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.

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

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 provide recommendations for the management of intensive care unit patients without novel coronavirus disease 2019 (COVID-19). Methods    We set up a focus group urgently and identified five key clinical issues through discussion. Total 23 databases or websites including PubMed, National Guideline Clearing-House, Chinese Center for Disease Control and Prevention and so on were searched from construction of the library until February 28, 2020. After group discussion and collecting information, we used GRADE system to classify the evidence and give recommendations. Then we apply the recommendations to manage pediatric intensive care unit in the department of  critical care medicine in our hospital. Results    We searched 13 321 articles and finally identified 21 liteteratures. We discussed twice, and five recommendations were proposed: (1) Patients should wear medical surgical masks; (2) Family members are not allowed to visit the ward and video visitation are used; (3) It doesn’t need to increase the frequency of environmental disinfection; (4) We should provide proper health education about the disease to non-medical staff (workers, cleaners); (5) Medical staff do not need wear protective clothing. We used these recommendations in intensive care unit management for 35 days and there was no novel coronavirus infection in patients, medical staff or non-medical staff. Conclusion    The use of evidence-based medicine for emergency recommendation is helpful for the scientific and efficient management of wards, and is also suitable for the management of general intensive care units in emergent public health events.

Humans ; Pilot Projects ; Prospective Studies ; Pulmonary Emphysema/surgery* ; Lung/surgery* ; Emphysema ; Perfusion ; Tomography, Emission-Computed ; Tomography ; Tomography, Emission-Computed, Single-Photon ; Pneumonectomy ; Bronchoscopy/methods*