Objective To develop a superparamagnetic iron oxide (SPIO)-based MR probe containing vascular endothelial growth factor(VEGF) to investigate their biological and chemical properties and targeting effect of colorectal cancer cells in vitro. Methods The anti-VEGF-SPIO probe was fabricated with VEGF antibody and SPIO through chemical method. Its biological and chemical properties and reflexivity were tested with SDS-PAGE and MRL The SW620 cells incubated with anti-VEGF-SPIO probe for 30, 60 and 90 minutes respectively and compared with marrow mesenchymal stem cell at 37℃. The comparison among groups was conducted by using analysis of variance and LSD-t test. The MRI results were confirmed by the Prussian blue staining. The comparison among groups was performed by analysis of variance and factorial experiment. Results SPIO-based MR probe containing VEGF was successfully contributed and isolated. The reflexivity of anti-VEGF-SPIO probe was 0.0426×10~6 mol/s. The immunofluorescence and prussia blue stain proved high expression of VEGF in SW620 cells. Anti-VEGF-SPIO probe and SW620 cellscombined at 37℃ in vitro MRI proved the SW620 cells incubated with anti-VEGF-SPIO probe appeared hypointense on T_2WI and T_2~* WI. MR signal were 392±7,91±8,264±10 for 30, 60 and 90 minutes respectively, which were statistically different from that before incubation 679±12 (F=4735.489, P< 0.01). The intensity decreased most significantly at 60 minutes in vitro. Its MR signal 82±7 were statistically different compared with marrow mesenchymal stem cell 689±43, t=39.167,P<0.05). While SW620 cells incubated without SPIO were not statistically different compared with marrow mesenchymal stem cell, which were 419±59 and 400±41 respectively(t=-0.718,P>0.05). Conclusion Nanoscale iron particles containing the anti-vascular endothelial growth factor molecular probe can evaluate tumor angiogenesis at the receptor level, which provides a new way of the tumor angiogenesis diagnosis and anti-angiogenesis therapy.

Objective To evaluate the biochemical function of rat nature three-dimensional acellular pancreatic bioscaffold (3D-APB) in promoting cell proliferation and differentiation in vitro.Methods The fresh pancreas from 10 rats were perfused to prepare 3D-APB.The biocompatibility of 3D-APB was eva luated.The experiment was divided into 4 groups based on 4 kinds of three-dimensional media for AR42J culture,including blank control group,ECM group,PLGA group and 3D-APB group.We compared the proliferation and differentiation of AR42J pancreatic acinar cell at 3 days,5 days,7 days and 10 days after seeding among 4 groups.Results The 3D-APB could represent a biocompatible scaffold capable of integrating within host tissue.The proliferation rate of AR42J cells by MTT in 3D-APB was higher,while the apoptosis rate by flow cytometry was lower than those in other 3 media,which were all significantly different (P ＜ 0.05),respectively.The protein expression of PDX-1 and PTF-1 by western blot in 3D-APB group was greatly higher than those in other 3 groups (both P ＜ 0.05).The mRNA expression of PDX-1 and PTF-1 through qRT-PCR was significantly higher than those in other 3 groups (both P ＜ 0.05).Conclusion Compared with the commonly used chemical and natural scaffold at present,3D-APB could promote cell proliferation and differentiation,which was more appropriate for regenerative medicine.

Objective To investigate the distribution and drug resistance of pathogens for hospital-acquired urinary tract infections between patients in internal medicine wards and in surgical wards .Methods A total of 586 midstream urine samples were collected from patients in the First Municipal Hospital of Qinhuangdao during January 2012 and December 2014.Vitek 2 Compact system was applied in bacteria identification and drug sensitivity tests .Excel and SPSS 11.5 software were applied for data analysis . Results A total of 661 strains were isolated , in which 404 strains were from internal medicine wards and 257 strains were from surgical wards .Escherichia coli (44.6%vs.33.1%) and Enterococcus (23.0%vs. 16.3%) infections were more common in the internal medicine wards (χ2 =8.620 and 4.309, P<0.05), while the occurrence of Pseudomonas aeruginosa infection (4.0%vs.24.5%) was higher in surgical wards (χ2 =63.056, P <0.01).Escherichia coli and Klebsiella pneumonia strains were highly sensitive to piperacillin/tazobactam, cefptetan, amikacin, imipenem, and meropenem, and the sensitivity rates were from 85% to 100.0%.The sensitivity rates of Escherichia coli to ampicillin/sulbactam, levofloxacin and ciprofloxacin were <30%, and strains from surgical wards had lower sensitivity rates to these drug than those isolated from internal medicine wards (χ2 =4.987, 4.575 and 5.359, P<0.05).The sensitivity rates of Klebsiella pneumonia isolated from internal medicine wards to ceftazidime , gentamicin and aztreonam were 68.8%, 60.6% and 69.7%, which were higher than those isolated from surgical wards (36.0%, 32.0%, and 40.0%), and the differences were of statistical significance (χ2 =6.068,4.661 and 5.115, P<0.05).Pseudomonas aeruginosa strains were highly sensitive to piperacillin/tazobactam and amikacin, and the susceptibilities of strains isolated from surgical wards (98.4%and 96.8%) were higher than those isolated from internal medicine wards (75.0% and 81.3%) (χ2 =11.797 and 5.221, P <0.05). Pseudomonas aeruginosa strains isolated from surgical wards were also highly sensitive to cefepime (92.1%), but the sensitive rate of strains from internal medicine wards was only 37.5%, and the difference was of statistical significance (χ2 =24.696, P<0.01).Enterococcus faecium and Enterococcus faecalis were sensitive to tigecycline , vancomycin and linezolid with the sensitivity rates over 95%.Except quinupristin/dalfopristin and tetracycline , the sensitivities of Enterococcus faecalis to other antibiotics were higher than Enterococcus faecium.Susceptibility of Enterococcus faecium from surgical wards (33.3%) to moxifloxacin was lower than those from internal medicine wards (70.8%), and the difference was of statistical significance (χ2 =4.629, P <0.05).Conclusion There are differences in distribution and antimicrobial susceptibility of pathogens isolated from internal medicine wards and from surgical wards .

Aim To further research into the antithrombotic mechanism of scorpion venom active peptides (SVAP). Methods Human umbilical vein endothelial cells (HUVEC) were cultured with enzyme digestive method. After the cultured HUVC was incubated in conditioned media for 1 hour, the effects of SVAP on the concentration of 6-Keto-PGF 1? and NO of HUVEC were determined with radioactive-immunolygic and nitrate reduction enzyme method respectively. Results As compared with control, SVAP in the doses of 1,5,10, 20 mg?L -1 had the distinctive increase of 54%, 68%,72%,79% of the concentration of 6-Keto-PGF 1? and SVAP in the doses of 10, 20 mg?L -1 had the significantly increased of 27%, 46% of the concentration of NO. Regression anylysis showed that the release levels of PGI 2 and NO in HUVEC induced by SVAP was of positive correlation. Conclusion Antithrombotic mechanism of SVAP is related to the increase of PGI 2 and NO released from HUVEC and synergistic and mediating action between NO and PGI 2.

OBJECTIVETo observe the neurological protective effects of progesterone (PROG) on focal cerebral ischemia/reperfusion injury in rats and to explore its possible mechanism.

METHODSOne handred and twenty male SD rats were divided into three groups randomly: sham-operated group, middle cerebral artery occlusion ( MCAO ) group and PROG + MCAO group( n = 40). The right temporary MCAO model was established by the line-embolism method. The PROG + MCAO group rats were according to 8 mg/kg intraperitoneal injection PROG, after that 30 min, the rats were suffered ischemia/reperfusion. After rats were suffered ischemia for 2 h and reperfusion 0, 24, 48, 72 h stress, the nervous functional defect degree were evaluated by longe scoring, and the expression of two-pore domain K channel 3 (TASK3) mRNA in brain tissue were detected by the real-time PCR.

RESULTSPROG (8 mg/kg) could significantly reduced the nervous functional defect degree in rats after ischemia/reperfusion 24, 48, 72 h (P < 0.05). The results of real-time PCR showed that the TASK3 mRNA expression in the brain tissue at all time points significantly decreased in MCAO group compared with sham-operated group (P < 0.05). However, compared with MCAO group, the expression of TASK3 mRNA in brain tissue at all time points dramatically increased in PROG + MCAO group (P < 0.05).

CONCLUSIONPROG can improve the nervous functional defect degree after focal cerebral ischemia/reperfusion injury in rats, and the mechanism might be associated with up-regulating the expression of TASK3 mRNA in brain tissue.

Animals ; Brain ; drug effects ; pathology ; Brain Ischemia ; drug therapy ; Infarction, Middle Cerebral Artery ; Male ; Neuroprotective Agents ; pharmacology ; Potassium Channels, Tandem Pore Domain ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Reperfusion Injury ; drug therapy

There have been very few studies on the effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction in inhibiting apoptosis in myocardial ischemial injury caused by coronary heart disease. In this experiment, Gualou Xiebai Banxia decoction combined with-Xuefu Zhuyu decoction were used to intervene the miniature swine phlegm and blood stasis type coronary heart disease model, in order to observe the effect of the combined prescription on the myocardial apoptosis and the expressions of Bcl-2, Bax, Caspase-3, Caspase-9 in the model. Totally 15 Chinese experimental miniature swine were adopted and randomly divided into the control group, the model group and the phlegm and stasis-treating group. The model group and the stasis-treating group were fed with high fat diets for two weeks, intervened with the coronary artery injury and then given drugs and high fat diets for eight weeks. The control group was fed with ordinary diets for 10 weeks, without the coronary artery injury. After the experiment, myocardia at the juncture of infracted areas were collected and made into formalin-fixed paraffin sections. The TDT-mediate dUTP nick end labeling (TUNEL) assay was used to detect the myocardial apoptosis. The immunohistochemistry (IHC) technique was applied to detect Bcl-2, Bax, Caspase-3, Caspase-9 levels in myocardial tissues. According to the findings, the apoptosis indexes (AI) for the control group, the model group and the phlegm and stasis-treating group were 0.92%, 27.68%, 17.28%, respectively. The AI of the phlegm and stasis-treating group was significantly lower than that of the model group (P < 0.01). Compared with the model group, the phlegm and stasis-treating group showed significantly higher Bcl-2 protein expression (P < 0.01) and lower Bax, Caspase-3 and Caspase-9 protein expressions (P < 0.01). In conclusion, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction have a significant protective effect against the myocardial apoptosis in miniature swine phlegm and blood stasis type coronary heart disease model.
Animals ; Apoptosis ; drug effects ; Caspases ; metabolism ; Coronary Disease ; drug therapy ; Disease Models, Animal ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Male ; Medicine, Chinese Traditional ; Myocardium ; pathology ; Phytotherapy ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Swine ; Swine, Miniature ; bcl-2-Associated X Protein ; analysis

OBJECTIVEThe aim of the present study was to prepare uniform-sized silybin loaded poly (lactic-co-glycolic acid) (PLGA) microspheres in study of silybin with stainless steel membrane.

METHODSilybin PLGA microspheres were prepared by stainless steel membrane emulsification. The preparation conditions were optimized by single-factor test and orthogonal experiment, and evaluating the mean diameters, the particle size distribution, drug loading, entrapment efficiency and morphology of microsphere.

RESULTPrepared microspheres were round and surface was smooth. The mean diameter was (4.961 +/- 0.56) microm. The span was (1.75 +/- 0.18). The entrapment efficiency was (54.997 +/- 4.05)% and the average drug loading was (23.6 +/- 1.70)%.

CONCLUSIONThe stainless steel membrane emulsification can be used to prepare the silybin PLGA microspheres. The mean diameters of the silybin PLGA microspheres can be controlled in certain level. Stainless steel membrane emulsification has great potentiality exploitation and utilization.

Drug Compounding ; methods ; Emulsions ; chemistry ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry ; Silymarin ; chemistry ; Stainless Steel ; chemistry

Child ; Contracture ; congenital ; therapy ; Humans ; Male ; Neck Muscles ; pathology ; Torticollis ; congenital

OBJECTIVETo investigate effects of the variation of immune function in high humidity environment in different time, and lay a foundation for further study of the related mechanism.

METHODThirty SD rats were divided into 3 groups (n = 10): 20 day group, 40 day group in 90% relative humidity chamber and control group in normal relative humidity. Peripheral blood and spleens were collected to detect the levels of T lymphocyte subsets by Flow Cytometery.

RESULTSIn peripheral blood of the 20 day group rats, the CD3+ %, CD4+ %, CD8+ % and CD4+/CD8+ were 52.91 +/- 6.27, 37.80 +/- 4.11, 14.85 +/- 3.73 and 2.72 +/- 0.82 separately. Expect CD3+ %, they all had significant differences (P < 0.05). In addition, the data of the 40 day group rats showed no diversity in statistics. In spleen, CD8+ % of the 20 day group rats was 6.23 +/- 2.87 with significant differences (P < 0.05) and IgG, IgA and IgM did not change a lot in blood serum of the high humidity groups except C3 of the 20 days group (P < 0.05).

CONCLUSIONIn high humidity environment, the immune function of the rats increased in the initial stage. As time went on, the immune function gradually went to normal level through the self adjustment.

Acclimatization ; Animals ; Humidity ; Rats ; Rats, Sprague-Dawley ; Spleen ; immunology ; T-Lymphocyte Subsets ; immunology

Objective To explore the clinical results and complications of the traditional skeletal traction through olecranon on treating the irreducible humeral supracondylar fracture in children with of ulna.Methods Ninety-eight children patients of humeral supracondylar fracture with failure of reduction manipulation were selected as our subjects.The towel clamp-skeletal traction through olecranon of ulna was applied as the additional treatment methods.Meanwhile 5-24 months' follow-up were performed.Results According to Flynn elbow joint function evaluation standard.The curative effect was as followed.71 patients (74.7％) got the excellent outcome,17 patients (17.9％) for good outcome and 7 patients for improved utcome(7.3％).Therefore,the ratio of excellent operation reached to 92.6％.Conclusion The towel clampskeletal traction through olecranon of ulna might be an effective method to treat reliable fixation regarding of its high curing rate and simple process of performance.