Objective To modify the methods of operation and establishment of cerebral ischemia rat models made by thread occlusion. Method We randomly divided 120 male SD rats into a common group (n = 50), an improvement group (n = 60) and a sham-operated group (n - 10). Rats in the common and improvement groups were made into models using the common and improvement methods separately. All models were evaluated on the basis of physical sign indices and 2,3,5-triphenyltetrazolium chloride (TTC) staining. The TTC staining results were taken as gold standards. Then, we compared the achievement ratios of the two groups, and computed the sensitivity and specificity of every physical sign index based on these standards. The χ~2 or correction χ~2 test was used to compare the ratios. Results (1) The achievement ratio in the improvement group was significantly higher than that in the common group (71.67% vs. 52.00%, P = 0.034). (2) The sensitivity of evaluation for both common and improvement methods was 98.55%. However, the specificity of evaluation for the improvement method was significantly higher than that for the common method (100.00% vs. 40.00%, P =0.000). Conclusions The establishment achievement ratio and evaluation correctness of models are obviously elevated by modification of the operation and establishment methods.

Objective To investigate the effect of treating tibial plateau fractures involving the posterior column via anterolateral approach.To discuss its merit and demerit,indications and contraindications.Methods From Jan 2012 to Jan 2015,37 patients with closed tibial plateau fractures involving the posterior column were treated by 3.5mm proximal tibial plate with narrow plate head through standard anterolateral approach.The group included 25 males and 12 females,aged from 23 to 65 years old (average 44.6 years old).During the treatment and follow-up period,the curative effect was evaluated by using Rasmussen's radiological grading for early radiological outcomes and HSS grading,Lysholm grading,Lachman test and Pivot-shift testfor clinical examination at the 1 year follow-up.Results The average time for the operation was 98 min (range,55-170 min).1 year follow-up was completed in 37 patients.The average fracture union time and full weight bearing were 10.6 weeks (range,8-16 weeks) and 11.2 weeks (range,8-20 weeks) respectively without reduction loss.No statistical difference was found in either the tibial plateau angle (TPA) or posterior slope angle (PA) when comparing the results at instant,3rd month,6th month and 12 month.The mean score of the Rasmussen' s radiological grading was 15.6(range,12-18) and the mean score of the HSS grading was 88.6 (range,80-100).The average range of motion of the knee joint was 128.6°(range,110°-150°).The mean score of the Lysholm grading was 91.6±3.9. The Lachman test and the Pivot-shift test were negtive.Conclusion Treating tibial plateau fractures involving the posterior column by 3.5 mm proximal tibial plate with narrow plate head through standard anterolateral approach is an effective method.The protocol is simple and safe,the approach is familiar by most clinicians.Good reduction and fixation,earlier functional exercise can be achieved easily.The knee function recovered well and earlier curative effect was satisfied.

Objective To evaluate the frequency and morphological characteristics of the posterolateral articular fracture in tibial plateau fractures.Methods A retrospective analysis of CT images and clinical data was taken among 309 cases of tibial plateau fractures from 2008 May to 2013 January.There are total 264 patients were recorded excluding 45 cases with which the CT image is missing or not compatible with the medical Picture Archiving and Communication Systems (PACS).To determine the occurrence rate of the posterolateral articular fracture in tibial plateau fractures and measure morphological parameters such as the axial angle of fracture line,articular surface area,sagittal fracture angle,fracture height,and amount of displacement.Results 39 cases of posterolateral articular fragments were found in 264 cases of tibial plateau fractures with the 14.8％ incidence (39/264).There were 18 males and 21 females,aged from 31-70 years (mean,52 years).17 left cases and 22 right cases.The mechanism of injury were traffic accident in 22 patients,blow by a heavy object in 2 patients,a fall in 11 patients,and other causes (unknown) in 4 patients.The posterolateral plateau articular fracture fragment has morphological characteristics of a conical shape fragment and the major articular fragment angle was 22° (range,-43°-62°),and an average accounted for 14.1％ of the articular surface of the total tibial plateau (range,8％-32％).The posterolateral fragment exhibits a vertical fracture pattern and an average sagittal fracture angle was 76° (range,58°-97°),suggestive of shear instability and vertical displacement.Maximum posterior cortical height was 28 mm (range,18-42 mm),and average size of the displacement was 10.48 mm (range,2-19 mm).Conclusion The posterolateral plateau articular fracture fragment has morphological characteristics of a conical shape fragment,relative small articular surface area and sagittal fracture angle.

Objective To evaluate the role of interleukin?1β( IL?1β) in activation of N?methyl?D?aspartate ( NMDA) receptors in spinal dorsal horns of rats with neuropathic pain. Methods One hundred twenty?eight pathogen?free healthy adult male Sprague?Dawley rats, weighing 200-250 g, were divided into 4 groups (n=32 each) using a random number table: control group (group C), recombinant adenovirus vector group ( group rAd) , recombinant adenovirus vector shRNA group ( group rAd∕shRNA) , and recom?binant adenovirus vector shRNA plus recombinant adenovirus vector IL?1β group ( group rAd∕shRNA+rAd∕IL?1β) , receiving intrathecally injected equal volume of 0.9% sodium chloride solution, recombinant ade?novirus vector, recombinant adenovirus vector shRNA, and recombinant adenovirus vector shRNA plus re?combinant adenovirus vector IL?1β5 μl, respectively. Chronic compression of dorsal root ganglia was per?formed on 8th day after intrathecal injection. The thermal paw withdrawal latency ( TWL) was measured at 1 day before intrathecal injection ( T0 ) and at 1, 2, 3 and 5 weeks after intrathecal injection ( T1-4 ) . After measurement of the pain threshold at each time point, 8 rats selected randomly were anesthetized and sacri?ficed, and the lumbar enlargement segments of the spinal cord were harvested for determination of the ex?pression of IL?1β and phosphorylated NMDA receptor NR1 subunits at serine 896 ( pNR1S896) in spinal dorsal horns on the injured side by immunohistochemistry and Western blot. Results Compared with group C, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA (P<0.05), and no significant change was found in the parameters mentioned above at each time point in group rAd and group rAd∕shRNA+rAd∕IL?1β( P>0.05) . Compared with group rAd, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA ( P<0.05) , and no significant change was found in the parameters mentioned above at each time point in group rAd∕shRNA+rAd∕IL?1β (P>0.05). Compared with group rAd∕shRNA, the TWL was significantly short?ened, and the expression of IL?1β and pNR1S896 in spinal dorsal horns was significantly up?regulated at T2-4 in group rAd∕shRNA+rAd∕IL?1β ( P<0.05) . Conclusion IL?1β is involved in the activation of NM?DA receptors in spinal dorsal horns of rats with reuropathic pain.

Objective To study the relationship between environmental chemical elements,vires infection and dilated cardiomyopathy(DCM).Methods In 2008,233 patients with DCM(case group)and 150 patient with stable angina(control group)were chosen in Liaocheng People's Hospital and Yanggu People's Hospital,Shandong province.Population distribution and disease history were surveyed in the two groups.Human myocardial antibody IgG(AMA-IgG),Coxsackie B virus IgG(CBV-IgG),Adenofirus antibody IgG(ADV-IgG)were detected by ELISA in both the case group and the control group.Serum trace elements were detected in the two groups.The general chemical and toxicological indicators in drinking water of the high-and the low-incidence aireas of the disease were control group[60.00%(90/150),χ2=13.80,P<0.01)].Per capita annual income(Yuan,RMB)in the case group (3207.82±618.51)was lower than that of the control group[(5086.61±886.12),t=24.40,P<0.01].Personal alcohol consumption in the case group[(175.00±160.50)g/d]was higher than that of the control group[(110.22±100.03)g/d,t=4.40,P<0.01)].The rate of myocarditis in the case group[5.15%(12/233)]was higher than ADV-IgG in the cage group were 7.78%(7/90),6.67%(6/90)and 6.67%(6/90),respectively.Compared with those in the control group[3.33%(2/60),5.00%(3/60)and 5.00%(3/60),χ2=1.26,0.18,0.18,all P>0.05],no mg/L]in drinking water of the high-incidence areas were significantly higher than that of iron[(0.39±0.67)mg/L,t=2.11,P<0.05]and that of manganese[(0.15±0.14)mg/L,t=3.01,P<0.01]in the low-incidence arefas.The content of semm iron[(69.1±57.8)μmol/L]in the case group evidently exceeded the normal range(15.6-35.9 μmol/L)and obviously higher than that in the control group[(20.0±17.5)μmol/L,t=5.04,P<0.01].Conclusions Theso data do not support that DCM is related with persistent virus infection and autoimmunization.DCM is probably related with low incomes,high alcohol consumption,myocarditis,high iron and manganese contents in drinking water and high content of serum iron.

OBJECTIVETo compare the durability of quantum dots with that of green fluorescein for labeling survivin antisense oligonucleotide (ASODN) and investigate the difference in growth and apoptosis of cells transfected with the labeled survivin ASODN.

METHODSSurvivin ASODN labeled with quantum dots or green fluorescein was transfected into MCF-7 cells via Lipolifectmain(TM2000). The proliferation of MCF-7 cells was assessed with MTT assay, survivin mRNA expression determined by RT-PCR and its protein expression measured by Western blot analysis. The apoptosis rate of the transfected cells was estimated by flow cytometry, and the fluorescence distribution in the cells observed under fluorescent inverted microscope.

RESULTSThe mRNA and protein expressions of survivin were significantly decreased in the MCF-7 cells after cell transfection with survivin ASODN labeled with quantum dots or green fluorescein, and no significant difference was noted between the two labeling methods (P>0.05). Nor did survivin ASODN transfection with different labeling methods produced significant difference in cell proliferation and apoptotic rate (P>0.05). For green fiuorescein labeling, the fluorescence disappeared 4 days after transfection, whereas the fluorescence sustained for 1 week for quantum dots labeling.

CONCLUSIONSurvivin ASODNs labeled with quantum dots and green fiuorescein do not significantly differ in survivin expression or the transfected cell proliferation and apoptosis rate, but quantum dot labeling can be more stable with longer maintcnance of the labeling.

Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Fluorescein ; chemistry ; Gene Expression ; Humans ; Inhibitor of Apoptosis Proteins ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; chemistry ; genetics ; Quantum Dots ; Reverse Transcriptase Polymerase Chain Reaction ; Staining and Labeling ; methods ; Transfection

OBJECTIVETo evaluate the specific killing effects of combination of recombinant adenovirus mediated double suicide gene driven by KDR promoter and survivin antisense oligonucleotide(ASODN) on colorectal cancer cells and vascular endothelial cells.

METHODSThe 293 packaging cells were transfected with the plasmids of pAdEasy-CDglyTK and the recombinant adenovirus were generated. The KDR expressive cells of SW620, ECV304 were infected with adenovirus, meanwhile survivinASODN was transferred into the same cells. The infection rate of adenovirus and transfection efficiency of survivinASODN were observed and the expression of CDglyTK was detected by RT-PCR. The expression of survivin was measured by Western blot. The killing effects and bystander effects on SW620, ECV304 were examined through MTT method.

RESULTSThe cells which were infected with the adenovirus mediated double suicide gene could be transfected with the survivin ASODN and the infection rate was not affected as well as the transfection efficiency. The high expression of CDglyTK gene was found in SW620, ECV304 cells infected with recombinant adenovirus and survivin ASODN decreased the survivin protein level. The survival rate of gene therapy group was significantly lower than that of negative group. The combination of survivin ASODN and AdKDR-CDglyTK gene therapy showed significantly lower survival rate of SW620 and ECV304 cells as compared with the AdKDR-CDglyTK or survivin ASODN used alone (P<0.05). The survival rate was slightly lower in GCV 100 microg/ml, 5-FC 2000 microg/ml than that AdKDR-CDglyTK used alone (P>0.05). The combined therapy of AdKDR-CDglyTK and survivin ASODN showed synergistic killing efficacy and more significant bystander effects.

CONCLUSIONThe combined gene therapy of AdKDR-CDglyTK system and survivin ASODN has stronger specific killing effects on colorectal cancer cells and vein endothelial cells.

Adenoviridae ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; Endothelial Cells ; metabolism ; Genes, Transgenic, Suicide ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Receptors, Vascular Endothelial Growth Factor ; genetics ; Transcription Initiation Site

OBJECTIVETo evaluate the feasibility of multi-slice spiral CT scan to localize upper airway stricture in patients with obstructive sleep apnea syndrome (OSAS) during drug-induced sleeping.

METHODSOne hundred and fourteen patients diagnosed as OSAS by polysomnography were included in the study. Multi-slice spiral CT scan covering upper airway was performed at the end of inspiration and clear upper airway images were obtained in waking. After injecting 5 mg of midazolam intravenously slowly in 109 patients, CT scan was performed at apnea and clear upper airway images were obtained in sleeping. Cross-section area and minimal diameter of airway were measured and the parameters were compared under those two states. Upper airway was displayed intuitionisticly by using post-processing techniques.

RESULTSOne hundred and nine patients with OSAS finished the examination with a success rate of 100 %. Airway obstruction at retropalatal level was observed in 62 patients, among whom 26 were associated with airway obstruction at retroglossal level, 27 with narrower airway at retroglossal level in sleeping compared with that in waking, and 9 with no significant change of the airway at retroglossal level after sleeping. Narrower airway at retropalatal level in sleeping compared with that in waking was observed in 40 patients, among whom 20 were associated with narrower airway at retroglossal level in sleeping compared with that in waking, 10 with complete airway obstruction at retroglossal level in sleeping, and 7 with no significant change of the airway at both retropalatal and retroglossal levels before and after sleeping. Minimal mean cross-section area of airway at retropalatal level was (72.60 +/-45.15)mm(2) in waking and (8.26 +/-18.16)mm(2) in sleeping; and minimal mean cross-section area of airway at retroglossal level was (133.21 +/-120.36)mm(2)in waking and (16.73 +/-30.21)mm(2) in sleeping (P <0.01). Minimal mean diameter of airway at retropalatal level was (6.91 +/-2.23) mm in waking and (1.18 +/-2.14) mm in sleeping; and minimal mean diameter of airway at retroglossal level was (8.68 +/-4.32) mm in waking and (1.68 +/-2.22) mm in sleeping (P <0.01).

CONCLUSIONMulti-slice spiral CT with post-processing techniques can display the shape of the upper airway in patients with OSAS in sleeping, and can localize the upper airway stricture and assess its range accurately.

Adult ; Aged ; Airway Obstruction ; diagnostic imaging ; Female ; Humans ; Hypnotics and Sedatives ; administration & dosage ; Male ; Middle Aged ; Oropharynx ; physiopathology ; Palate, Soft ; physiopathology ; Sleep Apnea, Obstructive ; diagnostic imaging ; Tomography, Spiral Computed ; Young Adult

Bone defects,a challenge in orthopedics,are mostly caused by severe trauma,bone tumor and bone infection.Their chief treatments include bone graft,distraction osteogenesis and Masquelet technique.Compared with other treatments,Masquelet technique is simpler and causes fewer complications,especially for infectious defects.In the first stage of Masquelet technique,a bone defect is filled with bone cement.The spacer most commonly used is polymethylmethacrylate(PMMA).PMMA can form induced membrane rich in blood vessels and bioactive components promoting osteogenesis,but it is not biodegradable,lacks bone conductibility,has an obvious thermal effect during operation,and is hard to remove in the second phase of Masquelet technique.Calcium sulfate and calcium phosphate bone cement are biodegradable materials which overcome the defects of PMMA due to their biological absorbability,bone conduction and bone inducement.It is possible for them to replace PMMA in Masquelet technique,but their poor mechanical strength limits their application.There is clinical evidence showing that calcium sulfate bone cement can form obvious induced membrane.As calcium silicate cement can also form a layer of fibrous tissue around the shell,it may be applied in orthopaedic surgery after improvement as a potential filling agent.A structure like induced membrane can form around a silicone prosthesis,and it functions similarly as induced membrane around PMMA and is more resistant to a radiation environment.Silicone has obvious advantages over PMMA after resection of bone tumor.Spacers used in the first stage of Masquelet technique have important effects on formation process and quality of induced membranes.By comparing membranes induced by different types of spacer,we can know more about the effects of spacers on induced membranes in the first stage of Masquelet technique and thus discover the most suitable spacers good for fracture healing and membrane formation.

OBJECTIVETo evaluate the selectively killing effects of combination of adenovirus mediated double suicide gene driven by kinase-domain insert containing receptor (KDR) promoter and survivin antisense oligonucleotide on breast tumor cells and vein endothelial cells.

METHODSHuman embryonal kidney cells 293 were transfected with the plasmids of pAdEasy-KDR-CDglyTK and the adenovirus was generated. The KDR expressive cells of MCF-7, ECV304 were infected by adenovirus and survivin ASODN was transferred into the same cells meanwhile. The infection rates of adenovirus and transfection efficiency of survivin ASODN were observed and the expression of CDglyTK was detected by RT-PCR. The expression of survivin was measured by Western blot. The killing effects and bystander effects on cells were assessed by MTT assay.

RESULTSThe cells infected by the adenovirus mediated double suicide gene could be transfected with the survivin ASODN and the infection rate was not affected as well as the transfection rate. The high expression of CDglyTK gene was found in the two kinds of cells and survivin ASODN decreased the survivin protein level. When survivin ASODN was transferred into MCF-7, ECV304 cells, the survival rates were 56.4% +/- 3.8% and 55.9% +/- 3.6% respectively. The combination of survivin ASODN and AdKDR-CDglyTK gene therapy showed significantly lower survival rate comparing with using each treatment alone (P < 0.05) and the survival rate decreased gradually with the increasing of the concentration of GCV and 5-FC. But the survival rate for combined gene therapy group was slightly lower in GCV 100 microg/ml, 5-FC 2000 microg/ml than that of AdKDR-CDglyTK group (P > 0.05). The combination of survivin ASODN and AdKDR-CDglyTK therapy showed synergistic killing efficacy and more significant bystander effects.

CONCLUSIONThe combined therapy with AdKDR-CDglyTK system and survivin ASODN shows obvious killing effects on breast tumor cells and vein endothelial cells.

Adenoviridae ; genetics ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Cell Survival ; Endothelial Cells ; metabolism ; pathology ; Female ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; methods ; Genetic Vectors ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; Promoter Regions, Genetic ; Receptors, Vascular Endothelial Growth Factor ; genetics ; Transfection