This study was designed to creat a new method for detecting the degree of cross-linked allogenic tissue based on fluorescent technique. The thoracic aorta of New Zealand rabbits were divided randomly into four groups according to the concentration of Glutaraldehyde (GA), which were group A (control group-with no GA), group B (cross-linked with GA of 0.625%), group C (1.25%), and group D (2.5%). Each group was cross-linked with GA and reacted with 5-FAMSE, and then the fluorescence intensity was observed via fluorescence microscopy (analyzed with Image-Pro Plus 6.0, a professional image analysis software). The differences between groups in order of fluorescence intensity were found to be: group A > roup B > group C > group D (P < 0.01). Meanwhile, the tissue proteins extracted from aorta in each group were submitted to conventional polyacrylamide gel electrophoresis (PGE) after being cross-linked with GA; the result from this method was compared with that from the method of 5-FAMSE. In group A, the tissue proteins extracted from the aorta cross-linked with GA were obviously less than those not cross-linked with GA. However, this phenomenom was not clearly seen among the B, C and D groups. Nevertheless, 5-FAMSE can detect the degree of cross-linkage more conveniently and directly.
Animals ; Aorta, Thoracic ; drug effects ; transplantation ; Bioprosthesis ; Blood Vessel Prosthesis ; Cross-Linking Reagents ; pharmacology ; Esters ; Female ; Fluoresceins ; Fluorescence ; Glutaral ; pharmacology ; Male ; Microscopy, Fluorescence ; Rabbits

AIM:To generate thalassemia-specific integration-free induced pluripotent stem cells ( iPSC) and to detect their ability of differentiation into hematopoietic precursors .METHODS:The plasmids pEB-C5 and pEB-Tg were transfected into the fibroblast cells from hemoglobin Bart ’ s hydrops fetalis ’ s skin by the method of nuclear transfection to reprogramm the cells into iPSC .The ability of the iPSC to differentiate into 3-germ layer cells was determined .The iPSC were cocultured with mouse OP 9 cells to differentiate into hematopoietic precursors and the hematopoietic precursor specific antigens were detected .RESULTS:The integration-free iPSC from hemoglobin Bart ’ s hydrops fetalis ’ s skin fibroblasts were successfully derived, and had the ability to differentiate into 3 germ layers.When cocultured with OP9 cells for 9 d, the positive rate of hematopoietic progenitor cell marker CD 34 was 18.7%, and the CD34 and CD45 double positive rate was 12.2%.CONCLUSION:Hemoglobin Bart ’ s hydrops fetalis ’ s skin fibroblasts can be successfully induced into “in-tegration-free” iPSC.This cell line has the ability to differentiate into 3 germ layers , and can be differentiated into hemato-poietic precursors when cocultured with OP 9 cells.

Objective To dynamically observe stenosis and wall thickness of carotid artery with endothelium injury in mouse using 7T micro-MR imaging in vivo. Methods A mouse model of carotid artery intimal injury was established by removing endothelium with a flexible wire. The lumen diameter, lumen area, wall thickness and wall area of the injured arteries were observed, and serial MR scanning was performed in different time points after operation. Results The injured arteries and perivascular parenchyma were clearly observed by MR imaging. Before and 1, 5, 10 and 15 days after artery injury, the lumen diameter were (0.57±0.07)mm,(0.41±0.19)mm, (0.44±0.10)mm, (0.43±0.10)mm and (0.47±0.11)mm respectively, and the lumen area were (0.30±0.06)mm2, (0.18±0.11)mm2, (0.18±0.06)mm2, (0.18±0.06)mm2 and (0.22±0.07)mm2. The thickness of artery wall was(0.23±0.12)mm, and the area of artery wall was (0.35±0.24)mm2 15days after artery injury. Conclusions Stenosis and wall thickening of carotid artery after the artery intimal injury of mouse can be dynamically observed on MR imaging in vivo.

Objective To study the effects of water decoction and chloroform extraction fraction before and after processing of Aconitum sinomontanum Nakai (ASN) on the analgesic and anti-inflammatoryin mice.Methods Kunming mice were randomly divided into 10 groups (10 mice in each group)according to the weight:blank control group (0.5％ methylcellulose),positive control group (10 mg · kg-1 indomethacin + 1.6 mg · kg-1 lappaconitine),the high and low dose groups of water decoction of raw ASN (0.9,0.3 g · kg-1),the high and low dose groups of water decoction processed product (0.9,0.3 g · kg-1),the high and low dose groups of chloroform extraction fraction of ASN (4.5,1.5 mg · kg-1),the high and low dose groups of chloroform extraction fraction of processed product (4.5,1.5 mg · kg-1).Hot plate was used to study the analgesic effect of water decoction and chloroform extraction fraction before and after processing of ASN.Auricular swelling induced by dimethylbenzene in mice were observed for the evaluation of anti-inflammatory effects of it.Results Analgesic activity:Inthe hot plate test,the pain threshold of high dose groups of water decoction of processed product at 30 minafter the last administration was (24.1 ± 6.3) s,the pain threshold of high dose groups of water decoction of processed product,high and low dose groups of chloroform extraction fraction of raw product,high dose groups of chloroform extraction fraction of processed product at 120 min after the last administration were(49.7 ± 9.7),(49.8 ±7.4),(47.3 ± 6.3),(45.1 ± 7.1)s,compared with blank control group,the difference were significant (all P ＜ 0.05),respectively.Anti-inflammatory activity:The swelling degree of high dose groups of water decoction of raw product,high and low dose groups of water decoction processed product,high and low dose groups of chloroform extraction fraction of raw and processed product were (16.9 ± 11.2),(15.2 ±4.8),(17.7 ±6.4),(14.2 ±4.2),(16.0 ± 4.5),(15.7 ± 5.2),(12.5 ± 5.5) mg,compared with blank control group,the difference were significant (all P ＜0.01).The each groupof processed products compared with the each group of raw product,the difference was not significant (P ＞ 0.05).Conclusion The raw Aconitum sinomontanum Nakai have strong analgesic and anti-inflammatory effects,and processed product still remain strong analgesic and anti-inflammatory effects,and have a trend to increase compared with the raw product.

Objective To explore the applied value of electroencephalogram (EEG) in assessment of psychiatric impairment among patients with mental disorders due to traumatic brain injury.Methods According to the ICD-10,a total of 271 subjects were enrolled and assessed with the criterion of mental disorders due to traumatic brain injury.Activity of Daily Living Scale (ADL),Functional Activities Questionnaire (FAQ) and Social Disability Screening Schedule (SDSS) were used to evaluate the severity of patients.All the participants were tested by Wechsler Adult Intelligence Scale (WAIS) and examined by EEG.Results Totally 215 patients accomplished the study.The results of Glasgow Coma Scale (GCS),the severity of craniocerebral injury and the scores of FAQ,SDSS and ADL showed significant difference among the patients with different severity of EEG (P＜0.05).The grades of psychiatric impairment showed significant difference among the patients with different abnormal EEG (P＜0.05).Conclusion EEG can reflect the severity of craniocerebral injury,assist evaluate the social function and activity of daily living of patients with mental disorders due to traumatic brain injury,and distinguish the mild psychiatric impairment grades,which suggest that EEG has a certain reference value in the assessment of psychiatric impairment.