Objective To investigate the transfection and expression of pcDNA3 1-VEGF 165 plasmid in mammalian cells. Methods pcDNA3 1-VEGF 165 plasmid was transfected into CHO cells by electroporation and the positive clones were screened by G418. The expression of VEGF 165 gene in the transfected CHO cells were detected by RT-PCR, ELISA and western blotting analysis. The biologic activities of VEGF 165 secreted by the transfected CHO cells were measured by Mile's assay and MTT method. Results G418-resistant CHO cell clones were obtained. RT-PCR, ELISA and western blotting analysis showed that there were the transcription and translation of VEGF 165 gene in anti-G418 CHO cells. The mile's assay showed that the expressed VEGF 165 could increase the vessel permeability, and MTT assay proved that it could promote the proliferation of endothelial cells. Conclusion pcDNA3 1-VEGF 165 plasmid could express VEGF 165 with biological activities in mammalian cells.

BACKGROUND: The occurrence and severity of acute high-altitude disease(AHAD) are determined by the speed entering the highland, the altitude of highland and seasons. The association between obesity and AHAD has not been fully investigated.OBJECTIVE: To investigate the association between obesity and acute high-altitude in people exposed rapidly to the highland.DESIGN: Comparative observation.SETTING: Department of Cardiology, General Hospital of Chinese PLA and Department of Cardiology, Tibetan General Hospital of Chinese PLA.PARTICIPANTS: The experiment was conducted at the Department of Cardiology, General Hospital of Tibetan Military Area Command of Chinese PLA in August 2006. Totally 82 Henan subjects aged 28-45 years were selected from the male workers constructing the Qinghai-Tibet Railway and their managers with acute high-altitude exposure. They had never been to the highland before, and all agreed to the detection.METHODS: ①Every subject completed the AHAD self-report questionnaire at sea level and 12 hours and 24 hours after ascending high-altitude. The items in the questionnaire included symptoms of headache, gastrointestinal symptoms,fatigue or weakness, dizziness, and insomnia. Each symptom was graded from 0-3 with 0 as no symptoms, 1 as mild symptoms, 2 as moderate symptoms, and 3 as severe symptoms and a total score of 15. A score of 4 or more could by identified as AHAD. ②The height and body mass were measured to calculate the body mass (BMI). Those with BMI≥28 kg/m2 served as the obesity group [n =39, mean age (35±8) years], and those with BMI of 18.5-23.9 kg/m2 as the normal body mass group [n =43, mean age (35±8) years]. ③Arterial blood was taken to evaluate arterial oxygen saturation (SO2), arterial oxygen pressure (PaO2) and arterial carbon dioxide pressure (PaCO2) at baseline and 24 hours after ascending high-altitude. ④The measurement data was compared by t test.MAIN OUTCOME MEASURES: BMI, vital capacity of lungs, SO2, PaO2 and PaCO2 levels of obese and normal people.RESULTS: Totally 39 obese people and 43 normal people were involved in the result analysis. ①AHAD score: No symptom was reported at sea level in all participants (scored 0), but the AHAD scores in the obesity group were significantly higher than those in normal group 12-hour and 24-hour after ascending high-altitude. ②Blood gas analysis:At sea level, there were no statistical differences in the levels of SO2, PaO2, and PaCO2 between two groups (P ＞ 0.05).But 24 hours after ascending high-altitude, SO2 and PaO2 of the obesity group were much lower than in the normal group (P＜ 0.01), and PaCO2 was significantly higher than in the normal group (P＜ 0.01).CONCLUSION: Obese men are more vulnerable to high-altitude hypoxia than people with normal body mass. Obesity is an important risk factor for the development of acute high-altitude disease.

Objective To construct the eukaryotic expression plasmid containing human epidermal growth factor(hEGF) gene with signal peptide(SP).Methods After two pairs of primers were designed and synthesized,the cDNA fragment of hEGF and SP genes were amplified from total RNAs. The amplified cDNA fragments were cloned into pGEM-T vector.The expression plasmids were verified by double endonuclease digestion and DNA sequence analysis. Results With RT-PCR using two pairs of primers,two bands(about 90bp and 180bp) were obtained and confirmed as signal peptide and EGF cDNA fragment with electrophoresis analysis and DNA sequencing after cloned into pGEM-T vector.The SP and EGF cDNA fragments were inserted into plasmid pcDNA3.1(+).The bands of 240bp and 5.4kb were obtained and identified as the full length of SP-EGF cDNA fragment by DNA sequence analysis.Conclusion The eukaryotic expression plasmids containing hEGF gene is successfully constructed.

OBJECTIVEAn accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.

METHODScopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.

RESULTGood linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.

CONCLUSIONThe method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.

China ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Convolvulaceae ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Scopoletin ; analysis

Objective To evaluate the treatment effects of middle and proximal phalangeal fractures with lateral fixation of mini-titanium plate.Methods From December 2011 to April 2015,56 patients (64 phalanges) suffered from closed middle and proximal phalangeal fractures were treated with lateral fixation of mini-titanium plate.There were 48 males and 8 females,aged from 17 to 65 years (average,36.3 years).Fractures included 25 cases of middle phalangeal fracture and 39 cases of proximal phalangeal fracture.According to site of fracture,12 cases were middle phalangeal condyle fractur,7 cases were middle phalangeal shaft fracture,6 cases were middle phalangeal base fracture,10 cases were proximal phalangeal condyle fracture,22 cases were proximal phalangeaal shaft fracture and 7 cases were proximal phalangeal base fracture.According to the type of fracture,13 cases were transverse,9 cases were short oblique,11 cases were long oblique,6 cases were spiral and 25 cases were comminuted fracture.All phalangeal fractures were exposed by lateral approach,in which the lateral band and oblique fibers of proximal phalanx were excised to fully expose proximal phalangeal fracture.After the reduction,the fractures were fixed with mini-titanium plate laterally.The Disabilities of the Arm,Shoulder and Hand (DASH)and the bilateral fingers total active motion (TAM) were employed to evaluate the functions.All patients took X ray during follow-up.Results All 56 patients were followed up from 9 to 47 months,average (14 ± 6) months.Phalangeal fractures were healed from 7 to 14 weeks,average (8.8 ± 2.4) weeks without loss of fixation or malunion.At final follow-up,DASH score were from 1.7 to 7.5,with an average of 4.8 ± 2.2.TAM of the fingers was excellent (＞ 90％ TAM of the contralateral side) in 33/64 (51.5％) cases,good (75％-90％ TAM of the contralateral side) in 30/64 (46.9％) cas es,fair (50％-75％ TAM of the contralateral side) 1/64 (1.6％) cases.Conclusion Lateral approach fully exposes middle and proximal phalangeal fractures and less interferes with the extensor mechanism.Lateral fixation with mini-titanium plate could provide stability and allow early motion.This technique may be an optional choice in clinical practice.

Flexible matching has recently been proposed as a method of improving interactions efficiency. In this study, the concept of flexible matching has been introduced, and the applicability of this strategy has also been described based on the power calculation of interaction between HER-2 polymorphism and smoking with breast cancer. A large-sample approximation method is used to estimate the power and efficiency of gene-environment interactions. In the basic scenario, power of interaction between HER-2 polymorphism and smoking of unmatched case-control study appears to be 30% while in the frequency matching case-control study it is 56%. However, when increasing the smoking prevalence in controls, greater power can be obtained (power=74%). Conclusions: Flexible matching strategies can increase the power and efficiency of case-control studies to detect and estimate the gene-environment interactions when compared with traditional frequency matching and it is especially useful under those scenarios when low environmental exposure of population, adverse gene-environment interactions or less paired controls are seen. Optimal matching design should be made available by weighing the benefits and loss due to flexible matching.

Objective:To explore influence of thrombus aspiration combined bivalirudin during emergency PCI on my- ocardial tissue perfusion and clinical prognosis in patients with acute ST elevation myocardial infarction (STEMI). Methods:A total of 102 patients with acute STEMI,who were confirmed with thrombus burden by CAG in our hos- pital from Jan 2012 to Jun 2014,were selected.According to random number table,they were randomly divided into thrombus aspiration + bivalirudin group (n=52,thrombus aspiration group)and heparin group (n=50,routine PCI group).TIMI blood flow grade 3 rate,TIMI myocardial perfusion grade (TMPG)after PCI,ST segment re- gression rate 2h after PCI,peak value and peak time of cTnI after PCI,LVEF,LVEDd,incidence rates of bleeding and major adverse cardiovascular events (MACE)on one week and one month after PCI were compared between two groups.Results:Compared with routine PCI group,there were significant rise in postoperative TMPG grade 3 rate (56.00% vs.88.46%),TIMI grade 3 rate (58.00% vs.88.46%)and ST segment regression rate (52.00% vs. 76.92%)in thrombus aspiration group,P<0.05 or <0.01. Compared with routine PCI group one month after PCI,there was significant rise in LVEF [(53.76±5.24)% vs.(57.95±5.51)%],and significant reductions in LVEDd [(53.70±3.39)mm vs.(50.63±1.24)mm],peak value [(16.00±4.28)μg/L vs.(13.81±4.00)μg/L]and peak time [(14.00±2.80)h vs.(13.00±2.23)h]of cTnI in thrombus aspiration group,P<0.05 or <0.01. Incidence rate of mild bleeding in thrombus aspiration group was significantly lower than that of routine PCI group (1.9% vs.16.0%),P<0.05,but there was no significant difference in incidence rate of MACE between two groups,P>0.05. Conclusion:Thrombus aspiration combined bivalirudin during emergency PCI is safe and fea- sible for acute STEMI patients,it can effectively reduce incidence rate of bleeding,remove coronary thrombus,im- prove myocardial tissue perfusion and doesn't increase incidence rate of MACE.

The reform and practice on teaching approaches of microbiology integrated case-based study,heuristic teaching,interaction teaching,comparison expression teaching and inductive learning was explored,which was contraposed the education aims of independent college and student's trait,and combined to the fact of University of Electronic Science and Technology of China Zhongshan Institute.The result of practice showed that student enthusiasm for study was mobilized,and better results in teaching were got by applica-tion of diversiform teaching approaches.

AIM: To report one case of huge fungal granuloma in the deep maxillofacial region with its clinical findings,diagnosis and treatment process to improve our understanding of fungal granuloma and lessen misdiagnosis.METHODS: The literature was reviewed with combination of case report.RESULTS: The clinical and imaging findings of fungal granuloma had no specificity and were very subject to misdiagnosis. Confirmed diagnosis needed histopathological examination and bacterial culture.CONCLUSION: Deepened understanding of this disease can decrease misdiagnosis and benefit prompt and proper treatment.

OBJECTIVE: To investigate the expression of monocyte chemotactic protein-1 (MCP-1) and its receptor CC chemokine receptor-2 (CCR-2) in coronary atherosclerosis plaques between sidden coronary death (SCD) and non-SCD. Methods The expression levels of MCP-1 and CCR-2 in SCD group, coronary atherosclerosis group (non-SCD), control group (normal coronary artery) were detected by immunohistochemistry. RESULTS: Positive rates of MCP-1 among the three groups were 78%, 47%, and 0%, respectively, with significant expressing differences between each two groups (P<0.05). Positive rates of CCR-2 among three groups were 72%, 47%, and 0%, respectively, with significant expressing differences between the SCD group and coronary atherosclerosis group as well as between the SCD group and control group (P<0.05), but with no significant expressing difference between coronary atherosclerosis group and control group (P>0.05). CONCLUSION Overexpression of MCP-1 and CCR-2 in coronary atherosclerotic plaques is closely correlated with SCD.
Chemokine CCL2/metabolism* ; Coronary Artery Disease/pathology* ; Death, Sudden, Cardiac/pathology* ; Humans ; Immunohistochemistry ; Receptors, CCR2/metabolism*