AIM:To study functions of pro-domain in tumor necrosis factor-? converting enzyme (TACE) maturation and to develop an approach to interfere with inflammation processes. METHODS: The plasmid pIRES2-EGFP was used to generate the expression plasmids constructed with the signal peptide and pro-domain, full length or without the pro-domain(designated as pIRES2-EGFP/T648, pIRES2-EGFP/T2472, pIRES2-EGFP/T57-T1824,respectively). The plasmids were used to transfect the U937 cells.After the transfected cells were stimulated by LPS, the effect of expression plasmids on TNF-? secretion was detected by ELISA and flow cytometry(FCM).RESULTS: The plasmid pIRES2-EGFP/T648 inhibited 61.09% TNF-? secretion and mediated the accumulation of TNF-? on the surface of U937 cells.The sTNF-? secretion and the level of the mTNF-? in the plasmid pIRES2-EGFP/T57-T1824 transfected cells had no significant deviation compared with the pIRES2-EGFP transfected cells. The sTNF-? secretion of the cells transfected by the plasmid pIRES2-EGFP/T2472 increased and the level of the mTNF-? decreased.CONCLUSION: The pro-domain has dual effects in TACE maturation and might be applied to molecular design of anti-inflammation drug.

Objective To develop an albumin aptamer that may potentially serve as a selective ligand for albumin removal from experimental samples.Methods A single-stranded 59nt DNA library that contains 21 random oligo nucleotides was synthesized in vitro.An albumin aptamer A6 was developed by SELEX technique using bovine serum albumin (BSA) as target.The enrichment of aptamer and evaluation of its binding properties were monitored by flow cytometry.The secondary structure of A6 was predicted by MFord software.Results The aptamer A6 strongly bound to BSA with a Kd of 77.4 nmol/L,and had minimal cross reactivity with control proteins including ovalbu min,IgG,and trypsin.Conclusions Aptamer A6 may be a potential tool in albumin removal.

Objective To review the clinical characteristics and treatment of type 1 diabetes mellitus (T1DM) in children. Methods The clinical data of 103 children with T1DM admitted to our hospital from Februry 2002 to August 2010 were retrospectively analyzed. Thirty one cases with diabetic ketoacidosis (KDA) were treated with continuous insulin pump (group A) or basal-bolus insulin therapy (group B). The differences in blood glucose control time, the rate of hypoglycemic episodes, glucose fluctuation, fasting blood glucose (FBG), 2 h postprandial blood glucose (2 hPBG), insulin dosage, the time of urine acetone bodies disappear and length of stay were compared in two groups. Results The age of 103 children with T1DM was from 38 d to 15. 33 y with an average of (8 ±3) y; most of them was 7 - 10 y (47, 45.6% ). Seventy eight children were first diagnosed accounting for 75.7%; boys accounted for 55.3% of total. Fifty one cases (49.5%) were diagnosed in winter and spring and 67 (65.2%) had infections, most of them were respiratory tract and gastrointestinal infections. Sixty two cases (60. 2% )presented as diabetic ketoacidosis at the first onset, including 4 cases (3.9%) with cerebral edema. Some patients were complicated by Hashimoto's thyroiditis, hyperthyroidism, SLE and other autoimmune diseases.Among 31 cases with ketoacidosis the FBG and PBG were decreased significantly after treatment, there were no significant differences between two groups (P ＞ 0. 05 ). Compared to group B the correction time of DKA and urine acetone bodies was shorter, and reached the targeted glucose levels more quickly, the frequency of blood fluctuation and the hypoglycemia was significantly lower, the length of stay was shorter, and the daily dose of insulin was lower in group A; the differences between two groups were statistically significant ( P ＜0. 05 or P ＜0. 01 ). Conclusions The clinical symptoms at first onset of T1 DM in children are not typical,and likely to be combined with DKA; infection may be one of the inducing factors for DKA. Continuous subcutaneous insulin infusion with pump can control the blood glucose more effectively and equably, and are convenient for use by children; so it is a better treatment option for type 1 diabetes mellitus in children.

Objective To study the expressions of miRNAlet7 and Ras in non-small cell lung cancer ( NSCLC),and their correlations with clinicopathological features and survival time.Methods In-situ hybridization was used to detect the expression of let7,and SP immunohistochemistry to measure HMGA2 in 68 NSCLC cases ( group A) and 20 cases with normal lungs ( group B).Results The positive rate of let7 in group A was lower than that in group B (39.7％ vs 63.2％ ) ( P ＜0.05).The positive rate of Ras in group A was higher than that in group B (66.2％ vs 25.0％ ) ( P ＜0.01 ).The positive rate of let7 was not related to the age,gender,histological type,cell differentiation,and clinical stages of cancer patients(P ＞0.05).The positive rate of Ras was related to smoking,sex,and histological type of cancer( P ＜0.01 ),and was not related to cell differentiation,lymphatic metastasis,and clinical stages of cancer ( P ＞0.05).There was an obvious negative correlation between let7 and Ras( r =-0.627,P ＜0.01 ).The 2-year survival rate of let7-positive group was higher than that of the let 7-negative group ( x2 =4.84,P ＜0.05).No statistically significant difference was found between Ras-positive and-negative groups ( P ＞0.05 ).Conclusions The lower level of let7 expression,and high level of Ras expression have much to do with the carcinogenesis of NSCLC.The level of let7-positive expression is closely related to prognosis; while Ras may act in cooperativity in the occurrence and development of NSCLC.

With the rapid development of medical technology,burn treatment level is also rising.However,large area bum injury with inhalation injury treatment has not yet made a breakthrough,and become the main cause of death in burn patients.Severe shock,infection,multiple organs failure and acute respiratory system injury caused by inhalation injury are the main factors threatening the life of patients.The treatment of large area burn combined with inhalation injury has been the focus of bum and clinical research.Domestic and foreign scholars on large area bum associated with inhalation injury treatment for a large number of studies.In recent years,with the deepening of research,some new treatment concepts have been proposed.The advanced treatment progress of large area burn combined with inhalation injury were reviewed.

A novel method for simultaneous detection of mycotoxins (e.g.,aflatoxin B1) or their metabolic residues in animal plasma with impurity adsorption purification followed ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed.Extraction of mycotoxins and their metabolites from animal plasma sample was performed with 0.1％ formic acid-acetonitrile solution after addition of sodium chloride and hydrous magnesium sulfate.The extract was then dehydrated and purified with hydrous magnesium sulfate,C18,primary secondary amine,and alumina-A.3 mL of the supernatant was evaporated and re-dissolved with 0.5 mL of 0.1％ formic acid aqueous solution/acetonitrile (70∶ 30,V/V) for UPLC-MS/MS detection.The analytes were separated by a C18 column utilizing gradient elution with 0.1％ formic acid aqueous solution containing 0.5 mmol of ammonium acetate and 0.1％ formic acid-methanol solution,and finally detected by tandem mass spectrometry in positive/negative ESI mode.Identification and quantification were achieved by LC-MS/MS with multi-reaction monitoring (MRM).Good linearity in response was obtained in the analytes concentration range of 0.05-100 ng/mL with correlation coefficients larger than 0.99.The limits of quantification (S/N=10) were around 0.05-0.5 ng/mL.The recoveries of mycotoxins and their metabolites spiked in blank plasma samples were in the range of 62.0％-116.4％,with relative standard deviations (RSDs) less than 19.0％.

Objective To assess the local treatment efficacy of percutaneous radiofrequency ablation(PRFA) in combination with percutaneous ethanol injection(PEI) therapy protocol for liver metastases in patients with colorectal cancer.Methods From February 2000 to February 2004, total of 61 liver metastatic lesions(1.5～7.8 cm) from 37 colorectal cancer patients were ablated by using RF-2000 generator and 10-needle LeVeen electrode percutaneously guided by B-ultrasound.Tumors larger than 2.5 cm in diameter were treated by multiple overlapping radiofrequency ablations encompassing the entire lesion as well as a rim of normal liver tissue(approximately 1.0 cm).PEI was performed additionally.The Chiba needle and Quadra-Fuse multi-pronged injection needle were used for tumors ranged 1～2 cm and for those larger than 3 cm respectively.Results The procedures were performed successfully in all patients.The post-PRFA/PEI complication rate was 5.4%.The follow-up rate was 97.3%(36/37).Thirteen patients achieved a complete response(CR),and 21 achieved a partial response(PR).The effective rate(CR+PR) was 94.4%.The patients' survival rates were 97.2% at 1 year,75.0% at 2 years,and 52.8% at 3 years respectively.Conclusion The results of this study suggest that the treatment protocol of PRFA in combination with PEI was an effective mini-invasive local ablation therapy,which was part of the comprehensive management of the colorectal liver metastases.

ObjectiveTo investigate reproducibility of cardiac MRI for assessment of cardiac morphology and ventricular function in selected normal Chinese Han population.MethodsTwo hundred and sixty-nine normal volunteers underwent cardiac MRI using a 1.5 T MR system.HASTE and steady state free precession imaging were performed with long and short axis images and cine mode through the ventricle with wireless vector cardiac gating.The images were reviewed by two independent observers.The dimensions of cardiac chambers and ventricular function including ejection fraction (EF),end diastolic volume( EDV),end systolic volume (ESV) and myocardial mass were evaluated.The data between male and female were compared by using two-tailed unpaired t test.Results Total imaging time was (15 ± 3 ) min.The anteroposterior diameter of the left atrium was( 2.87 ± 0.77 ) cm,the right atrial diameter perpendicular to the atrial septum was ( 3.61 ± 0.57 ) cm,the end diastolic diameter of the left ventricle was (4.97 ±0.52) cm,the end diastolic diameter of the right ventricle was (2.65 ± 0.48) cm.On the left ventricle,EF was(60.62 ±7.08)％,EDV was (115.37 ±26.71) ml,ESV was (46.02+15.72) ml and LV mass was ( 82.97± 24.03 ) g.On the right ventricle,EF was (47.73 ± 6.50 ) ％,EDV was ( 128.27 ± 32.16 )ml,ESV was (67.7 ± 21.07) ml and RV mass was (48.24 ± 13.42) g.There were no statistically significant differences in LVESV( P =0.144 ),LVEDV index ( P =0.714 ),LVESV index ( P =0.113 ),LVCI ( P =0.199),RVEF ( P =0.296 ) and RV mass ( P =0.093 ),and statistically significant differences in other cardiac parameters between male and female.Conclusion Cardiac MRI can provide useful information about cardiac function and morphology with a high level of reproducibility in normal Chinese Han population.

OBJECTIVETo assess the association between single nucleotide polymorphisms (SNPs) of calcium channel β 2 subunit (CACNB2) gene and essential hypertension (EH) in ethnic Han Chinese in Wenzhou area, and to study the influence of rs7069292 alleles on gene expression with luciferase reporter technique.

METHODSSixty hundred and thirty seven Han Chinese with EH and 600 normal controls were enrolled. Genotypes of 6 SNP within CACNB2 gene including rs2228645, rs2357928, rs7069292, rs7099380, rs10764319 and rs11014166 were determined with matrix assisted laser desorption ionization/time of flight mass spectrometer (MALDI-TOF MS). A luciferase reporter gene plasmid containing the fragment flanking rs7069292 (-2831 bp to -2460 bp) in the 5' regulatory region of CACNB2 was constructed.

RESULTSCompared with the control, CT and TT genotypes for the rs7069292 locus were significantly more common in EH group (5.20% vs. 2.17%, 2.59% vs. 1.08%, P< 0.05). CC genotype was not found. Promoter activity for allele C of the rs7069292 locus was significantly increased compared with allele T (P< 0.05). No significant difference was detected for other 5 SNPs in terms of genotypes and allele frequency.

CONCLUSIONThe rs7069292 CT polymorphism of the CACNB2 gene is associated with EH in ethnic Han Chinese from Wenzhou area. A T>C mutation may affect the expression of CACNB2.

Aged ; Alleles ; Base Sequence ; Calcium Channels, L-Type ; genetics ; Case-Control Studies ; Cell Line ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Objective:To investigate prognostic factors for acral lentiginous melanoma (ALM) , and to construct a nomogram to verify the predictive value of these factors.Methods:Clinical data on 1 573 patients with ALM were collected from the Surveillance, Epidemiology, and End Results (SEER) database of National Cancer Institute in United States between 2004 and 2015. Data about patients′ age, gender, ulcer status, SEER staging, surgical protocols, T-, N- and M-staging, overall survival rates and disease-specific survival rates were extracted. Chi-square test was used to analyze the correlation of clinical characteristics with overall survival rates and melanoma-specific survival rates, and univariate and multivariate Cox proportional hazards regression models were used to investigate prognostic factors and establish predictive models.Results:Among the 1 537 patients with ALM, 714 were males, 823 were females, 818 were under 64 years of age, and 1 363 were Caucasian. Skin lesions occurred on the lower limbs and buttocks in 1 205 cases, and 974 cases had ulcers; according to the SEER staging, non-spread localized skin lesions were observed in 1 048 cases. There were significant differences in the mortality rate among patients of different ages at diagnosis, different gender, with different ulcer status, surgical status, and at different SEER stages, T-stages, N-stages and M-stages (all P < 0.001) . Univariate and multivariate Cox regression analysis showed that age ≥ 65 years, male, ulcers and distant lymph node metastasis in the SEER staging were associated with increased risk of death in the patients (all P < 0.05) , and the mortality rate was significantly higher in the patients with T2-, T3- or T4-stage ALM than in those with T1-stage ALM (all P < 0.05) , and higher in the patients with N1-, N2- and N3-stage ALM than in those with N0-stage ALM (all P < 0.05) . Conclusion:Age, gender, ulcer status, SEER stage, T-stage and N-stage are independent prognostic factors for overall survival rates and disease-specific survival rates of ALM.