Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.

Objective To evaluate the early appearance of SARS on chest CT scan and its role in the early diagnosis Methods Forty cases of SARS in keeping with the criteria of the Ministry of Health had chest CT scans within 7 days of onset of symptoms, and CR chest X-ray films were available as well These chest X-rays and CT images were retrospectively reviewed to determine if there were any abnormalities on the images The lesions on the chest CT images were then further analyzed in terms of the number, location, size, and density Results Positive abnormalities on chest CT scans were revealed in all 40 SARS cases Positive findings on CR chest films were showed in only 25 cases, equivocal in 6, and normal in 9 cases The main abnormalities seen on CT and X-rays were pulmonary infiltrations varied markedly in severity 70% cases had 1 or 2 lesions on chest CT scan, 30% cases had 3 or more lesions The lesions seen on chest CT scan tended to be ground-glass opacification, sometimes with consolidation which was very faint and inhomogeneous, easily missed on chest X-rays Typically the lesions were located in the periphery of the lung, or both central and peripheral lung, but very rare in a pure central location They were commonly in the shape of patch or ball Conclusions Chest CT scan is much more sensitive in detecting the lesions of the lung in SARS The early appearance of SARS on chest CT scan is characteristic but non-specific, indicating that chest CT scan plays a very important role in the early diagnosis and differential diagnosis of SARS

Objective To make a preliminary investigation and summary of the technique and efficacy of the novel intracranial stent, Enterprise, together with hydro-detachable coils for the treatment of very small intracranial wide-necked aneurysms (diameter<3 nun and body-to-neck ratio<1.5). Methods Six cases with very small intracranial wide-necked aneurysms were treated with Enterprise stents and hydro-detachable coils. In 5 cases the Enterprise stent was implanted to cover the neck of the aneurysm, which was followed by the introduction of a microcatheter into the aneurysmal sac through the stent mesh to stuff hydro-detachable coils in order to fill the aneurysmal sac. In the remaining case, the microcatheter was placed into the aneurysmal sac before the Enterprise stent was inserted to embolize the aneurysm. Postoperative follow-up was conducted for 3-6 months. Results The operation was successfully completed in all 6 patients, with the implanted stents being in right place. The parent arteries remained patency in all patients. No complications occurred. Complete occlusion of aneurysmal cavity was obtained in four cases, and the occlusion degree of the aneurysmal cavity above 95% was seen in 2 cases. After the procedure, all the patients recovered well. Neither rebleeding nor symptoms related to thrombosis occurred during a clinic follow-up of 3-6 months. Conclusion Endovasculur embolization with Enterprise stent together with hydro-detachable coils is a safe and effective method for the treatment of very small intracranial wide-necked aneurysms. However, its long-term effect needs to be further observed.

Objective To investigate the effect of Amp activated protein kinase(AMPK)activity in radiosensitivity of human na-sopharyngeal carcinoma cells .Methods Human nasopharyngeal carcinoma cells ,CNE-2 ,were treated with AMPK inhibitor ,Com-pound C(CC) ,for 1 h .Then cells were explored in X-ray .The expression of total AMPK (t-AMPK) ,phosphorylation AMPK (p-AMPK) ,and MAPlLC3 were detected by Western blot .The number of autophagosomes were observed and calculated by transmis-sion electron microscope(TEM) .Cells processed with CC were explored in X-ray .MTT assay was used to detect the difference of two groups in cell proliferation .Cell apoptosis were assayed by flow cytometry .Results The expression of p-AMPK in CC group cells were significantly downregulated compared to the negative control group cells (P< 0 .01) ,while no significant change of t-AMPK expression were found(P>0 .05) .The expression of MAPlLC3 and the number of autophagosomes in CC group cells were significantly decreased compared to the control group cells (P<0 .05) .Correspondingly ,the cell proliferation rate in CC group was lower than in control group ,and the percent of apoptosis cells was higher in CC group than in control group (P<0 .05) .Conclusion Suppression of AM PK activity could inhibited autophagy induced by decreasing the degree of p-AM PK ,then enhanced the effect of proliferation inhibition and apoptosis promotion in CNE-2 cells .The AMPK inhibitor ,CC ,can serve as an effective assistant treatment of radiotherapy for nasopharyngeal carcinoma .

Objective To analyze and compare images of radial head fracture of 50 patients acquired by computed radiology(CR),coronal plane and axial plane of CT scan.And to determine routine plane of CT scan for radial head fracture.Methods Images of of radial head were acquired by CR,coronal plane and axial plane of CT scan on 50 patients with radial head fracture initially diagnosed by orthopedists. classify all the cases of radial head fracture into type Ⅰ、Ⅱ、Ⅲ and Ⅳ according to the classification proposed by Mason.Results The positive incidence of CT and CR were 96%(48)and 78%(39) respectively.Cases of 94%o(47)through CT coronal scan and 82%(41)eases through CT axial scan were exactly classified.Conclusion The designation of the plane of CT scan is significant to the classification of the radial head fracture.Coronal plane CT scan can meet the need of imaging clinical classification and is recommended to be routine plane of radial head fracture.In order to ensure the exact classification axial plane and 3D reconstruction technique should be added for type Ⅲ and type Ⅳ of radial head fracture.

Objective To explore the inhibitory effect of melatonin(MT) on expression of glial fibrillary acidic protein in infant rats with brain injury induced by lipopolysaccharide(LPS).Methods One hundred and sixty Spragne-Dawley rats(1 month) were randomly divided into 4 groups:9 g/L saline group(NS group),LPS group,MT protective A group(MT plus LPS,MT was given 15 minutes earlier),and MT protective B group(MT plus LPS,MT was given immediately).Forty rats in each group,then every group was subdivided into 2 subgroups according to observe time point at 24 h and 48 h with 20 rats,respectively.Half of them in all subgroups were not injected Evans blue(EB).Rats were sacrificed at 2 time points to take brain tissue samples,respectively.EB content of brain tissue was meteraged by the formamide methods.Immunohistochemistry technology and computer assisted image analysis system methods were applied to observe the expression of the positive apoptosis cell of brain tissue.Results Both EB content and the expression of GFAP showed significant difference between LPS group and NS group(Pa0.05).Conclusions LPS can induce the expression of GFAP in infectious brain injury of rats induced by LPS,and MT can inhibit the expression of GFAP;MT may play a role in dealing with infectious brain injury.

Three-dimensional (3D) culture of cells could closely mimic the in vivo situation with regard to cell function and microenvironment compared with plane monolayer cultured cells. In this paper, we established 3D culture of rat WB-F344 cells with rotary cell culture system (RCCS) to simulate microgravity environment, and examined cells proliferation, morphology, microstructure, E-cadherin protein quantity and mRNA expression of adhesion molecules by count the number of cells, optical microscope, transmission electron microscope and reverse transcriptase-polymerase chain reaction (RT-PCR). The results demonstrated that cells were polyhedron with lots of micovilli and mitochondria, which grow well and packed together densely to form irregular aggregates. Adjacent cells were connected with desmosome and tight junction. With the regard, the aggregates behaved 3D growth characteristics. Moreover, compared with control, mRNA level of Fibronectin and E-cadherin protein were increased, the changes maybe is the part mechanism in this microgravity simulated cells culture models which strengthened cells junction. This rotating 3D model might facilitate the study of interactions of cell-cell, cell-matrix and the mechanisms.
Animals ; Cadherins ; genetics ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cell Proliferation ; Fibronectins ; genetics ; Rats ; Spheroids, Cellular ; ultrastructure ; Weightlessness Simulation

OBJECTIVETo compare the effects and mechanism of blood enriching on mouse model of blood deficiency syndrome induced by cyclophosphamide of albiflorin and paeoniflorin.

METHODAlbiflorin and paeoniflorin were determined by using animal models of blood deficiency syndrome induced by cyclophosphamide. The amount of WBC, RBC, HGB, index of thymus gland and spleen, and the changes of GM-CSF, IL-3 and TNF-α in serum were detected after the treatment.

RESULTCompared with the model group, the amount of WBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01). The amount of RBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01, P < 0.001), which did not had a significant difference compared with the same dose. The index of thymus gland in the group of 30 mg x kg(-1) albiflorin was superior to the model group (P < 0.01), the difference was significant compared with the same dose of paeoniflorin (P < 0.05). The GM-CSF in serum in all groups of 30 mg x kg(-1) albiflorin, 15 mg x kg(-1) albiflorin, 30 mg x kg(-1) paeoniflorin and 15 mg x kg(-1) paeoniflorin increased obviously (P < 0.01, P < 0.05, P < 0.01, P < 0.05); The IL-3 in serum in both group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin also increased (P < 0.001). The content of TNF-α in group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were reduced (P < 0.01), which showed the obvious difference compared with the same dose group (P < 0.01).

CONCLUSIONAlbiflorin had the effect of blood enriching by regulating the immune function, same with the paeoniflorin. The probable mechanism of nourishing blood and liver of Paeoniae Radix Alba was not only the better effect of adjusting the content of TNF-α, but also might act synergistically with paeoniflorin.

Animals ; Blood Cells ; drug effects ; Bridged-Ring Compounds ; pharmacology ; Cyclophosphamide ; toxicity ; Glucosides ; pharmacology ; Granulocyte-Macrophage Colony-Stimulating Factor ; blood ; Hematopoiesis ; drug effects ; Interleukin-3 ; blood ; Male ; Mice ; Monoterpenes ; pharmacology ; Tumor Necrosis Factor-alpha ; blood

Objective To understand detection results and difference in multidrug-resistant organisms(MDROs) in intensive care unit(ICU)and non-ICU.Methods Strains isolated from clinical specimens of hospitalized patients in a hospital from January 2015 to December 2016 were analyzed, 6 kinds of MDROs were conducted targeted monitoring, isolation and antimicrobial resistance of 6 kinds of MDROs from ICU and non-ICU patients were compared. Results A total of 1 013 strains of 6 kinds of MDROs were monitored, isolation rate was13.13％.Isolation rate of MDROs in ICU was higher than that of non-ICU (24.60％vs 5.47％, P＜0.001).Carbapenem-resistant Acinetobacter baumannii(CRAB)was the main isolated MDROs, accounting for 69.40％;of different pathogenic organisms, isolation rate of CRAB was the highest(55.75％).The main MDROs detected in ICU and non-ICU were both CRAB, accounting for 76.32％and 48.62％respectively;Of isolated pathogens, isolation rate of MDROs in ICU was higher than that of non-ICU(47.95％vs 8.02％, P＜0.001).Antimicrobial resistance rates of Escherichia coli isolated from ICU to ticarcillin/clavulanic acid, ceftriaxone, cefotaxime, cefepime, imipenem, meropenem, amikacin, and gentamicin were all higher than that of non-ICU, resistant to piperacillin was lower than non-ICU, difference was statistically significant(all P≤0.05);resistance rates of Klebsiella pneumoniae from ICU to common antimicrobial agents(except piperacillin)were all higher than non-ICU(all P＜0.05).Resistance rates of Acinetobacter baumannii and Pseudomonas aeruginosa from ICU to common antimicrobial agents were all higher than non-ICU (all P＜0.05).Resistance rates of Staphylococcus aureus isolated from ICU to oxacillin, ciprofloxacin, tetracycline, and rifampicin were all higher than non-ICU (all P＜0.05), and resistance rates of Enterococcus faeciumto quinupristin/dafoeleptin and tetracycline were both lower than non-ICU (both P＜0.05).Conclusion Isolation rate of MDROs in ICU is high, resistance rates to most antimicrobial agents are also higher than non-ICU, monitoring on MDROs in ICU should be strengthened, and according prevention and control measures should be formulated.

Objective To explore the effects of epithelial cell adhesion molecule(EPCM) expression downregulation on the proliferation,invasion and drug sensitivity of ECAM+/cluster of differentiation 44 (CD44) + colorectal cancer stem cells.Methods Colorectal cancer stem cells (ECAM) were obtained from human colorectal cancer cell line LoVo by the method of tumor microspheres.The specific markers of colorectal cancer stem cells were identified and studied.Lipofectamine 2000 liposome was used to complete the transfection,which was divided into three groups:blank group (ECAMhjgh/CD44 + LoVo),control group (transfection of common inhibitors,lipofectamine 2000-inhibitors) and experimental group (ECAM) according to different treatment methods.They were cultured in the same way,the three group of cells in logarithmic phase were treated with irinotecan (drug concentrations were 1,5,10,15,20,25,30,35,40,45,50 mg · L-1) and capecitabine (drug concentrations were 50,100,150,200,250,300,350,400,450,500,550,600mg· L-1).Then,they continued to be cultured.The expression levels of ECAM mRNA in three groups of ceils were detected by real-time fluorescence quantitative detection (RT-PCR).The thiazolyl blue tetrazolium bromide (MTI)assay was used to detect the proliferation and drug sensitivity of the three groups before and after treatment with irinotecan and capecitabine.Transwell cells were used to detect the invasion ability of three groups of cells.Results The percentage of ECAMhjgh/CD44 + colorectal cancer stem cell in LoVo cell line was 0.95％,85.78％ before and after enrichment,and the difference was significant (P ＜ 0.05).The expression of ECAM mRNA in blank group,control group and experimental group were 8.17 ±0.64,7.94 ±0.83,2.16 ±0.12.Compared with blank group,the difference had significantly in two groups (P ＜ 0.05,P ＜ 0.01).Compared with control group,the difference in experimental group had significantly in two groups (P ＜ 0.05).It indicated that model was succeed prepared.Invasive cell in the three groups were 79.22 ± 5.25,80.12 ± 4.89,31.23 ± 2.36.Compared with blank group,the difference had significantly in two groups (P ＜0.05,P ＜0.01).Compared with control group,the difference in experimental group had significantly(P ＜0.05).The IC50of irinotecan on colorectal cancer stem cells in the three groups were (20.25 ±4.35),(19.22 ±3.99),(10.24 ± 2.04) mg · L-1.The IC50of capecitabine on colorectal cancer stem cells in the three groups were (320.13 ± 23.65),(315.79 ± 21.03),(250.22 ± 15.45) mg · L-1.Compared with control group,the difference in experimental group had significantly(P ＜0.05).Conclusion Targeted down-regulation of ECAM can effectively inhibit the proliferation and invasion of colon cancer stem cells,and enhance their sensitivity to drugs at the same time.