cerebroventricle or acute subdiaphragmatic vagotomy on the gastric acid secretion inhibited by the somatostatin analogue octreotide given by intracerebroventricular (icv) injection. The third ventricles were cannulated in male Wistar rats anesthetized with sodium pentobarbital. One week later, acute gastric lumen perfusion was carried out. The gastric perfusion samples were collected every 10 min and were titrated by 0.01 mol/L NaOH to neuter. On the basis of subcutaneous injection of pentagastrin (G-5, 160 micro, g/kg), icv injection of physiological saline (group A, n = 20), icv injection of octreotide (0.05 micro g) (group B, n = 20), icv injection of naloxone (2.5 micro g)+octreotide (0.05 micro g) (group C, n = 20), acute subdiaphragmatic vagotomy+ icv injection of physiological saline (group D, n = 20), or acute subdiaphragmatic vagotomy+icv injection of octreotide (0.05 micro g) (group E, n = 20) were conducted. Before and after icv injection, 1-h total acid output (TAO) was determined and compared. The experimental data were expressed in change rate (%) of TAO. The change rates (%) of TAO were 4.60% in group A, -20.35% in group B, -18.06% in group C, 5.01% in group D and -21.59% in group E, respectively. Comparison of group B or C versus group A showed that P < 0.01 and comparison between the group E versus group D showed that P < 0.01. Whereas the differences between group C and group B, group E and group B were not statistically significant (P > 0.05 for all). The results indicate that the central inhibition of gastric acid secretion by octreotide may not be mediated by the endogenous opiate substance or its receptor and the peripheral pathway for icv injection of octreotide to suppress gastric acid secretion is via extra-vagus route.

Objective To evaluate the effect of airway pressure release ventilation (APRV) in patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS), to evaluate the extent of ventilator-induced lung injury (VILI), and to explore its possible mechanism. Methods A prospective study was conducted in the Department of Critical Care Medicine of the First Hospital of Hebei Medical University from December 2010 to February 2012. The patients with ALI/ARDS were enrolled. They were randomly divided into two groups. The patients in APRV group were given APRV pattern, while those in control group were given lung protection ventilation, synchronized intermittent mandatory ventilation with positive end-expiratory pressure (SIMV+PEEP). All patients were treated with AVEA ventilator. The parameters such as airway peak pressure (Ppeak), mean airway pressure (Pmean), pulse oxygen saturation (SpO2), mean arterial pressure (MAP), heart rate (HR), central venous pressure (CVP), arterial blood gas, urine output (UO), the usage of sedation and muscle relaxation drugs were recorded. AVEA ventilator turning point (Pflex) operation was used to describe the quasi-static pressure volume curve (P-V curve). High and low inflection point (UIP, LIP) and triangular Pflex volume (Vdelta) were automatically measured and calculated. The ventilation parameters were set, and the 24-hour P-V curve was recorded again in order to be compared with subsequent results. Venous blood was collected before treatment, 24 hours and 48 hours after ventilation to measure lung surfactant protein D (SP-D) and large molecular mucus in saliva (KL-6) by enzyme linked immunosorbent assay (ELISA), and the correlation between the above two parameters and prognosis on 28 days was analyzed by multinomial logistic regression. Results Twenty-six patients with ALI/ARDS were enrolled, and 22 of them completed the test with 10 in APRV group and 12 in control group. The basic parameters and P-V curves between two groups were similar before the test. After 24 hours and 48 hours, mechanical ventilation was given in both groups. The patients' oxygenation was improved significantly, though there were no significant changes in hemodynamic parameters. The Pmean (cmH2O, 1 cmH2O = 0.098 kPa) in APRV group was significantly higher than that in control group (24 hours: 24.20±4.59 vs. 17.50±3.48, P < 0.01; 48 hours: 18.10±4.30 vs. 15.00±2.59, P < 0.05). After ventilation for 24 hours, the ratio of patients with increased Vdelta in APRV group was higher than that in control group (90% vs. 75%), but without statistical difference (P > 0.05). The SP-D level (μg/L) in serum in APRV group showed a tendency of increase (increased from 19.70±7.34 to 27.61±10.21, P < 0.05), in contrast there was a tendency of decrease in control group (decreased from 21.83±7.31 to 16.58±2.90, P > 0.05), the difference between the two groups was statistically significant (P < 0.05). After 48-hour ventilation, SP-D in APRV group was decreased, but no change was found in control group, and no significant difference was found as compared with that of the control group (16.45±8.17 vs. 17.20±4.59, P > 0.05). There was no significant difference in serum KL-6 between the two groups before and after ventilation. The SP-D and KL-6 levels in serum were unrelated with 28-day survival rate of the patients. The odds ratio (OR) of SP-D were 0.900 [95% confidence interval (95%CI) = 0.719-1.125], 1.054 (95%CI = 0.878-1.266), 1.143 (95%CI = 0.957-1.365), and the OR of KL-6 were 1.356 (95%CI = 0.668-2.754), 0.658 (95%CI = 0.161-2.685), 0.915 (95%CI = 0.350-2.394) before the test, 24 hours and 48 hours after ventilation (all P > 0.05). Conclusions APRV was similar to lung protective ventilation strategy in oxygenation and improvements in the lung mechanics parameters. APRV with a higher Pmean can recruit alveolar more effectively, and it had no impact on hemo-dynamics, but might exacerbate VILI.

Objective To investigate the effect of different sex hormones on insulin-stimulated glucose uptake on cultrured 3T3-L1 preadipocytes and adipocytes.Methods We treated cultured 3T3-L1 preadipocytes and adipocytes with different concentrations of 17?-Estradiol or testosterone and the effects were assessed as 2-deoxy glucose uptake.Results After incubation with 17?-Estradiol or testosterone,the ability of insulin-stimulated glucose transport was statistically significantly reduced.Dosage response study demonstrated that(10~(-7) mol/L) testosterone or(10~(-8) mol/L) 17?-Estradiol can induce insulin resistance(P

Objective To construct an eukaryotic expression vector of the hepatitis C virus(HCV)NS5A gene.and obtain a stable transfected Huh-7 cell line which provides a basis for further investigation of the hepatitis virus C NS5A protein.Methods The HCV NS5A gene from the pcDNA3.1(+)/HCV NS345 plasmid with HCV NS5A gene was amplified by PCR,and cloned to pGEM-T vector,and transformed into E.coli JM109.The positive colonies were first confirmed by restriction enzyme digestion and sequencing and then were inserted to eukaryotic expression vector pCI-neo,and verified by enzyme digestion and sequencing.Results After TA colon of the HCV NS5A was amplified by PCR,the positive colonies were finally verified by sequencing,which were totally in line with the designed coding sequence of HCV NS5A gene.Conclusion Eukaryotic expression vector of HCV NS5A gene has been successfully constructed.

Objective To explore the technique and feasibility of using 1.5 T MR guided radiofrequency ablation (RFA) of hepatic malignant tumor. Methods Twenty three patients with 44 malignant lesions in liver confirmed by pathology were treated with 1.5 T MR guided RFA using MR compatible multipolar RF electrode. Only patients refusing open surgery or suffering from unresectable lesions were included. Of these, 11 patients had primary hepatic carcinoma and 12 patients had hepatic metastases. The mean maximal diameter of lesions was (3.3 ± 1.8)cm. Postoperative MR was performed; the ablation zone covered and exceeded 0.5 to 1.0 cm to the margin of initial tumor was considered successful. Results All ablations were successful and lesions created by radio frequency were large enough to cover the initial tumor volume in all cases. No severe complications such as biliary fistula, perforation of diaphragmatic muscle,postoperative jaundice and pneumothorax were encountered. The mean operative time was (93 ± 33 ) min.The RF electrodes appeared in MRI as low signal structure. The ablation lesions were well-defined hyperintensity in T1 WI and hypo-intensity with a thin rim of high signal intensity on T2WI and DWI. Conclusion 1.5 T MR guided RFA of hepatic malignant tumor is an effective and safe technique.

AIM: To evaluate the potential acylation stimulating protein (ASP) resistance in both adipocytes and preadipocytes under the conditions by which insulin resistance is produced by the stimulation of free fatty acids (FFA), and to explore the mechanism of ASP resistance on post-receptor level. METHODS: 3T3-L1 preadipocytes were induced to differentiate. Then the cells were treated with oleate or palmitate at concentration of 0 mmol/L (FFA-free DMEM/F12), 0.125 mmol/L, 0.5 mmol/L or 1.0 mmol/L overnight. Glucose transport was assessed by [~3H] 2-deoxyglucose uptake to evaluate insulin resistance and ASP resistance. Both non-FFA treated and FFA treated 3T3-L1 cells were cultured with ASP at concentration of 5.0 μmol/L for 4 h, then the cell proteins were extracted, and the expressions of guanine nucleotide binding protein beta (Gβ), guanine nucleotide-binding protein alpha-q/11(Gαq/11), phosphorylated-protein kinase Cα (p-PKCα) and phosphorylated-protein kinase Cζ (p-PKCζ) were measured by Western blotting. RESULTS: Both adipocytes and preadipocytes were responsive to ASP. ASP stimulation increased glucose transport by 198% in adipocytes and by 287% in preadipocytes (P<0.01 vs PBS). FFA at concentration of 0.125 mmol/L did not change ASP-stimulated glucose transport significantly, but high dose of oleate or palmitate effectively reduced the ASP response with a significant reduction by 47% (P<0.05 for oleate) and 34% (P<0.05 for palmitate) at 1 mmol/L FFA in adipocytes. Similarly in preadipocytes, glucose uptake rates were decreased by 43% (P<0.05 for oleate) and 62% (P<0.01 for palmitate) at 1 mmol/L FFA. Effects were comparable to those obtained with insulin. After overnight incubation with oleate or palmitate in adipocytes and preadipocytes, Gβ, Gαq/11, p-PKCα and p-PKCζ were downregulated both in the absence of ASP treatment and in the presence of ASP treatment in adipocytes. At concentration of 1.0 mmol/L, oleate inhibited the expressions of ASP-induced Gβ, Gαq/11, p-PKCα and p-PKCζ in adipocytes by 47%, 44%, 39% (P<0.05, P<0.01) and 20% (P>0.05), respectively. Palmitate also effectively blocked the expressions of ASP (at concentration of 1.0 mmol/L)-induced Gβ, Gαq/11, p-PKCα and p-PKCζ by 50%, 43%, 44% and 43% (P<0.05, P<0.01) in adipocytes. In preadipocytes, oleate only inhibited ASP-induced p-PKCα and p-PKCζ significantly by 39% and 19%, respectively (P<0.05). However, overnight exposure of 3T3-L1 preadipocytes to 1 mmol/L palmitate leaded to 45%, 50%, 52% and 21% (P<0.05, P<0.01) inhibition of ASP-induced expressions of Gβ, Gαq/11, p-PKCα and p-PKCζ, respectively. CONCLUSION: Oleate and palmitate inhibit ASP-mediated stimulation of glucose transport both in adipocytes and preadipocytes. The study provides direct evidence of ASP resistance under the condition of insulin resistance induced by FFA in a cellular model. The mechanism of action involves both changes in expression of C5L2 as well as signaling parameters. Fatty acid-induced ASP resistance may contribute to the physiological abnormalities associated with insulin resistance and obesity phenotype.

In order to investigate the clinicopathological characteristics of aortic valve disease in children, all the native surgically excised aortic valves obtained between January 2003 and December 2005 were studied macroscopically and microscopically. The patients' medical records were reviewed and the clinical information was extracted. According to preoperative echocardiography, intraoperative assessment, and postoperative pathology, combined with clinical symptoms and signs, aortic valve diseases were divided into three categories: aortic stenosis (AS), aortic insufficiency (AI), and aortic stenosis with insufficiency (AS-AI). The etiology was determined according to the macroscopic, microscopic and clinical findings. The results showed that among 70 aortic valves, patient age ranged from 6 to 18 years, with a mean of 15.4 years, and there were 56 boys and 14 girls (male: female=4:1). Forty-four children only had pure aortic valve disease, and the other 26 children had aortic valve disease associated with other heart valve diseases. There were 5 cases of AS (7.14%), 60 cases of AI (85.71%) and 5 cases of AS-AI (7.14%). The causes were congenital aortic valve malformation (32 cases, 45.71%), rheumatic disease (28 cases, 40%), infective endocarditis (7 cases, 10%), Marfan syndrome (2 cases, 2.86%), and undetermined (1 case, 1.43%). It was concluded that the common causes of aortic valve disease in order of frequency in children were congenital aortic valve malformation, rheumatic disease, infective endocarditis, and Marfan syndrome. AI was more common in children with aortic valve disease. Compared with adult patients, congenital bicuspid aortic valve in children was often AI. Histologically, the leaflets of congenital bicuspid aortic valve were mainly myxomatous, fibrosis and calcification less seen. AI was frequently found in rheumatic disease, mostly associated with other heart valve diseases. Macroscopic and microscopic examinations together with clinical information, echocardiographic findings and operative details were important in evaluating the etiology of aortic valve disease.

AIM: The purpose of the present study was to examine the effect of insulin and different kinds of free fatty acid (FFA) on glucose transport in cultured 3T3-L1 preadipocytes or adipocytes. METHODS: Following the exposure of preadipocytes to insulin at different concentrations and treated times, glucose transport was assessed as [3H]2-deoxy glucose uptake. Furthermore, 3T3-L1 preadipocytes and adipocytes with either the monounsaturated FFA oleate (C18:1) or the saturated FFA palmitate (C16:0)were used and glucose transport was examined as above. RESULTS: Insulin increased specific membrane glucose transport in 3T3-L1 preadipocytes at the time of 15 min to 1 h stimulation. However, after 6 h exposure to insulin, downregulation of glucose transport was observed. Dose response studies demonstrated that 2-DG transport increased by 336% at 50 nmol/L of insulin (P