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 By the modern manufacture regulation of original equipment manufacture(OEM),the quality of large scale X-ray equipment is ensured by means of assembling the purchased key components,such as high frequency and high voltage generators and X-TV systems,etc.Methods The technology of Programmable Logic Controller(PLC)was utilized to improve the reliability of X-ray examination table.Results The design of PLC-controlled X-ray examination table was used in the digital X-ray equipment yeas ago.The malfunction probability of mechanic parts was very low,whereas the electric circuit part was almost zero.Conclusion The practice of PLC improves the reliability and control level for X-ray equipment.

Objective To improve X-ray equipment for better medical treatment by digitally transform normal X-ray equipment.Methods Digital image workstation was equipped for digital photography so that ordinary X-ray equipment has the function of a digital X-ray equipment.Results The digital transformation enhance the clinical performance of X-ray equipment.Conclusion It has been proved that some ordinary X-ray equipment can be equipped with digital workstation.

BACKGROUND: Using low-intensity pulsed ultrasound (LIPU) to promote the repair of articular cartilage injury is very common,and we also have more options to choose the cytoskeleton, but the application conditions of LIPU and the appropriate cytoskeleton have not reached any consensus yet.OBJECTIVE: To investigate the feasibility of establishing tissue-engineered cartilage by cell-free allograft demineralized bone matrix (CFDBM) co-cultured with rabbit cartilage cells and bone marrow mesenchymal stem cells (BMSCs) in vitro, and to investigate the effect of LIPU on the cells in CFDBM.DESIGN, TIME AND SETTING: Multiple sample observation was performed at the Institute of Biomedical Engineering, Second Military Medical University of Chinese PLAfrom May to August 2009.MATERIALS: The CFDBM was prepared as modified Urist's method; the cartilage cells were obtained using mechanical disintegration and enzyme digestion; the BMSCs were separated using whole bone marrow rinsing method, purified, and amplified layer by layer.METHODS: As CFDBM With a composite of different cellular components, and whether applying LIPU stimulation, the samples were divided into four groups: chondrocyte group, BMSCs group, compound group (CFDBM was compounded with chondrocytes,BMSCs, and chondrocytes/BMSCs, respectively, without LIPU stimulation), and LIPU group (CFDBM was compounded with chondrocytes/BMSCs, and then the samples were stimulated with LIPU on the second day, 1.0 MHz frequency, 10 mW/cm~2 transient spatial intension, 20 min/d).MAIN OUTCOME MEASURES: ① the 2~(nd)-generation of cartilage cells and BMSCs were examined by immunohistochemical method; ② The CFDBM prepared as modified Urist's method was examined as HE staining; ③ The samples of four groups were examined by collagen II immunohistochemical staining on the 21~(st) day.RESULTS: ① The collagen II immunohistochemical staining of the second generation of the articular cartilage cells showed that the morphostructure was polygon, star or round, and pseudopodia extended, and the cells were rich in cytoplasm; the cytoplasm was brownish yellow, and the cell nuclear was round. ② The result of immunohistochemical staining of BMSCs showed that,CD34 was negative, CD44 and CD105 were positive. ③ In the center of CFDBM prepared as modified Urist's method, there was no obvious cell-like structure and the gap size was uniform. ④ On the 21~(st) day after combining CFDBM with cells, collagen II immunohistochemical staining demonstrated that BMSCs group was negative, chondrocyte group was weak positive, compoundgroup was positive, and the LIPU group was strongly positive.CONCLUSION: ① Biological property of the 1~(st)-3~(rd)-passage chondrocytes and BMSCs was similar to primary-cultured cells. ②Both chondrocytes and BMSCs had a highly proliferative ability in CFDBM. ③ 10 mW/cm~2 LIPU could not affect activity of BMSCs but could promote differentiation Into articular cartilage cells, and it also could not promote celt proliferation.

OBJECTIVE: To study the effect of total flavonoids from Alpinia officinarum on gastrointestinal motility.METHODS: Gastric emptying method was applied to observe the effect of total flavonoids from A.officinarum on gastric emptying of normal mice and sthenic gastric emptying induced by pyridostigmine bromide.Effect of total flavonoids from A.officinarum on gastric smooth muscle of rats was observed through in vitro test and small intestine advancement was used to observe the influence of total flavonoids from A.officinarum on intestinal motility.RESULTS: Total flavonoids from A.officinarum had no significant influence on gastric emptying of normal mice,but distinctly inhibited sthenic gastric emptying induced by pyridostigmine bromide(P

OBJECTIVE:To investigate the perioperative use of antibiotics in patients undergoing colorectal operations. METHODS:The medical records of 213 patients undergoing colorectal operations in 2006 were randomly selected from the gastrointestinal surgery department of our hospital and divided into type Ⅰ operation group and type Ⅱ operation group accord-ing to patients’ operation time and wound contamination.The kinds and medication duration of antibiotics used before,during and after operation were investigated,and the postoperative infections were analyzed.RESULTS:The preoperative preventive medication were all conducted within the first 1～3days,and the dosage schemes were unified with aminoglycosides and metronidazole topping the list.The antibiotics used during operation were all in single dose,with 3rd generation cephalosporins combined metronidazole predominantly used.Postoperative use of antibiotics was somewhat complicated:type Ⅰ operation patients were treated more often by 2 kinds of antibiotics concurrently for 3～7days and type Ⅱ operation patients were treated more often by 3 kinds of antibiotics concurrently for at least 6～7days.The incidence of postoperative infection was 3.4% in type Ⅰ operation patients as compared to 11.6% in type Ⅱ operation patients,showing significant differences between two groups.CONCLUSION:The perioperative application of antibiotics in patients undergoing colorectal operation was reasonable on the whole,with 3rd generation cephalosporins plus metronidazole predominantly used,but there were some problems in the postoperative medication.

Objective To investigate the risk factors and the influence of intravenous thrombolysis of acute cerebral infarction with cerebral microbleeds(CMBs)by SWI.Methods 1 64 patients with acute cerebral infarction were enrolled in this study.All pa-tients were scanned with routine MRI and SWI.According to the presence of CMBs on SWI,the patients were classified into two groups:CMBs group(73 cases)and non-CMBs group(91 cases).Past history was recorded and risk factors of CMBs were explored. 76 cases patients(including 35 cases of CMBs group and 41 cases of non-CMBs group)were treated by intravenous thrombolysis and rescanned with routine MRI and SWI to compare the changes in the number of CMBs and hemorrhage transformation 24 hours after thrombolysis.Results The difference age,hypertension,lacunar infarction and leukoaraiosis between the two groups were significant (P <0.05).The difference of CMBs and hemorrhagic transformation between the two groups treated by thrombolysis were not sig-nificant(P >0.05).Conclusion Acute cerebral infarction with CMBs are influenced by age,hypertension,lacunar infarction and leu-koaraiosis.Thrombolysis in acute cerebral infarction with CMBs can not augment the incidence of hemorrhagic transformation.

Objective To explore the feasibility of establishing a swine model of liver cirrhosis with portal hypertension by portal infusion of 80％ alcohol.Methods A total of 13 Guizhou miniature pigs were randomly divided into three groups,experiment group 1 (n=5),experiment group 2 (n=5) and control group (n=3).Experiment groups of pigs received portal infusion of 80％ alcohol in volumes of 5 ml in group 1,and 10 ml in group 2,and the pigs in control group received portal perfusion of saline in volumes of 10 ml.All animals were performed direct portal angiography,the portal vein pressures and diameter were also detected before,immediately and 6 weeks after the infusion.All animals underwent liver biopsies before and 6 hours,1-6 weeks after operation.And contrast-enhanced abdominal CT was performed before and 6 weeks after operation.All animals were dissected 6 weeks after operation,aud each leaf of liver specimens were performed histological examination.Results There was no statistically significant difference of the portal venous pressure and diameter before infusion and 6 weeks after infusion in the experiment group 1 and control group (all P＞0.05).In the experiment group 2,compared with pre infusion,the portal vein pressure and diameter were higher than those of immediately and 6 weeks after infusion (all P＜0.05).In both experiment group 1 and group 2,all pigs had developed into liver fibrosis,the METAVIR score of 2 pigs in group 1 and 5 pigs in group 2 respectively were up to grade 4.Conclusion Portal infusion of 80％ alcohol is more suitable for establishing a swine model of liver cirrhosis with portal hypertension.

Objective To investigate the feasibility of reducing bolus?tracking monitor frequency in coronary CT angiography (CTA). Methods This prospective study including 120 patients with suspected coronary artery disease (CAD). According to the examination registration order, the patients were divided into groups A, B and C (n=40 for each group). All patients underwent coronary CTA with bolus?tracking technology, and were monitored at 10 s after the injection of contrast. The monitoring frequency of bolus?tracking for Group A was every 1.14 s, that for Group B was every 1.47 s , and for Group C was every 2.00 s, while the trigger threshold was set as 100 HU. To evaluate the image quality, the objective evaluation included signal noise ratio (SNR) and contrast noise ratio (CNR) of aorta (AO), CNR of left main coronary artery (LM) and right coronary artery (RCA), and the subjective score was recorded for each coronary artery segment. The monitoring times when CT density of the region of interest (ROI) reached the threshold, the CT value and the effective dose (ED) in the 3 groups were recorded. Objective image quality, monitoring parameters and radiation dose were compared using analysis of variance method, subjective image quality was compared withχ2 tests. Results There was no significant difference in AO (SNR and CNR), LM (CNR) and RCA (CNR) among the 3 groups, respectively (P>0.05). Subjective image quality scores of groups A, B, C were (1.879±0.042), (1.876±0.042), (1.881±0.042 ), with no significant difference (χ2=0.003,P>0.05). The monitoring times of to reach the threshold in groups A, B, C were (4.78±2.37), (3.76±1.39), (2.77±0.99), and ED were (0.058±0.031),(0.031±0.011), (0.021±0.007) mSv, with the significant difference (F=9.009, 31.998, respectively, P<0.01). Peak CT values during monitoring among three groups were (133 ± 24), (142 ± 39), (137±26) HU, respectively, with no significant difference (F=0.575,P=0.565). Conclusions It is feasible to reduce monitoring times when performing coronary CTA in dual?source CT scanner. The bolus?tracking monitor frequency in every 2 seconds can not only obtain satisfactory image quality, but also significantly reduce radiation dose.

Objective: To study the relationship between dilated cardiomyopathy and nuclear lamina protein (LMNA) gene mutation in Kazak ethnics at Xinjiang area.
Methods: A Kazak familial dilated cardiomyopathy (FDCM) with 31 members was studied. In addition, 160 patients with idiopathic dilated cardiomyopathy (IDCM) with 160 healthy controls were enrolled in our study, and they were divided into 4 groups: IDCM-Kazak, IDCM-Han and Control-Kazak, Control-Han.n=80 in each group. Peripheral blood DNA were extracted, 12 exons with nearby introns of LMNA gene were detected by PCR and the ampliifed products were sequenced and compared with the standard template of CHROMAS and BLAST software to identify mutation sites. LMNA mutation in both Kazak and Han IDCM patients were investigated.
Results: A novel LMNA mutation (insC, CGG→CCG) at exon 7 was identiifed in a FDCM proband, it caused an amino acid substitution as Arg to Pro, and a known LMNA polymorphism loci rs4641 (c.1362C>T His454His) was fund at exon 10. In addition, LMNA polymorphism loci rs4641 genotype distribution (χ2=5.16,P=0.036) and allele frequency (χ2=4.50,P=0.034) were statistically different between IDCM-Kazak group and Control-Kazak group; while such differences were no statistic meaning between IDCM-Han group and Control-Han group. Logistic regression analysis indicated that LMNA polymorphism loci rs4641 was related to IDCM occurrence in Kazak ethnics (P=0.025, OR=0.412, 95% CI 0.189-0.896).
Conclusion: LMNA polymorphism loci rs4641 was related to IDCM in Kazak ethnics at Xinjiang area, which might be susceptible loci for IDCM occurrence.