Objective To evaluate the relationship of the levels of serum oxide and antioxide with endothelium-dependent vasodilatation(EDV) in obese subjects and to explore the effect of oxidative stress on endothelial dysfunction in obese subjects. Methods 20 euglycemic obese males (Ob) and 13 age- matched normal controls (NC) underwent euglycemic hyperinsulinemia clamp study to evaluate the peripheral glucose disposal rate (GDR) in steady-state and brachial artery ultrasound studies to assess the endothelium-dependent vasodilatation (EDV). The serum levels of ROS, MDA, GSH-PX, GSH and free fatty acids (FFAs) were measured. Results The serum ROS, MDA and FFA concentrations were significantly higher in Ob group than in the controls (P

OBJECTIVETo construct and express the recombinant human adiponectin (gAd) global domain.

METHODSgAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG.

RESULTSThe gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis.

CONCLUSIONThe recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.

Adiponectin ; biosynthesis ; genetics ; Adult ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Female ; Genetic Vectors ; genetics ; Humans ; Prokaryotic Cells ; cytology ; metabolism ; Recombinant Proteins ; biosynthesis

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

Objective To evaluate the surgical treatment for renal cell carcinoma with inferior vena cava tumor thrombus and the clinical significance of multidisciplinary treatment. Methods Two cases of renal cell carcinoma with inferior vena cava thrombus diagnosed by Doppler ultrasonography and CT were included in this retrospective analysis. The tumor thrombus was in level Ⅱ in one case and in level Ⅳ in the other. Coagulation test and complete blood count were done again before surgery. Human albumin, fibrinogen, prothrombin complex, plasma, platelet, UW and irrigating solution were prepared before the operation.Under general anesthesia, surgery was performed using abdomen inverted Y shaped incision. Right radical nephrectomy was finished by the urological surgeon; the vena cava was completely dissected from the renal vein level to the secondary porta of the liver by the hepatobiliary surgeon, the vena cava and the surrounding branch vein were blocked in the upper and lower vena cava tumor thrombus; tumor thrombus was removed completely by the vascular surgeon. In one case (patient with level Ⅳ thrombus ) where the tumour thrombus invaded the wall of the vena cava, the thrombus was found to be extending to the cavo-atrial junction but not into the right atrium. The left femoral venous-right atrial bypass was established, the cardiopulmonary bypass lasted for 241 mia, and the aorta was blocked for 18 min. Salvage autotransfusion was used during surgery, and the hepatic vein of the secondary liver porta was anastomosed to artificial vascular graft.The data for surgical indication, operation time, operative blood loss and postoperative hospital stay were analyzed. Results Right radical nephrectomy and inferior vena cava thrombectomy were performed successfully, and the two patients were discharged on the 15th and 27th day after surgery, respectively. The two patients were followed up for 1 and 16 months after surgery, respectively, and both survived without local recurrence and distant metastasis. Conclusion Radical nephrectomy and inferior vena cava thrombectomy is the preferred method for patients without metastasis, and multidisciplinary cooperation could shorten the operation time, reduce the tumor recurrence and increase the survival rate of patients.

OBJECTIVETo understand the tendency of smoking in middle school students during the last 5 years and to identify the relationship between subjective life qualities and different smoking behaviors in order to determine the possible effects on tobacco use.

METHODSSix middle schools were chosen in the city and rural areas of Hefei in 1998 and 2003, respectively. Smoking behaviors such as ever smoking, smoking before 13 years old, current smoking, regular smoking and addictive smoking were surveyed. Emotional symptoms, satisfaction on school life and on general life were chosen to estimate subjective life qualities. Relationship between subjective life qualities and smoking behaviors was analyzed.

RESULTSPrevalence rates of smoking before 13 years old, current smoking, regular smoking and addictive smoking in 2003 (8.8%, 12.0%, 5.6% and 1.7%, respectively) were higher in 1998 (3.1%, 3.0%, 1.7% and 0.6%, respectively). The prevalence rates of depression and anxiety were high among students with ever smoking, current smoking, regular smoking, addictive smoking, smoking before 13 years old and no attempt to quit cigarette smoking. Lower school life satisfaction and general estimation of life were found among these students. Higher satisfaction on school life (OR = 0.657) and on ordinary life (OR = 0.766) were protecting factors of current smoking but comorbid depression and anxiety were risk factors (OR = 2.181). Higher school life satisfaction (OR = 0.388) seemed to be protecting factor of addictive smoking while depression was risk factor of addictive smoking (OR = 2.753) and regular smoking (OR = 1.676).

CONCLUSIONSmoking behaviors were common in middle school students and affected by several emotional and cognitive factors of subjective life qualities. Longitudinal designs are required to clarify the causal relationship between these factors and smoking behaviors in adolescents.

Adolescent ; Adolescent Behavior ; Anxiety ; epidemiology ; China ; epidemiology ; Depression ; epidemiology ; Health Surveys ; Humans ; Prevalence ; Psychology, Adolescent ; Quality of Life ; Risk Factors ; Rural Population ; statistics & numerical data ; Schools ; Smoking ; epidemiology ; Students ; psychology ; statistics & numerical data

Objective To investigate the episodes and influencing factors on self-harm and to explore the relationship between self-harm episodes and suicidal psychology and behaviors in college students. Methods Four universities were selected using cluster sampling method in Anqing city and Chaohu city. Totally, 2713 college students completed this survey. Data were analyzed by Pearson Chi-square and logistic regression. Results In the last six months, rates of highly lethal self-harm,less lethal self-harm with visible tissue damage, self-injury without visible tissue damage,self-harmful behaviors with latency damage, other self-harmful behaviors with menticide were 1.9%,5.5%, 15.3%, 21.2% and 17.0% respectively. The total rate of self-harm was 31.3%. 73.1% of the students with self harmful experiences had the above mentioned behaviors more than 3 times in the last six months. The top 3 reasons for taking self-harm actions were: having learning problems (43. 1% ), failed love affairs (25.0%) and having conflicts with others (23.9%). There were different influencing factors among different kinds of self-harm episodes. Depression was the risk factor of self-harm. The higher score of having high self-esteem was the protective factor of all kinds of self-harm actions except highly lethal ones. Higher score of difficulties in identifying feelings was one of the risk factors. The rates of suicidal psychology and behaviors in students with self-harm were significantly higher than those in students without those behaviors. Result from linear x2 test indicated that the graveness of tissue damage of self-harm was higher along with the rates of suicidal psychology and behaviors (P＜0.01). Conclusion Among 2713 college students, about 1/3adolescents having experienced self-harm in the last 6 months, many with repeated ones. Depression and difficulties in identifying feelings were the two risk factors while self-esteem was the protective factor related to most of the self-harm cases.

Objective This paper aimed to report the rate of suicide attempts among adolescent and young students of eight cities in China, and to examine the impact of related psychosocial factors on suicide attempts. Methods The investigation was obtained in senior and junior high school and college students in eight cities of China. All of the 17 622 participants were recruited to complete anonymous questionnaires regarding their experiences of suicide attempts and related psychosocial factors. Results Overall, 2.2 percent of the respondents reported that they had experienced attempted suicide during the previous year. Scores on high anxiety (OR=2.61,95%CI: 1.64-4.16),high depression (OR=2.05, 95%CI: 1.35-3.10),suicide idea (OR= 14.21, 95%CI: 9.88-20.43),suicide plan(OR=3.50,95%CI:2.59-4.73) were revealed as common risk factors while the whole time span being accompanied by mothers during juvenile years (OR=0.38, 95% CI: 0.20-0.74) seemed to be protective factor for suicide attempts through multivariate logistic regression analysis. Conclusion Our results showed that emotional symptom, idea and plans for suicide were statistically associated with suicide attempted in high school and college students.

Objective To examine the association between self-assessed somatotypes and depression related symptoms.Methods The study sample included 5555 students aged 9 to 18 years who had attended the Physical Fitness and Health Surveillance of Chinese School Students(2010)in Chizhou city,Anhui province.Association between self-assessed somatotypes,body mass index(BMI)and depression symptom were examined.Results There was a slight consistency between self-assessed somatotypes and BMI in both boys and girls(Kappa=0.217,P=0.000; Kappa=0.203,P=0.000).Significant difference in the prevalence of depression was found among weight misperception groups in both genders(x2=145.223,P=0.000).The prevalence of umderestimation of somatotypes was significantly higher in boys than in girls,while the result was reversive on the prevalence of overestimation.Additionally,the prevalence of depression was higher in girls than in boys(x2=5.199,P=0.023).Through logistic regression,data showed that self-assessed somatotypes and miscalculated groups were significantly associated with depressive symptoms.Compared to the group that self-assessment as being normal,those students when self-assessed as being slim,overweight or obesity were more likely to be depressive,with odds ratio(ORs)as 1.255(95％CI:1.066-1.478),1.538(95％CI:1.275-1.856)and 1.713(95％ CI:1.035-2.834),respectively.Overestimated and underestimated somatotypes appeared to be risk factors causing symptoms of depression(OR=1.705,95％CI:1.382-2.105; OR=1.241,95％CI:1.059-1.454).Conclusion Slight consistency was found between self-assessed somatotypes and BMI,while the misjudged somatotypes were the risk factor related to depressive symptoms.It was suggested that life skills education should be carried out as preventive intervention approach,to improve the physical and mental health well-being of children and adolescents.

Objective To investigate the association between psychological sub-healthy status and risks on injuries and related predictors among adolescents in China,in order to develop reasonable intervention programs.Methods Adolescents were selected from middle schools and colleges in eight cities.Baseline status on sub-healthy that related to psycho-pathology was measured using the Multidimensional Sub-health Questionnaire of Adolescents (MSQA).Data on Self- or unintentional injuries were collected using a standardized injury surveillance questionnaire every 3 months,for three times (T0,T1,T2 and T3).Results A total of 12 113 students completed the study.Prevalence rates related to self-injury among boys and girls were 19.4％ and 19.9％,respectively.The rates of having more than 4 types of self-injuries in boys and girls were 4.7％ and 4.2％,respectively.Boys had a significantly higher unintentional injury rate than that of girls (62.2％ vs.57.3％ ).The differences in the rates of having more than 4 types of unintentional injury between boys (3.8％) and girls (3.4％) were also statistically significant.Rates on self- and unintentional-injuries increased in T1,T2 and T3 when the psychological symptoms had an increase.Data from Multiple regression analysis demonstrated that the baseline psychological sub-healthy status was a predictive factor for higher risk of self- and unintentional- injuries at each of the follow-up waves.Conclusion Sub-healthy status of psycho-pathology was associated with an increased risk on self-and unintentional injuries among adolescents.Integrative psycho-pathological and behavioralintervention programs might help in reducing the risk of injuries among the Chinese adolescents.

Optimization of cultivation condition of recombinant E. coli DH5 alpha/pDH-B2m and the condition suitable for expression of recombinant mature peptide of human bone morphogenetic protein-2 was carried out in 500 mL shaking flasks and then transferred to NBS Bioflo IV, a 20 L DO feed-back fed-batch culture system, to obtain rhBMP-2. The results indicate that keeping dissolved oxygen at 40% and controlling nutrient feeding rate with DO feed back strategy can obtain theoretically 3.59 g recombinant mature peptide of hBMP-2 per liter of broth, the final cell density OD600 reaches 57(22.8 g dry cell weight/L), and the expression of rhBMP-2 amounts to 30% of the total protein in E. coli.
Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; biosynthesis ; genetics ; Culture Media ; Escherichia coli ; genetics ; Fermentation ; Glycerol ; pharmacology ; Recombinant Proteins ; biosynthesis ; Time Factors ; Transforming Growth Factor beta