Anti-angiogenesis mechanism plays a vital role in tumor targeting immunotherapy. Based on the amino acid sequence of an anti-VEGFR-2 scFv-Fc fusion antibody (AK404R-Fc), this article is aimed to generate an anti-VEGFR-2 human IgG1-like full length antibody (Mab-04). Firstly, the light chain (L-chain) and heavy chain (H-chain) were obtained by overlap PCR and then linked to eukaryotic expression vector pcDNA3.1, separately. The recombinant plasmids (pcDNA3.1-L-chain and pcDNA3.1-H-chain) were then co-transfected into CHO-k cells using liposome transient transfection. Subsequently, Mab-04 antibody was expressed and purified by Protein A affinity chromatography. Western blotting was applied to identify the expression of Mab-04 and its affinity was detected by ELISA assay. DNA sequencing revealed the successful construction of recombinant plasmids and Western blotting assay proved the successful expression of full-length antibody (1 microg x mL(-1)). Finally, ELISA assay illustrated that the binding of the antibody to its antigen was in a concentration-dependent manner (IC50: 50 nmol x L(-1)). These outcomes above indicated that Mab-04 was successfully expressed and assembled, which laid the foundation for further preparation and antineoplastic activity study.

BACKGROUND:As a by-product in sugar industry, sugar cane wax has been widely used in non-medical field. Some researches indicate that sugar cane wax plays a great role in reducing blood cholesterol; however, the therapeutical effect and clinical application should be studied further.OBJECTIVE: To separate the high-class eicosanoic acid and the high-class alkanols, which are suitable for medical application, and further to observe the effect of them on reducing blood cholesterol of model rets with hyperlipemia.DESIGN: Randomized control animal study.SETTING: Pharmacological Institute, Chongqing Kangerwei Pharmaceutical Co., Ltd.MATERIALS.: The experiment was carried out in the Pharmacological Institute of Chongqing K.E.W Pharmaceutical Go.,Ltd. From April 2005 to January 2006. A total of 65 adult female Wistar rats, aged at 3-6 months, weighing 180-220 g, of SPF grade, were provided by Experimental Animal Center of Chongqing Institute of Traditional Chinese Medicine. Raw sugar cane wax was provided by Beijing Jiade Hongsheng Biochemical Technology Co., Ltd. High-class alkanols C26,C28,C30, C32 and high-class eicosanoic acid C28, C30, C32, C34 were provided by Sigma Company (standard materials of gas phase chromatography), and other reagents were national analytical pure.METHODS: ① Sugar cane wax was extracted from raw sugar cane wax with ethanol and other organic solution and separated from the mixture of high-class eicosanoic acid and the mixture of high-class alkanols with saponification and calcification. Main components were analyzed with gas phase chromatography. The main components of high-class alkanols were C26, C28, C30 and C32 and the main components of high-class eicosanoic acid were C28, C30, C32 and C34, ② Based on references, rats were fed in 3 days and randomly divided into blank group (n =10) and experimental group (n =55).And then, all rats were cut off their tails to collect blood and the triacylglyoerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were measured with automatic biochemistry analyzer. Rats were fed with common granule feeds in blank group or with high-lipid feeds (containing 0.1 mass fraction of oiliness, 0.1 mass fraction of yolk powder, 0.01 mass fraction of cholesterol, 0.002 mass fraction of pig's gall salt, 0.788 mass fraction of common feeds) in experimental group. All rats ate and drank freely. Seven days later, blood was collected again from tail tip to measure the contents of TG, TC and HDL-C. Based on level of serum TG, rats in the experimental group were randomly divided into 5subgroups (n =11): negative control group, low-dosage high-class alkanols group, high-dosage high-class alkanols group,Iowdosage high-class eicosanoic acid and high-dosage high-class eicosanoic acid group. Rats in low-dosage and high-dosage high-class alkanols groups were perfused with 5 and 50 mg/(kg·d) high-class alkanols; meanwhile, rats in low-dosage and high-dosage high-class eicosanoic acid groups were perfused with 20 and 200 mg/ (kg·d) high-class eicosanoic acid. Rats in negative control group and blank group were perfused with the same volume of 0.3% carboxymethylcellulose sodium and distilled water, respectively, once a day for successive 30 days. At 16 hours after the last administration, rats were anesthetized to collect blood from heart to measure contents of TG, TC and HDL-C in serum.MAIN OUTCOME MEASURES: ① Percentage of main component in separated mixtures of high-class eicosanoic acid high-class alkanols; ② levels of serum cholesterol, HDL and TG.RESULTS: A total of 65 experimental rats were involved in the final analysis. ① Gas phase chromatography suggested that the content of C28 high-class alkanols was the most (73.6%), and other three kinds of high-class alkanols were counted for 5.3% (C26), 6.2% (C30) and 5.1% (C32), respectively. The total quantity was 90.2%. In the mixture of high-class eicosanoic acid, content of C28 high-class eicosanoic acid was the most (46.6%) and the other three kinds of high-class eicosanoic acid were counted for 16.7% (C30), 6.8% (C32) and 9.3% (C34), respectively. The total quantity was 79.3%. ②Levels of serum TC were (1.46±0.27), (1.66±0.33), (1.44±0.25) and (2.16±0.52) mmol/L in high-dosage and Iow-dosage high-class alkanols groups and high-dosage and Iow-dosage high-class eicosanoic acid groups, respectively, which were lower than those in negative control group [(2.52±0.83) mmol/L, P＜0.01]. Levels of HDL-C were (0.73±0.09), (0.71±0.07), (0.79±0.10) and (0.70±0.08) mmol/L in the four treatment groups, respectively, which were higher than those in negative control group [(0.58±0.13) mmol/L, P＜0.05-0.01].CONCLUSION: The high-class alkanols and the high-class eicosanoic acids separated from sugar cane wax made in China significantly have the activity of reducing blood cholesterol; however, the effect on decreasing TG is not obvious.

Objective:To analyze the dynamic variety of serum Nesfatin-1in the patients with status epilepticus (SE) , and to explore the clinical values of the serum Nesfatin-1in evaluation on the illness condition and short-term prognosis in the patients with epileptic seizure.Methods:A total of 43patients diagnosed as primary epilepsy were collected as the subjects and received the regular clinical therapy.The serum levels of Nesfatin-1of the patients were detected before treatment and at the end of the 1st week after treatment, the end of the 2nd week therapy after treatment and the end of the 1st month after treatment, respectively.Liverpool Seizure Severity Scale 2.0 (LSSS2.0) was used to evaluate in the patients.The correlations of the serum levels of Nesfatin-1of the patients at each time point with the scores of LSSS were analyzed.After one year of follow-up, all patients were divided into two groups according to the prognosis, which were survival group (31cases) and death group (12cases) .The risk factors of one-year prognosis were analyzed by multivariate Logistic regression analysis.The sensitivity and specificity of the serum values of Nesfatin-1to the one-year prognosis were analyzed by receiver operating characteristic (ROC) curve.Results:Compared with before treatment, the LSSS scores and the serum Nesfatin-1levels of the patients in two groups were decreased with the increasing of treatment time (P＜0.05) .The LSSS scores and the serum levels of Nesfatin-1of the patients were not different between two groups before treatment (P＞0.05) .The LSSS scores and the serum levels of Nesfatin-1of the SE patients in survival group were lower than these in death group from the end of the 1st week after treatment to the end of the 1st month after treatment (P＜0.05or P＜0.01) .The positive correlations of the serum levels of Nesfatin-1and the LSSS scores of the patients at each time point were confirmed (r=0.617-0.726, P＜0.05) .The serum high level of Nesfatin-1was the risk factor of the one-year prognosis analyzed by multivariate Logistic regression analysis.The ROC curve identified the cutoff level of serum Nesfatin-1 (2.7μg·L-1) to the one-year prognosis with the sensitivity of88.5％and specificity of 79.6％.Conclusion:The dynamic variety of serum Nesfatin-1can reflect the condition of illness, and the serum level of Nesfatin-1is decreased gradually with the prolongation of time, and has application value in predicting the prognosis of the patients with epileptic seizure.

The elucidation of vapor-liquid equilibrium (VLE) of the halogenated silane was necessary for the production of silicon derivatives, especially for methylvinyldichlorosilane, due to the lack of the relevant reports. Isobaric VLE for the system methyldichlorosilane-dimethyldichlorosilane-benzene and isobaric VLE of the three binary systems were measured with a new pump-ebulliometer at the pressure of 101.325 kPa. These binary compositions of the equilibrium vapor were calculated according to the Q function of molar excess Gibbs energy by the indirect method and the resulted VLE data agreed well with the thermodynamic consistency. Moreover, the experimental data were correlated with the Wilson, NRTL, Margules and van Laar equations by means of the least-squares fit, the acquired optimal interaction parameters were fitted to experimental vapor-liquid equilibrium data for binary systems. The binary parameters of Wilson equation were also used to calculate the bubble point temperature and the vapor phase composition for the ternary mixtures without any additional adjustment. The predicted vapor-liquid equilibrium for the ternary system was in a good agreement with the experimental results. The VLE of binary and multilateral systems provided essential theory for the production of the halogenated silane.

OBJECTIVETo detect genetic causes of Duchenne muscular dystrophy (DMD).

METHODSNext-generation sequencing was used to detect 6 DMD patients in whom no exonic deletions were detected by multiplex PCR. Sanger sequencing and multiplex ligation-dependent probe amplification were used to confirm the results.

RESULTSOne case was found to have deletions of exons 10 and 11, 1 had exons 16 and 17 duplication, 4 cases have 8 point mutations including c.2776C>T, c.5475delA, c.6391_6392delCA, IVS64+1G>A, c.2645A>G, c.5244G>A, c.7728T>C, c.8729A>T, c.8734A>G and c.8810G>A. The former 4 mutations are suspicious pathogenicity, the other 6 mutations are polymorphisms in population. Three novel mutations (IVS64+1G>A, c.6391_6392delCA (p.Q2131NfsX3) and p.Q926X (CAG>TAG) were not reported before.

CONCLUSIONNext-generation sequencing technology is a useful tool for the detection of deletion, duplication and point mutation, which is valuable for clinical application.

Adolescent ; Base Sequence ; Child ; Genetic Variation ; Humans ; Infant ; Molecular Sequence Data ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Sequence Analysis, DNA ; methods

OBJECTIVETo study the mental health status and its affecting factors in nurses of general hospitals.

METHODSFour hundred and seventy nurses in general hospitals were investigated with SCL-90, Self-Rating Life Events and coping style questionnaires.

RESULTSThe scores of SCL-90 factors, such as anxiety, obsessive-compulsion, phobia, somatization, were higher than the norm (P < 0.05). 22% of the nurses had two factors higher than 2, and 5.3% ever had suicidal idea. The total score of SCL-90 and some factorial scores in < 40 years age group were higher than in older age groups (P < 0.01), and in III grade hospital nurses were also higher than in II grade hospital nurses (P < 0.01). The scores of negative life events, negative coping style and work load from abnormal psychology group were higher than normal group, while positive coping score was lower than normal group (P < 0.05, P < 0.01). Stepwise regression analysis showed that, negative life events, coping style, hospital grade had predictive value for total score of SCL-90 and anxiety, depression etc factorial scores (P < 0.01). Age had negative predictive value for anxiety, hostility and phobia (P < 0.05). The total score from work events had predictive value for obsession and hostility, and positive events from work had predictive value for anxiety and the total score of SCL-90 (P < 0.01). The total interpersonal relationship events and negative interpersonal relationship events had predictive value for hostility and interpersonal sensitivity.

CONCLUSIONGeneral hospital nurses especially in ages < 40 group had higher incidences of mental health problems. The main influencing factors are negative life events, coping style, work load, interpersonal relationship, age and hospital grade.

Adult ; Female ; Hospitals, General ; Humans ; Mental Health ; Middle Aged ; Nurses ; psychology ; Social Support ; Surveys and Questionnaires ; Workload ; Young Adult

OBJECTIVETo investigate the effects of rabbit limbs ischemia/reperfusion on myocardial necrosis and apoptosis in vivo.

METHODSThirty-six healthy new zealand rabbits were randomly divided into 3 groups: (1) Sham group; (2) I/R(Ischemia/reperfusion) group; (3) RPostC (remote postconditioning) group. The activity of blood serum creatine kinase (CK) and lactate dehydrogenase (LDH) were measured at baseline, the end of ischemia after 60 min and 120 min of reperfusion respectively. The extent of myocardial ischemia and the scope of myocardial infarction were assessed by evans blue and Triphenyl tetrazolium chloride (TTC). The myocardial cell's apoptosis at the area of myocardial ischemia was estimated by Tunel. Protein expression of caspase-3, Bcl-2 and Bax in myocardial ischemic area were analyzed with the method of immunohistochemistry.

RESULTSCompared with I/R group, the myocardial infarct size and the CK activity were significantly reduced in RPostC group. The Tunel positive index of RPostC group in ischemic myocardium was significantly lower than that in I/R group (21.79% +/- 1.07% vs 35.81% +/- 1.10%, P < 0.05). Caspase-3 positive cells index was calculated with randomly selected five regions in each slide and then the positive cells in per hundred cells were calculated. The RPostC group of caspase-3 positive cells was significantly lower than that in I/ R group(25.03% +/- 1.16% as 39% +/- 2.43%, P < 0.05). Compared with the sham group, the Bax protein expression index and the Bcl-2 protein expression index of I/R group and RPostC group were increased. The Bax/Bcl-2 ratio of RPostC group decreased, while it was increased in I/R. Compared with the I/R group, the Bax protein expression and Bax/Bcl-2 ratio of RPostC group significantly reduced, but the expression index of Bcl-2 ratio was significantly increased, the differences were statistically significant.

CONCLUSIONLimbs ischemia/postconditioning could significantly reduce necrosis and apoptosis of ischemia/reperfusion myocardium. The mechanism of reducing the myocardial cell apoptosis may have relation to inhibiting the activation of pro-apoptotic gene caspase-3 and increased expression of Bcl-2.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Creatine Kinase ; blood ; Ischemic Postconditioning ; L-Lactate Dehydrogenase ; blood ; Lower Extremity ; Male ; Muscle, Skeletal ; blood supply ; Myocardial Reperfusion Injury ; pathology ; Necrosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; bcl-2-Associated X Protein ; metabolism

Objective To evaluate the interactive effects of fine particulate matter and temperature on non-accidental mortality of residents in Pudong, Shanghai. Methods Daily mortality, air pollutants and meteorological data from Jan 1^st.2016 to Dec 31^st.2017 were collected.Generalized additive Poisson regression models was used to estimate the effects of PM_2.5 pollution on daily mortality, bivariate response surface models and temperature stratified models were applied to examine the interaction of temperature with PM_2.5 on mortality. Results A total of 43 345 non-accidental deaths were included, daily mean PM_2.5 concentration was 39.1 μg/m³, daily mean temperature was 17.7 ℃.A 10 μg/m³ increase in the daily PM_2.5 at lag 1 day was associated with a 0.56%(95%CI:0.11%-1.01%), 0.49%(95%CI:-0.19%-1.18%) and 0.22%(95%CI:-1.14%-1.60%) increase in non-accidental, cardiovascular and respiratory mortality, respectively.Higher risks were identified for males and the older (≥65 years).The effect estimates per 10 μg/m³ increase in PM_2.5 in medium temperature level were 0.59%(95%CI:0.04%-1.14%)for non-accidental, 0.51%(95%CI:-0.32%-1.35%)for cardiovascular and 0.51%(95%CI:-0.32%-1.35%) for respiratory mortalities.The PM_2.5 effects were approximately 2-4 times higher in higher temperature level for non-accidental and cardiovascular mortalities compared with other temperature levels; for respiratory mortality, the PM_2.5 effects was approximately 2-fold higher in lower temperature levels than the medium, although the interactions between temperature and PM_2.5 were statistically insignificant. Conclusions Temperature may modify the effects of PM_2.5 on mortality in Pudong, Shanghai and the interactive pattern may be different across disease-specific mortalities.

BACKGROUNDThe prevalence of overweight and obesity in Chinese children and adolescents was increasing during the past few decades. The goal of this study was to investigate the effects of after-school exercise with or without diet restriction on total and central obesity, fitness level, and metabolic profile in overweight Chinese adolescents.

METHODSA ten-week weight loss trial was performed using a 2 × 2 block design (exercise × diet). Ninety-three overweight adolescents (average age: (13.6 ± 0.7) years; body mass index (BMI): 22.4 - 34.1 kg/m(2)) were randomly assigned to four groups: 1) diet (D); 2) exercise (EX); 3) diet plus exercise (DEX); and 4) overweight control (C). Caloric intake recipes were enacted based on individual age and corresponding ideal body weight. One-hour after-school exercise was performed once per day, four days per week for ten weeks. Changes of anthropometry, body composition, aerobic fitness, and metabolic biomarkers were determined.

RESULTSGroups D, EX and DEX had a significant decrease in BMI (P < 0.01) after the intervention. The percentage of body and truncal fat, and waist circumference were independently reduced by exercise (P < 0.05 and P < 0.01), but not diet. The decrease in body fat percentage was positively related with the exercise compliance (r = 0.34, P = 0.01). Exercise decreased truncal fat percentage and waist circumference, suggesting a reduction of central adiposity, but did not significantly affect body weight and BMI. Exercise significantly reduced serum low-density lipoprotein cholesterol (P = 0.037), which was positively correlated with decreases of truncal fat percentage (r = 0.222, P = 0.048). No significant effects of interventions on insulin sensitivity, early insulin release index, and aerobic fitness were observed.

CONCLUSIONAt least twice a week of one-hour after-school exercise significantly attenuated central adiposity and had a significant impact on lipid profiles in overweight Chinese adolescents.

Adiposity ; physiology ; Adolescent ; Asian Continental Ancestry Group ; Body Composition ; physiology ; Body Mass Index ; Body Weight ; physiology ; Child ; Exercise ; physiology ; Female ; Humans ; Male ; Overweight ; diet therapy ; metabolism ; therapy ; Waist Circumference

OBJECTIVETo investigate the risk factors for selective devascularization in patients with portal hypertension.

METHODSThe clinical data of 160 patients with portal hypertension underwent selective devascularization were retrospectively analyzed. All the patients were divided into high-risk group and low-risk group according to the postoperative complications. Thirty-two clinical factors were analyzed using logistic regression.

RESULTSSingle-factor analysis showed that history of jaundice, Child-Turcotte-Pugh classification, total bilirubin (before the operation), prolongation of prothrombin time, pre-operative free portal pressure, ascites, leukocyte count (1 week after the operation) and hemoglobin (1 week after the operation) were significantly different between the high-risk group and low-risk group (P < 0.05). Logistic regression analysis showed that decrease of free portal pressure, total bilirubin (before the operation), prolongation of prothrombin time, ascites, leukocyte count (1 week after the operation) and hemoglobin (1 week after the operation) were still significantly different between the two groups (chi2 = 53.337, P < 0.01).

CONCLUSIONSThe risk factors of selective devascularization in patients with portal hypertension are decrease of free portal pressure, pre-operative total bilirubin, prolongation of prothrombin time, ascites, post-operative leukocyte count and hemoglobin.

Adult ; Aged ; Female ; Humans ; Hypertension, Portal ; surgery ; Logistic Models ; Male ; Middle Aged ; Portasystemic Shunt, Surgical ; adverse effects ; methods ; Postoperative Complications ; etiology ; Prognosis ; Retrospective Studies ; Risk Factors ; Young Adult