To construct expression vectors of small hairpin RNA aimed at N-acetylglucosaminyltransferase Ⅴ(GnT-Ⅴ) gene, and to investigate effects of GnT-Ⅴ shRNA on proliferation, adhesion, migration and invasion of LoVo cell line. siRNAs were designed according to the coding sequence of GnT-Ⅴ gene, shRNA expression vectors were constructed and transfected into LoVo cell line, cell lines which stably expressed low level of GnT-Ⅴ were established by G418 screening. The mRNA and protein expression of GnT-Ⅴ were measured by semi- quantitative reverse transcription polymerase chain reaction(RT-PCR) and Western blot analysis, respectively. The effects of pGPU6/GFP/Neo GnT-Ⅴ shRNA vectors on proliferation, adhesion, migration and invasion of LoVo cell line were evaluated by CCK-8 assay, heterogenous adhesion, wound closure assay, chemotactic migration and cell invasive experiment, respectively. GnT- Ⅴ shRNA expression plasmid was constructed successfully and pGPU6/GFP/Neo GnT-Ⅴ shRNA down-regulated expression of GnT-Ⅴ dramatically in LoVo cell. Expression of LoVo GnT-Ⅴ/1564 and LoVo GnT-Ⅴ/2224 dereased by 82%, 71.5% respectively at mRNA level, and 68%, 56% respectively at protein level. The more effective interfered cell line, LoVo GnT-Ⅴ/1564, was chosen to do further experiment. CCK-8 assay showed proliferation of LoVo GnT- Ⅴ/1564 was suppressed obviously, compared to proliferation of negative control group cell (P

Objective To compare the clinical outcomes of urethral resectoscopy and retroperitoneal laparoscopic nephroureterectomy with open surgical nephroureterectomy. Methods A total of 44 patients with renal pelvic and ureteral neoplasms were included.Urethral resectoscopy and retroperitoneal laparoscopic nephroureterectomy were performed in 15 cases (Group A) and open surgical nephroureterectomy in 29 cases (Group B).The therapeutic effectiveness,postoperative recovery,relevant cost and complications were compared between Group A and Group B. Results The analysis showed that the intra-operative bleeding volum [(75.1?29.5)ml],the postoperative intesinal function recovery [(24.1?12.6)h],time to ambulation [(24.3?10.5)h],use of antalgesic [(3.0?0.8)d],intravenous antibiotic [(7.2?3.1)d],hospital stay [(6.3?1.2)d],convalescene of normal activities [(28.0?7.8)d] and incidence of complications with Group A were significantly superior to those with Group B(P

Objective:To introduce the fabrication of lead eyeshade and its use in the radiotherapy of both malignant and benign eye tumors and observe the preliminary clinical effect. Methods:To lead sheet with thickness of 2.5-3mm was fabricated into spherical eyeshade and erythromycin Eye Ointment was smeared onto it. Results:According to EORTC criteria, the first level side effect was occasional and mild pain and drying of the eye. The second level was intermittent and tolerable pain and drying of the eye. The third level was constant and intense pain and drying of the eye and the fourth level was incurable and intolerable and drying of the eye. Conclusion:It was shown that the use of lead eyeshade can not only ensure the efficacy of radiotherapy, but also reduce the incidence of radiation injury of surrounding normal tissues. The method used for making of lead eyeshade is effective and easy to grasp.

Objective To investigate the effects of Lp(a)on proliferation GMCs of rat model induced by lipopolysaccharide and explore the possible mechanism of Lp(a)in the proliferation of rat GMCs.Methods To observe the effects of Lp(a)on proliferation of GMCs,different dosage of Lp(a)were used,The research were divided into three groups:Control group,LPS group,Lp(a)group.After culture(at the end of 12h,24h,48h,60h and 72h),the cultured GMCs and suspension were collected to observe the rate of GMCs proliferation by MTT,the positive rate of proliferation cell nuclear antigen(PCNA)by immunohistochemisty,and the level of intercellular adhesion molecule-1(ICAM-1)by ELISA respectively.Results Compared with control and LPS group,MTT,positive rate of PCNA and ICAM-1 of GMCs were increased more significantly in Lp(a)group.MTT ,the positive rate of PCNA and ICAM-1 of GMCs were increased as Lp(a)dosage increased,a maximal effect was seen when Lp(a)was 2.5 μg/L or 5.0μg/L.When the dosage continue increased,MTT,the positive rate of PCNA and ICAM-1 activity of GMCs began to decrease in Lp(a)group.ICAM-1 showed positive correlation with MTT and the positive rate of PCNA.Conclusion Lp(a)can significantly affect the rate of GMCs proliferation,and this affection is in a dosage-and timedependent manner.Low dosage stimulates GMCs proliferation, and high dosage inhibits GMCs proliferation.ICAM-1 shows positive correlation with MTT and the positive rate of PCNA.The effect of Lp(a)on GMCs may be through ICAM-1.

This study was aimed to work out a simple, applicable, sensitive and specific protocol for the identification of phosphoproteome. Isotope-labeling, two-dimensional electrophoresis, autoradiography and so on were used to establish a phosphoproteome map of mice neurons, and then chemiluminescence Western blotting was utilized to detect three phosphoproteins PI3Kr3, MEK1 and PKCalpha selectively. The results of comparison showed that the blots of PI3Kr3, MEK1 and PKCalpha on autoradiography map were almost identical with the blots of PI3Kr3, MEK1 and PKCalpha on chemiluminescence Western blotting maps. So this protocol based on isotope labeling and chemiluminescence Western blotting methods has proven to be sensitive and specific in the identification of phosphoproteome.
Animals ; Animals, Newborn ; Blotting, Western ; methods ; Brain ; cytology ; Cells, Cultured ; Electrophoresis, Gel, Two-Dimensional ; methods ; Luminescent Measurements ; methods ; Mice ; Neurons ; chemistry ; cytology ; Phosphoproteins ; analysis ; Phosphorus Radioisotopes ; Proteome ; analysis ; Sensitivity and Specificity

Objective:To investigate the correlations of human leukocyte antigen (HLA)-A, B, C, and DRB1 genotypes with cross-reactivity caused by aromatic antiepileptic-drugs.Methods:A case-control association study was carried out on subjects who accepted treatments/physical examination in our hospitals from September 2016 and September 2020; 31 patients with aromatic antiepileptic drugs (carbamazepine, phenytoin, oxcarbazepine, lamotrigine and phenobarbital)-induced cross-reactivity were enrolled as patient group, 52 tolerant subjects who took the 5 antiepileptic drugs for more than 3 months without cross-reactivity were chosen as tolerant control group, and 500 healthy volunteers were recruited as normal control group. The ethnicity of all patients and controls was Han Chinese. High-resolution genotyping was performed to compare the HLA-A, B, C, and DRB1 genotypes in subjects of the 3 groups. χ2 test or Fisher's exact test were used to analyze the correlations of HLA genes with cross-reactivity caused by aromatic antiepileptic-drugs. Results:The presence of HLA-B*13:01 genotype in the patient group, the tolerant control group, and the normal control group was 45.2% (14/31), 15.4% (8/52) and 14.6% (73/500), respectively. The presence of HLA-B*13:01 genotype in the patient group was significantly higher as compared with that in the tolerant control group and normal control group ( Pc<0.017). No other HLA genotypes were found to be associated with cross-reactivity caused by aromatic antiepileptic-drugs. Conclusion:HLA-B*13:01 is the risk genotype for cross-reactivity caused by aromatic antiepileptic-drugs.

Objective To establish a stable and feasible rabbit model of cardiopulmonary bypass (CPB) in acute cerebral embolism phase for studying the effects of CPB on brain tissues and the timing of surgical intervention of acute cerebral embolism. Methods Fifty-four rabbits were randomized into group A (n=18) to receive CPB without middle cerebral artery occlusion (MCAO) and group B to undergo CPB at 24 h (group B1, n=18) or 1 week (group B2, n=18) after MCAO. Through a supraorbital margin approach, electrocoagulation was carried out to occlude the main stem of the left MCA under direct vision to establish MCAO. Magnetic resonance imaging (MRI) was performed at both 24 h and 1 week after MCAO, and the severity of cerebral embolization was evaluated. CPB was established by cannulation of the ascending aorta and the right atrium through a median sternotomy incision. MRI was performed at 2 h after CPB to observe the brain tissues. Results MCAO was successfully established in groups B1 and B2, and all the rabbits survived after MCAO. In both groups A and B, MRI examination detected no cerebral hemorrhage or new embolism 2 h after CPB. Conclusions We have established a stable and feasible CPB model in rabbits with acute cerebral embolism to allow study of the mechanisms of CPB-related organ damage and its interventions.

Objective To establish a stable and feasible rabbit model of cardiopulmonary bypass (CPB) in acute cerebral embolism phase for studying the effects of CPB on brain tissues and the timing of surgical intervention of acute cerebral embolism. Methods Fifty-four rabbits were randomized into group A (n=18) to receive CPB without middle cerebral artery occlusion (MCAO) and group B to undergo CPB at 24 h (group B1, n=18) or 1 week (group B2, n=18) after MCAO. Through a supraorbital margin approach, electrocoagulation was carried out to occlude the main stem of the left MCA under direct vision to establish MCAO. Magnetic resonance imaging (MRI) was performed at both 24 h and 1 week after MCAO, and the severity of cerebral embolization was evaluated. CPB was established by cannulation of the ascending aorta and the right atrium through a median sternotomy incision. MRI was performed at 2 h after CPB to observe the brain tissues. Results MCAO was successfully established in groups B1 and B2, and all the rabbits survived after MCAO. In both groups A and B, MRI examination detected no cerebral hemorrhage or new embolism 2 h after CPB. Conclusions We have established a stable and feasible CPB model in rabbits with acute cerebral embolism to allow study of the mechanisms of CPB-related organ damage and its interventions.

Objective:To assess the feasibility ,safety and short‐term outcome of simultaneous integrated boost‐intensity modulated radiation therapy(SIB‐IMRT) with concurrent capecitabine chemotherpay for anal cancer .Methods:A total of 10 hospitalized patients with anal cancer during Sep .2009 and Feb .2014 were treated with SIB‐IMRT .A total dosage of 57 .6 Gy was given to the primary lesion and macroscopical lymph nodes in 32 fractions ,with 1 .8 Gy in each fraction .And a total dosage of 48 Gy was given to the bilateral iliac vessels and inguinal lymphatic drainage region in 32 fractions ,with 1 .5 Gy in each fraction .And capecitabine was concurrently administered at the oral dose of 625 mg/m2 ,twice daily ,5 days per week . Two patients received a sequential radiation boost dose of 2 × 1 .8 Gy due to macroscopic residual lesion at week 5 of treatment . Acute and late adverse reaction was assessed according to the Common Terminology Criteria for Adverse Events version 4 .0 . Results:All patients completed radio‐chemotherapy without any treatment break .The incidence rate of grade 3 skin adverse reaction was 50% (5/10) .No grade 4 adverse reaction was observed .Mean follow‐up was 20 months(range 6‐60 months) .The 2‐year‐local control ,colostomy‐free survival ,distant metastases‐free survival and overall survival rates were 100% (10/10) ,90%(9/10) ,90% (9/10) ,and 90% (9/10) ,respectively .Conclusions:SIB‐IMRT with concurrent capecitabine chemotherapy :an acceptable safe regimen ,however ,more samples and a longer follow‐up are required to assess its potential superiority .

Objective:To explore the effects of perioperative comprehensive nursing program on the postoperative management of patients with low-grade glioma in sports area and epilepsy.Methods:Totally 130 patients with low-grade glioma in sports areas and epilepsy who chose to undergo craniotomy at the No.12 Ward Department of Neurosurgery in Beijing Tiantan Hospital, Capital Medical University from May 2018 to April 2019 were selected and divided into the observation group ( n=74) and the control group ( n=56) according to the admission time. Patients in the observation group adopted the comprehensive nursing program, while patients in the control group received routine care. Skin redness events, falls/falling out of bed, average length of hospital stay and patient satisfaction of the two groups of patients were observed and statistically analyzed. Results:The incidence of skin redness in the observation group was lower than that in the control group (4.05% vs. 16.07%, P=0.030) ; the incidence of falls/falling out of bed in the observation group was lower than that in the control group (2.70% vs. 12.50%, P=0.015) ; the average length of hospital stay in the observation group was lower than that in the control group [ (7.6±0.3) vs. (11.3±0.5) d, P< 0.01]; and the differences were all statistically significant. Conclusions:Comprehensive nursing can reduce the incidence of skin redness and falls/falling out of bed events in patients with low-grade glioma in sports areas and epilepsy, shorten their average postoperative hospital stay, and improve their satisfaction.