This study aims to construct the plasmid of human Runx1 and observe its possible effects on Runx1 gene expression in human pulmonary adenocarcinoma cells (A549). The shRNA sequence targeting human Runx1 was designed and synthesized, then inserted into pSuper plasmid by DNA recombination technology. The recombinant plasmid was confirmed by bacterial colonies PCR, enzyme digestion analysis and DNA sequencing. A549 cells were transfected by Runx1 shRNA plasmid. The inhibition efficiency of pSuper-Runx1-shRNA plasmid on Runx1 at mRNA level and protein level were measured with real-time PCR and Western blot. The results of real-time PCR and Western blot indicated that the mRNA and protein levels of Runx1 in A549 cells were inhibited by the pSuper-Runx1- shRNA expression plasmid, and the inhibition rate were 33% and 50%, respectively.
Cell Line, Tumor ; Core Binding Factor Alpha 2 Subunit ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Transfection

Objection To explore adenylate content of traumatic brain edema tissue and the relationship between ad-enylate content and brain edema.Methods The 20 rabbits with traumatic brain edema were sacrificed by decapitation in different periods,other 4 rabbits brain as control group.The brain tissue in the ringed traumatic site was used to measure brain water content and adenylate including AMP,ADP and ATP contents with HPLC.Results The adenylate content of traumatic brain edema tissue was lower than that of normal brain tissue,and further obviously reduction with longitude after trauma,and consistent with brain edema extent.Condusion The adenylate content of traumatic brain tissue is reduced along with brain edema severity and there is obviously correlation.

Objective To exam the personality traits and self-concept of school phobia patients with different academic achievements.Methods 38 school phobia patients with high academic achievement and 19 school phobia patients with low academic achievement were assessed with Eysenck' s personality questionnaire and Piers-Harris Children' s Self-concept Scale.Results High academic achievement group of psychoticism and neuroticism higher than that of low academic achievement group.High academic achievement group of Self-concept's total scores,behavior,intelligence and the school situation higher than that of low academic achievement group.High academic achievement group of anxiety scale score lower than that of low academic achievement group.The personality traits of school phobia correlated with the self-concept on some level.Conclusion Different academic achievements of school phobia patients have different personality traits and have different self-concept,so they need to individualized treatment options.

AIM: To investigate the antioxidant status and the oxidative damage of cellular macromolecules in patients with Graves' disease. METHODS: Fasting plasma level of total antioxidant capacity (TAC), and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured in 31 patients with untreated Graves' disease and 31 treated Graves' disease. DNA damage of peripheral blood mononuclear cell (PBMC) was detected by single cell gel electrophoresis assay (expressed in comet percentage). In addition, thiol group (SH) and malondialdehyde (MDA) were measured. 31 age-matched healthy subjects were studied as a control group. RESULTS: Plasma TAC, SOD and GSH-Px were significantly lower in patients with untreated Graves' disease compared to the controls (P

The aim of this study was to observe whether necroptosis is involved in the process of cardiac hypertrophy induced by pressure overload. SD rats underwent transverse abdominal aortic constriction (TAC) operation for establishing cardiac hypertrophy model. The structure and function of the left ventricle of rats were evaluated via echocardiography, left ventricular mass index, the expression of markers of cardiac hypertrophy and histological detection. Real-time PCR and Western blot were used to measure the gene and protein expression of receptor interacting protein kinase 1 and 3 (RIPK1 and RIPK3, the necroptosis markers) respectively. Four weeks after TAC operation, rat model for cardiac hypertrophy was established. The experimental data showed that the gene and protein expressions of RIPK1 and RIPK3 in the rat heart hypertrophic tissues after TAC for 4 weeks were increased significantly compared with those in the sham group. HE staining showed cardiomyocytes injury and hypertrophy in the hearts of TAC rat models. By transmission electron microscope, we observed that mitochondria of cardiomyocytes were damaged seriously in the TAC models. Treatment with losartan used, the selective antagonist of angiotensin II type I receptor could improve the cardiac function of TAC rats. Moreover, losartan treatment decreased the expression of RIPK1 and RIPK3 in heart tissues of TAC rats. The results suggest that necroptosis occurrs in the process of cardiac hypertrophy with pressure overload, and losartan could alleviate the cardiac hypertrophy and inhibit necroptosis.
Animals ; Apoptosis ; Cardiomegaly ; pathology ; Disease Models, Animal ; Echocardiography ; Heart ; physiopathology ; Losartan ; pharmacology ; Myocytes, Cardiac ; Pressure ; Protein-Serine-Threonine Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; metabolism

For investigating the effect of Jumonji domain-containing protein-3 (JMJD3) on the behavior of lung cancer cell line, A549 proliferation was measured with EDU staining and flow cytometer after JMJD3 expression plasmid and pcDNA3. 1 transfection at 48h. The migration ability of A549 was tested at the same time. The expression of p21 mRNA was measured with RT-PCR. The results showed that JMJD3 transfection increased the EDU positive cells ratio (JMJD3: 40.75% +/- 2.07%, control: 20.97% +/- 1.5%, P < 0.001). G1 phase cell ration also increased after JMJD3 transfection (JMJD3:47. 80% +/- 1.85%, control: 54.60% +/- 0.95%, P = 0.005). The mRNA expression of p21 decreased in JMJD3 group (JMJD3: 35. 89% +/- 3.71%, control: 91.78% +/- 3.74%, P < 0.001). The distances of migration were (0.47 +/- 0.27) cm and (0.96 +/- 0.40) cm after 24h and 48h with JMJD3 tranfection, compared to (0.57 +/- 0.22)cm and (1.08 +/- 0.33)cm in control, respectively (P > 0.05). JMJD3 promoted the proliferation of A549 and decreased the G1 cell numbers, decreased the p21 mRNA, but had no effect on A549 migration.
Adenocarcinoma ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Humans ; Jumonji Domain-Containing Histone Demethylases ; pharmacology ; Lung Neoplasms ; pathology ; RNA, Messenger ; genetics ; metabolism ; Transfection

microRNAs (miRNAs) are a family of important small non-coding RNA molecules, which participate in the post transcriptional gene regulation. In this review, the numbers and chromosomal distribution of chicken miRNAs, and the regulation and function of chicken miRNAs in immune, embryo development and virus infection were reviewed. Additionally, the applications of chicken miRNAs were also discussed briefly. We hope it can provide references for further study and use of miRNAs in poultry husbandry fields.
Animals ; Chickens ; genetics ; Embryonic Development ; genetics ; Immunity ; genetics ; MicroRNAs ; genetics ; Virus Diseases ; immunology ; veterinary

Objective To study the inducing factors and clinical characteristics of patients hospitalized for asthma exacerbation in China. Methods Patients hospitalized for asthma exacerbation at 29 hospitals in China were retrospectively recruited during 2013-2014. Results Clinical data of 3 240 asthmatic patients were collected and analyzed including 1 369(42.3%) males and 1 871(57.7%)females. The patients hospitalized for asthma exacerbation counted for 2.95% (6 375/215 955) of all patients hospitalized during the same period. The leading six inducing factors, in sequence, were acute upper respiratory tract infection[42.3%(1 370/3 240)],changes of weather[22.8%(738/3 240)],noxious gas[(4.3% (140/3 240), allergy challenges [3.5%(115/3 240)], strenuous exercise [1.8%(57/3 240)], and air pollution [1.5%(49/3 240)].In older patients,more exacerbations were induced by weather changes,yet less sensitive to allergy challenges. As to middle-aged patients, they were less sensitive to upper respiratory tract infections,however the difference was not statistically significant(P>0.05).In winter more asthma patients were induced by upper respiratory tract infections,while in autumn more patients were induced by weather changes,strenuous exercise and air pollution.In spring and summer more patients were induced by allergy challenges, but the differences failed to achieve statistical significance (P>0.05). In northern cities more patients were induced by upper respiratory infections, whereas in southern cities more by noxious gases. Allergy challenges and air pollution tended to affect more patients in northern cities,but the difference was of no significance (P>0.05). The differences of inducing factors among patients of different gender, with or without a smoking history, and with different exacerbation severity didn't show any statistical significance. The patients with severe and life-threatening exacerbations counted for 20.1%(652/3 240).The percentage of patients older than 60 years was higher in patients with severe or life-threatening exacerbations than in whose with mild or moderate exacerbations,so did the percentage of male patients,of patients with disease duration longer than 10 years, with smoking history, and with a history of hospitalization or emergency department visits due to asthma exacerbation during the last year.Conclusion The acute upper respiratory tract infection ranks top among all the inducing factors. Senility, male gender, long duration of disease, smoking history, and a history of frequent hospital visits might be the risk factors for severe or life-threatening asthma exacerbations.

This study was aimed to construct transgenic mouse model with target for Runxl gene. Runxl cDNA of mice was amplified by PCR from pcDNA3. 1 Flag Runx1 FL vector and inserted into ptetO7-Asc-IRES-EGFP vector to form a recombinant vector, and then the recombinant vector was injected into fertilized egg by microinjection technology to get a transgenic mouse. The results of PCR and Southern blot indicated that the Runx1 transgenic mouse was constructed successfully, and this could provide an important tool for studying the function of Runxl gene in vivo.
Animals ; Base Sequence ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Genetic Vectors ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; genetics ; Microinjections ; Molecular Sequence Data ; Recombination, Genetic

Objective:To explore the clinical effects of a single stage reconstruction and revascularization using a free Flow-through chimeric anterolateral thigh perforator (ALTP) flap in Gustillo III C limb injuries.Methods:From January, 2010 to December, 2017, 17 patients with Gustillo III C injury of extremities were repaired with Flow-through chimeric ALTP in emergency surgeries. The patients were 16 males and 1 female with mean age of 32.4 (19-55) years. The size of wounds ranged from 16 cm×8 cm-45 cm×30 cm. The injured arteries were Flow-through anastomosed with the descending branch of the lateral circumflex femoral artery to regain blood flow. The deep dead space was filled with vastus lateralis flap, and the skin flap and fascia flap were used for superficial covering. The donor site was closed directly in 6 patients, simultaneous skin grafts were applied in 11 patients.Results:The followed-up time ranged 5 to 60 (average 21.8) months, and conducted by clinic visits and by telephone or WeChat interviews. Fifteen flaps survived, and 2 flaps failed with limb amputated. Six patients were repaired with skin and fascia flaps and 11 patients received flaps comprising the skin, fascia and vastus lateralis muscle. Four flaps healed in stage I. Partial necrosis were observed in 11 flaps. Of which, 1 healed by change of dressing and 10 healed by skin grafting. No complications occurred in donor site in all patients.Conclusion:The Flow-through chimeric ALTP flap can be used in stage I reconstruction of the blood supply of limbs and meanwhile achieve the 3-D repair of defects. It is one of the reliable methods in the reconstruction of Gustillo III C injuries in the extremities.