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

Objective To investigate the CD44v6 and heparanase in non-small cell lung cancer (NSCLC ) and adjacent normal lung tissues ,and to study the correlation of these two molecular in non-small cell lung cancer invasion and metastasis .Methods The expressing of CD44v6 and heparanase in 117 patients with non-small cell lung cancer tissues and 62 adjacent normal lung tissue was detected by immunohistochemical staining .Results The expression of CD44v6 in NSCLC was 54 .7% (64/117) ,which was sig-nificantly higher than in adjacent tissues of 4 .8% (3/62)(P<0 .05);the expression of heparanase in NSCLC was 86 .3% (101/117) ,which was significantly higher than the normal adjacent tissues 17 .7% (11/62)(P<0 .01);CD44v6 expression in NSCLC ex-pression was positive correlated with heparanase .Conclusion CD44v6 and heparanase are associated with the invasion and metasta-sis of NSCLC ,combined detection of the expression of the two molecular maybe has a certain reference value in determine the prog-nosis of NSCLC .

Objective To compare the clinical outcome of external fixation device with Kirschner wire or fragment fixation pin with mini plate and screw fixation in treatment of metacarpal and phalangeal articular fracture dislocation. Methods From October 2002 to March 2008, 106 patients with metacarpal and phalangeal articular fracture dislocation were randomly divided into A and B group. The 53 patients in A group were treated with external fixation device with Kirschner wire or fragment fixation pin. There were thumb injury in 24 cases, fracture-dislocation of proximal interphalangeal joint(PIP) in 36 cases, fracture-dislocation of metacarpophalangeal joint (MP) in 17 cases. The 53 patients in B group were treated with mini plate and screw fixation. There were thumb injury in 22 cases, fracture-dislocation of PIP in 30 cases, fracture-dislocation of MP in 23 cases. Duncan rating criteria were used to compare finger range of motion. Results The mean follow-up of 16.8 and 17.5 months in A group and B group. According to Duncan rating criteria, there were excellent in 33 cases, good in 16, fair in 3, and poor in 1 case. The excellent and good rate was 92.5% in A group. There were excellent in 30 cases, good in 17, fair in 5, and poor in 1 case. The excellent and good rate was 88.7% in B group. In A group, 1 case of wound infection was found. The average arc of motion of thumb joint was 134°±21° while the average arc of motion of other fingers was 248°±19°. In B group, no wound infection occurred. The average arc of motion of thumb joint was 122°±18° while the average arc of motion of other fingers was 225°±17°. Conclusion External fixation device with Kirschner wire or fragment fixation pin was better than the application of mini plate and screw fixation in treatment of metacarpal and phalangeal articular fracture-dislocation.

Objective To investigate the effectiveness and safety of low-intensity warfarin anticoagulation in over 80-year-old patients with nonvascular atrial fibrillation (NVAF). Methods The 180 NVAF patients aged over 80 years were randomly assigned into 2 groups: 90 patients in lowintensity warfarin anticoagulation group (target value of INR 1.6-2.0), the other 90 patients in standard-intensity warfarin anticoagulation group (target value of INR 2. 0-3.0). All patients were followed up in outpatient-department for one year. Main outcome measures included the incidence rates of bleeding and thromboembolic events, and secondary outcome measures included the warfarin dosage and times of INR＞3.0. Results The incidence rate of thromboembolic events was 4.4% (4/90) in low-intensity group and 3.3% (3/90) in standard-intensity group with no statistically significant difference between these two groups (P＞0. 05). However, the incidence rate of hemorrhage was significantly lower in low-intensity group than in standard-intensity group [5.6% (5/90) vs. 16.7%(15/90), P＜0. 05]. Meanwhile the warfarin dosage was significantly lower in low-intensity group than in standard-intensity group [(1. 55±0. 63) mg vs. (2.31±0.57) mg, P＜0.05]. The times of INR＞3.0 were less in low-intensity group than in standard-intensity group (P＜0. 05). Conclusions Therapy with low-intensity warfarin anticoagulation in NVAF patients aged over 80 years may be equally effective as, but safer than that with standard-intensity warfarin.

Background Studies show that regulatory T cells (Treg) are a kind of T cell subsets to negatively regulate immune response,and play an important role in maintaining immune homeostasis and immune tolerance.Autoimmune uveitis is an autoimmune disease,the regulation of Treg cells in pathogenesis and progression of autoimmune uveitis is not fully unelucidated.Objective This study was to observe the dynamic changes of Treg in experimental autoimmune uveitis (EAU) rats and explore the role of Treg cells in the pathological process of EAUrats.Methods Eighty four 6-8 week-old SPF Lewis rats were randomly divided into model group and control group.The mixed emulsifier of interphotoreceptor retinoid-binding protein (IRBP)1177-1191,tuberculin (TB),complete Freund adjuvant (CFA) and PBS (300 μl) was subcutaneously injected in double rear foot pad,abdominal side and back,and only equal amount of TB,CFA and PBS emulsifier was used in the same way in the control group.Ocular inflammation symptoms was examined at 9,13,18,23,28,35 and 48 days after modeling and scored based on the severity of the inflammatory.Six rats of each group were sacrificed in above time points respectively for the histopathological examination of iris,ciliary body and retinas by haematoxylin-eosin staining.The lymphocytes were isolated and cultured from rat spleens,and the proportion of Foxp3-labelled cells,a specific marker of Treg cells,was assayed by flow cytometry.The relative expression level of Foxp3 mRNA in the lymphocytes detected by using realtime quantitative PCR (RT-PCR).The use and care of the rats complied with the ARVO Statement.Results Eye inflammatory response appeared at 8 days after immunization,showing vasodilation and hyperemia of rat iris in the model group,and the response peaked at 13 days,with exudation and hypopyon in the anterior chamber.The highest inflammatory scores were 3.75±0.42 at day 13,and the ocular inflammation reaction was gradually relieved after that and disappeared at 23 days after immunity.A significant difference in ocular inflammatory scores of model rats was found among different time points (F =81.709,P ＜ 0.001);while no inflammatory symptom was observed in the control group.Histopathology examination showed obvious infiltration of inflammatory cells in the iris,ciliary body and retinas in model rats,including neutrophils,lymphocytes and mononuclear cells.The proportion of Foxp3-labelled cells in spleen lymphocytes was (5.50 ± 0.64)％,(13.36 ± 0.98)％,(10.34 ± 0.79)％,(9.58 ± 1.02)％,(6.73 ±0.81)％ and (5.58 ± 0.47) ％ in the model group on day 13,18,23,28,35,48 respectively,with statistically significant differences in comparison with (2.80 ± 0.38) ％,(3.36 ± 0.53) ％,(3.65 ± 0.57) ％,(3.37 ± 0.43) ％,(3.33±0.50)％ and (3.13±0.61)％ in the control group (t=-6.272,-15.556,-11.910,-9.753,-6.154,-5.491,all at P＜0.01).The change trend of Foxp3 mRNA expression was consistent to the dynamic change of the proportion of Foxp3-labelled cells.Conclusions The pathogenesis and development is closely associated with the dynamic changes of CD4+ CD25 + Foxp3+ Treg cells in EAU rats.

Objective To investigate effect of cluster nursing for children with severe pneumonia in ICU.Methods A total of 88 patients with severe pneumonia were randomly divided into two groups,the control group received routine care,and the experimental group conducted cluster nursing.Blood gas analysis results after intervention,adjuvant effect of nursing,related clinical indicators,VAP,mortality and nursing satisfaction were compared.Results The experimental group had higher levels of p (O2),SaO2 and pH,lower p (CO2) level,ventilation time,ICU time and length of hospital stay and incidence of VAP than the control group (P ＜ 0.01).The experimental group had higher total efficiency and nursing satisfaction,and lower mortality than the control group (P ＜ 0.05).Conclusion Cluster nursing for children with severe pneumonia in ICU has significant efficacy,and can significantly improve nursing satisfaction.

Objective To discuss the influence of water colloid dressing on the skin of tracheal intubation in less than 6 months newborns with congenital heart disease.Methods A total of 80 newborns with congenital heart disease accepted therapy in our hospital were selected and were divided into control group and experimental group,with 40 cases in each group.3 M elastic tape was used to fix tracheal intubation in control group while water colloid dressing was used to protect the cheek and then 3 M elastic tape was applied to fix catheter in experimental group.The skin condition of the cheek was compared after intubation.Results The skin condition of experimental group was significantly better than control group (P ＜ 0.05).Conclusion Water colloid dressing could protect the skin and decrease the rate of avulsion injury of skin and pressure sores.

Objective To investigate effect of cluster nursing for children with severe pneumonia in ICU.Methods A total of 88 patients with severe pneumonia were randomly divided into two groups,the control group received routine care,and the experimental group conducted cluster nursing.Blood gas analysis results after intervention,adjuvant effect of nursing,related clinical indicators,VAP,mortality and nursing satisfaction were compared.Results The experimental group had higher levels of p (O2),SaO2 and pH,lower p (CO2) level,ventilation time,ICU time and length of hospital stay and incidence of VAP than the control group (P ＜ 0.01).The experimental group had higher total efficiency and nursing satisfaction,and lower mortality than the control group (P ＜ 0.05).Conclusion Cluster nursing for children with severe pneumonia in ICU has significant efficacy,and can significantly improve nursing satisfaction.

Objective To discuss the influence of water colloid dressing on the skin of tracheal intubation in less than 6 months newborns with congenital heart disease.Methods A total of 80 newborns with congenital heart disease accepted therapy in our hospital were selected and were divided into control group and experimental group,with 40 cases in each group.3 M elastic tape was used to fix tracheal intubation in control group while water colloid dressing was used to protect the cheek and then 3 M elastic tape was applied to fix catheter in experimental group.The skin condition of the cheek was compared after intubation.Results The skin condition of experimental group was significantly better than control group (P ＜ 0.05).Conclusion Water colloid dressing could protect the skin and decrease the rate of avulsion injury of skin and pressure sores.

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