· AIM: To investigate the effect of β1-integrin overexpression on the apoptosis of rabbit corneal epithelial cells and the related mechanism. · METHODS: The plasmid expressing β1-integrin-GFP fusion protein was constructed by polymerase chain reaction (PCR), and this plasmid (β1 group) or the empty vector (mock group) was transfected into rabbit corneal epithelial cells, respectively. The expression of β1-integrin-GFP fusion gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The adhesion of transfected cells to extracellular matrix (ECM) proteins was determined by adhesion assay. The apoptosis of rabbit corneal epithelial cells was assayed by Hoechst 33342 staining and DNA ladder. The phosphorylation of mitogen-activated protein (MAP) kinase was examined by Western blot. · RESULTS: Rabbit corneal epithelial cells overexpressing β1 -integrin-GFP fusion gene were successfully established. Compared with mock group, β1-integrin transfection significantly promoted the adhesive of rabbit corneal epithelial cells to ECM proteins such as laminin, fibronectin, collagen Ⅰ and collagen Ⅳ. Β1-integrin overexpression inhibited apoptosis and induced MAP kinase phosphorylation in rabbit corneal epithelial cells (P<0.05).· CONCLUSION: These data suggest that overexpression of β1-integrin confers resistance to apoptosis in rabbit corneal epithelial cells, and MAP kinase pathway may play an important role in this process.

Objective To explore the reliability of relevant electrocardiogram(ECG) indexes in evaluating isoprenaline(ISO)-induced rat acute ischemic myocardial injury and provide reference for future scientific applications of these models. Methods Seventy male Wistar rats were randomly equally assigned to ten groups according to their body weight: 5,10,20,40,80,160,320,640, 1280 and 2560 μg/kg dose groups. All rats were tail intravenously given corresponding doses of saline diluted isoprenaline according to their body weight. Standard limb Ⅰ , Ⅱ, Ⅲ-lead ECG of all rats were recorded before, immediately after and 1,24,and 72 hour after injection, respectively.Changes of heart rate, T-wave amplitude of Ⅱ -lead and Q-T interval were measured. Results Significant differences were found in heart rates, T-wave amplitudes and Q-T intervals at different time points(F = 15.03,11.28,13.64, all P ＜ 0.01 ), while differences among the ten ISO-dose groups were statistically insignificant (F= 1.45, 1.17,1.09, all P ＞ 0.05). No interaction between observation time and ISO dose was observed on heart rates, T-wave amplitudes and Q-T intervals(F= 0.79,0.82,0.59, all P ＞ 0.05). Immediately after injection of ISO, the heart rates were significantly increased compared with that of pre-injection in all groups(all P ＜ 0.05), of which 320 and 640μg/kg dose groups increased most significantly [(550 ± 47), (521 ± 43)times/min]. T-waves decreased significantly compared with that of pre-injection (all P ＜ 0.01 ), and 20 μg/kg dose and above groups decreased particularly evident, and partly inverted. Q-T intervals of rats in each group were significantly shorter than that of pre-injection(all P ＜ 0.01 ), and 320, 640, 1280 μg/kg groups shortened more pronounced[(0.070 ± 0.006),(0.072 ± 0.005), (0.068 ± 0.005)ms]. One hour after injection, the heart rate of rats in each group decreased,except 320 and 640 μg/kg dose groups[(518 ± 43), (487 ± 36)times/min], which were still higher than that of pre-treatment[(450 ± 40), (448 ± 51 )times/min, all P ＜ 0.05], the rest groups no longer had significant differences (all P ＞ 0.05). ECG T-wave in each group was significantly recovered compared with that of instantly medication (all P＜0.05), and 40 μg/kg dose and above groups recovered more than a big margin, but there were still differences compared with that of pre-treatment (P ＜0.05), while T-waves of 40 μg/kg dose and below groups had returned to the level of pre-treatment. Q-T interval in each group had varying degrees of recovery, except 1280 and 2560 μg/kg dose groups[(0.080 ± 0.004), (0.076 ± 0.011 )ms]which were still less than that of pre-treatment[(0.086 ± 0.007),(0.085 ± 0.006)ms, all P ＜ 0.05], other groups had no significant difference compared with that of pre-treatment (all P ＞ 0.05). Twenty-four hours after injection of ISO, the heart rates of 1280 and 2560 μg/kg dose groups [(389 ± 31 ), (398 ± 23)times/min]decreased significantly compared with that of pre-treatment[(427 ± 43), (438 ±26)times/min, all P ＜ 0.05], while other groups had returned to the level of pre-treatment. Seventy-two hours after injection of ISO, the heart rates, T-wave amplitudes and Q-T intervals of all doses groups had returned to the level of pre-treatment (all P ＞ 0.05). Conclusions There has no significant ST segment in the electrocardiogram of rat.Isoprenaline has an exact effect on shortening Q-T interval. T-wave amplitude and Q-T interval can be used as reliable indexes of ECG for assessment of this animal model.

Objective To prepare a kind of superparamagnetic iron oxide (SPIO) and doxorubicin loaded multifunctional polymer microbubbles (MPMBs),and to explore its potential application as an ultrasound(US)/magnetic resonance (MR) imaging contrast agent in vitro.Methods The MPMBs and normal polymer microbubbles (PMBs) were made by double emulsion and freeze-drying methods.The physical property,drug encapsulation efficiency and the drug-loading efficiency of MPMBs were determined,and the release property and US/MR imaging enhancement of MPMBs were observed.Results The MPMBs had a regular shape and narrow size distribution.The drug encapsulation efficiency was (60.20±2.69) ％,and the drug-loading efficiency was (6.02 ± 0.27) ％.The in vitro release experiment showed that ultrasound can promote the release of doxorubicin in MPMBs.US imaging in vitro showed that the enhancement of MPMBs was better than PMBs,and MR imaging in vitro conformed that MPMBs could well enhance MR imaging.Conclusions The MPMBs is a multifunctional contrast agent with the treatment function as well as US/MR dual-mode imaging enhancement effect.

Objective To prepare polyclactic-co-glycolic acid(PLGA) polymer ultrasound contrast agent loading sudan black dye (SB-PLGA) and study the enhancement effect of it on rabbit lymph nodes imaging and as a sentinel lymph node (SLN) biopsy in the feasibility and value of the indicator.Methods SB-PLGA ultrasound contrast agent was made by double emulsion and freeze-drying methods.Thigh subcutaneous rabbit VX2 tumor model of eight lymph nodes were set up with the foot pad subcutaneous injection of contrast agents to get contrast-enhanced ultrasound imaging of the popliteal lymph node and the second inguinal lymph node stations.Then the blue stained lymph nodes were resected by popliteal lymph node dissection and the second station lymph nodes were also observed after 30 mins respectively,lymph nodes were examined by frozen sections HE staining in parallel.Results SB-PLGA microbubbles had a tight size distribution.Ultrasound imaging can be significantly enhanced lymph node imaging,the second station lymph node imaging was not obvious.Popliteal lymph node dissection for lymph node staining,30minutes later,frozen sections HE staining were seen within the lymph node contrast agent present in the lymphatic sinus and a large entry of macrophages.Inguinal lymph node dissection showed the second station of lymph nodes no blue staining.Conclusions SB-PLGA ultrasound contrast agent is good for sentinel lymph node imaging and biopsy indicator.Contrast agent retention and accumulation in the lymph nodes by the uptake of macrophages may be associated with the mechanisms of lymph node enhanced imaging.

Objective To observe the curative effect, safety and patient's satisfaction degree of combining compound betamethasone with anisodamine injection in the blocking treatment of eyebrow alopecia areata.Methods A total of 66 cases were randomly divined into two groups: patients were treated by compound betamethasone ( group A) and combining compound betamethasone with anisodamine ( group B). The drugs were injected from the middle of eyebrow alopecia areata to two sides by the standard of 0. 2 ml/cm2 in both groups one time a month. The curative effect of the all cases were observed at 1, 2 and 3 months after treatment. The safety of them were evaluated based on incidence rate of the side effect and their satisfactory degree were evaluated based on subjective reception 3 months after treatment. Results The effective rates in the group B were 58.82 %, 73.53 % and 88.23 % by turns at 1,2 and 3 months after treatment, and those in the group A were 31.25 %, 53.13 % and 71.88 %, respectively. The curative effect in the group B was better than that in the group A (P ＜ 0. 05), and the incidence rate of side effect in the group B was similar to that in the group A (P＞0.05) and the patient's satisfaction degree in group B was better than that in group A (P ＜0. 05) 3months after treatment. Conclusion The compound betamethasone combined with anisodamine injection in the treatment of eyebrow alopecia areata has superior curative efficacy, more safety and higher satisfactory degree in the patients.

Objective To explore therapeutic effects and mechanisms of radical scavenger edaravone on experimental cerebral hemorrhage.Methods Two hundred-forty male SD rats were divided randomly into four groups:control group,cerebral hemorrhage group,edaravone treatment group before operation (A) and edaravone treatment group after operation (B).Experimental cerebral hemorrhage model was made according to the method reported by Rosenberg.Water quantity contained in brain and nervous missing sign were observed,meanwhile the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in brain tissue were measured.Results Compared with cerebral hemorrhage group,nervous missing sign and water quantity contained in brain obviously changed in edaravone treatment group (P

Objective To evaluate the effect of three diamension-digital subtraction angiography (3D-DSA) or computed tomography angiography (CTA) on the patients with ruptured multiple intmcranial aneurysm (MIA). Methods A retrospective study on 21 patients with MIA was performed.After scanning with 3D-DSA or 3D-CTA, three-dimensional reconstruction of MIA was carried out by 3D workstation,then the diagnosis was decided and the treatment plan (endovascular treatment or microsurgery) was selected according to stereoscopic image of MIA. Results (1) 3D-DSA or CTA was performed in 21 patients with subarachnoid hemorrhage (SAH),it was revealed these patients carried with 48 aneurysms,including 35 small aneurysms (25 mm).Not only miero-aneurysms and small aneurysms could be precisely showed,also the size of aneurysmal neck,the relationship of the aneurysm and the parent vessel and contiguous branches by stereoscopic image.(2) According to the standard of classification,9 patients with MIA for gradeⅠ(42.9%),10 for gradeⅡ(47.6%),2 for gradeⅢ(9.5%),0 for gradeⅣ.Endovascular treatment was selected prior to microsargery for those high grade patients.In this group,17 patients with 40 aneurysms underwent endovascular embolotherapy with GDC coils.Twenty four anemysms were completely occlusioned,12 beyond 90%,4 were left without treatment because of their small size.In microsurgery group,3 aneurysrus were totally clipped,1 could not be found during operation.No any treatment was accepted in 2 patients with 4 aneurysms. Conclusions 3D-DSA or CTA,which is very useful for the diagnosis and treatment of MIA,could improve the accuracy of diagnosis of MIA and clearly show the stereoscopic image of MIA,also the relation of sac and parent artery.For those patients with high grade MIA,endovascular treatment was selected prior to microsurgery,pro re nata,used to combine with mierosurgery.

[Objective]To investigate the value of"pericardium crescent sign"in the diagnosis of non-calcified con-strictive pericarditis in CT.[Methods]Twenty-seven patients with non-calcified constrictive pericarditis and forty-six cardiac tumors confirmed by surgical pathology were analyzed retrospectively. All patients underwent non-enhanced and enhanced CT scan preoperatively.Pericardial morphology,density,enhancement features,appearance of atrium,ventri-cle and inferior vena cava of non-calcified constrictive pericarditis were analyzed. Pericardial morphology of cardiac tumors was also analyzed.[Results]Of the 27 patients with non-calcified constrictive pericarditis,18 patients developed"pericardium crescent sign",accounting for 66.7%,ventricular varying degrees of deformation in 13 cases,accounting for 48%;venous dilatation in 26 cases,accounting for 93%;atrial enlargement in 7 cases,accounting for 26%. In 46 patients with cardiac tumors,only 1 patient had a similar"pericardial crescent"sign.For the diagnosis of constrictive pericarditis using pericardial crescent sign,the sensitivity was 66.7%,the specificity was 97.8%,Youden index was 0.64. The area under the ROC curve was 0.82([0.71-0.94],P<0.001).[Conclusion]"Pericardium crescent sign"is important CT features of non-calcified constrictive pericarditis. It has high specificity for differentiating non-calcified constrictive pericarditis from pericardial tumor imaging.

OBJECTIVETo observe the protection of Qingyuan Shenghua Decoction (QSD) on multiple organs of sepsis patients after bone trauma, and to preliminarily explore its mechanism.

METHODSTotally 60 sepsis patients after bone trauma were randomly assigned to the treatment group and the control group according to random digit table, 30 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took QSD or were nasally fed with QSD, one dose per day for 1 week. Changes of WBC, oxygenation index (PaO2/FiO2), serum creatinine (SCr), total bilirubin (TBIL), aspartate aminotransferase (AST), fibrinogen (FIB), D-dimer (DD), activated partial thromboplastin time (APTT), pro-calcitonin (PCT), C-reactive protein (CRP), heart rate (HR), mean arterial pressure (MAP), intra-abdominal pressure, scores for Acute Physiology and Chronic Health Evaluation II (APACHE II), sequential organ failure assessment (SOFA) scores were observed before treatment and on day 1, 3 and 7 after treatment.

RESULTSCompared with the control group at the same time point, MAP increased at post-treatment day 1 and 3; CRP, APTT, HR, SCr, TBIL, AST, intra-abdominal pressure at post-treatment day 3 obviously decreased in the treatment group (P < 0.05, P < 0.01). WBC, SOFA scores, PCT, CRP, APACHE II, APTT, D-D, HR, SCr, TBIL, AST and intra-abdominal pressure significantly decreased; FIB, MAP and PaO2/FiO2 obviously increased at post-treatment day 7 (P < 0.05, P < 0.01).

CONCLUSIONQSD had good protective effect on multiple organ function in sepsis patients after bone trauma, and its mechanism might be related with effectively clearing endotoxin, alleviating inflammatory reactions, and fighting against coagulation dysfunction.

APACHE ; Blood Coagulation ; Bone Diseases ; complications ; C-Reactive Protein ; metabolism ; Calcitonin ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Inflammation ; Partial Thromboplastin Time ; Protein Precursors ; metabolism ; Sepsis ; drug therapy ; etiology

OBJECTIVEThe multiple levels fragmentations of five furocoumarine (psoralen, xanthotoxin, bergapten, oxypeucedanin, and byakangelicol) in Angelica dahurica have been demonstrated using LTQ-Orbitrap mass spectrometry with high resolution and high mass accuracy to discover the possible,fragmentation regularity.

METHODDuringcollsion-induced dissociation (CID), the MS(n) data of the five compoundswhich were gained in the positive ion mode at 35ev collision energy by direct injection syrings method were analyzed using Xcalibar 2.0 Software to infer the formula of these fragmentations.

RESULTThe results indicated that the five compounds have similar fragmentation process with CO meutral lost at C5,C8-subsituents and furan ring, meanwhile the meutralloss of CO2 occurred easily at lactone group.

CONCLUSIONThis method is helpful in identifying the structures of other furocoumarinein Angelica dahuricaand their metabolites in vivo.

Angelica ; chemistry ; Chemical Phenomena ; Coumarins ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; methods ; Molecular Structure