1.Apoptotic Change and NOS Activity in the Experimental Animal Dif fuse Axonal Injury Model.
Yonsei Medical Journal 2001;42(5):518-526
Although nitric oxide (NO) plays an important role in the pathophysiological process of cerebral ischemia or severe traumatic brain injury, its contribution to the pathogenesis of moderate diffuse axonal injury (mDAI) remains to be clarified. The alterations in nitric oxide synthase (NOS) activity and the histopathological response after mDAI was investigated. Forty anesthetized Sprague-Dawley adult rats were injured with a Marmarou's weight-drop device through a Plexiglas guide tube. These rats were divided into 8 groups (control, 1 hr, 2 hr, 3 hr, 6 hr, 12 hr, 24 hr, 48 hr after trauma). The temporal pattern of apoptosis in the adult rat brain after mDAI was characterized using TUNEL histochemistry. In addition, the cDNA for NOS activity was amplified using RT-PCR. The PCR products were electrophoresed on a 2% agarose gel. eNOS activity was not detected, but nNOS activity was expressed after 3 hr and continuously 48 hr after impact, which was approximately double that of the control group at 12 and 24 hr. Subsequently, there was a decrease in activity after 48 hr. The iNOS activity increased dramatically after 12 hr and was constant for a further 12 hr followed by a dramatic decrease below the level of the control group. Significant apoptotic changes occurred 12 and 24 hr. after insult. nNOS and iNOS activity were affected after moderate diffuse axonal injury in a time-dependent manner and there was a close relation between the apoptotic changes and NOS activity. Although the nNOS activity was expressed early, its activity was not stronger th an iNOS, which was expressed later.
Animal
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*Apoptosis
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Craniocerebral Trauma/enzymology/*physiopathology
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Diffuse Axonal Injury/enzymology/*physiopathology
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Nitric-Oxide Synthase/*metabolism
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Rats
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Rats, Sprague-Dawley
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Wounds, Nonpenetrating/enzymology/*physiopathology
2.A study on the expression of VEGF in the blunt injury of rat's skin.
Mei LI ; Pei-jun HUANG ; Min LIU
Journal of Forensic Medicine 2003;19(1):13-15
OBJECTIVE:
To evaluate changes of VEGF in skin after blunt injury.
METHODS:
The rats of injury groups were subjected to skin blunt injury by free-falling iron hammer. The samples taken at 1 h, 3 h, 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 9 d after injury were studied by immunohistochemistry and MIAS image analysis system.
RESULTS:
In the skin of normal control group the expression level of VEGF was low. The increase of VEGF could be observed at 1 day after injury, reached peak at 7 days and declined at 9 days.
CONCLUSION
Blunt injury in the skin could induce the expression of the VEGF. Moreover, the change pattern of VEGF level was quite regular with time and could be used to estimate the time after skin injury.
Animals
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Endothelial Growth Factors/metabolism*
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Female
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Forensic Medicine
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Intercellular Signaling Peptides and Proteins/metabolism*
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Lymphokines/metabolism*
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Male
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Rats
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Rats, Sprague-Dawley
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Skin/metabolism*
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Time Factors
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Vascular Endothelial Growth Factor A
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Vascular Endothelial Growth Factors
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Wounds, Nonpenetrating/metabolism*
3.Preliminary application on test paper of gold-labelled antibody against fibronectin EIIIA splicing variant.
Ning-Guo LIU ; Yi-Jiu CHEN ; Ren-Quan LU ; Xiao-Hua HUANG ; Jian-Hua ZHANG
Journal of Forensic Medicine 2006;22(2):81-83
OBJECTIVE:
To develop the test Paper of gold-labelled antibody against fibronectin EIIIR A splicing variant, which can be available in forensic wound interval estimation.
METHODS:
Two sensitive antibodies were compared with enzyme link immunoabsorband assay (ELISA). After colloid gold labeled, the effects of the two antibodies were tested by methods of Dot immunogold filtration assay and gold immunochromatography assay, respectively. The test paper was finally appraised by applied in experimental skin injury in rats.
RESULTS:
On the test paper, detected line appeared in three hours wound age group, and then the darkness of positive staining became darker with injury time prolonging, while control normal skin cannot found to be positive staining.
CONCLUSION
The gold-labeled test paper can be useful in estimation of wounding interval in forensic science.
Alternative Splicing
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Animals
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Antibodies, Monoclonal
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Enzyme-Linked Immunosorbent Assay
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Fibronectins/metabolism*
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Forensic Pathology
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Gold Colloid
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Immunoassay/methods*
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Reagent Strips
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Skin/metabolism*
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Time Factors
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Wounds, Nonpenetrating/metabolism*
4.Effects of penehyclidine hydrochloride on apoptosis of lung tissues in rats with traumatic acute lung injury.
Ling-li WANG ; Li-ying ZHAN ; Xiao-jing WU ; Zhong-yuan XIA
Chinese Journal of Traumatology 2010;13(1):15-19
OBJECTIVETo investigate the effects of penehyclidine hydrochloride on apoptosis of lung tissue cells and its mechanism in acute lung injury following blunt chest trauma in rats.
METHODSSprague Dawley (SD) rats (n equal to 54) weighing (250+/-5) g were divided equally and randomly into three groups: normal control group (C group, n equal to 18), trauma model group (T group, n equal to 18) and penehyclidine hydrochloride treatment group (P group, n equal to 18). Each group was further divided into three subgroups according to the time points of 3, 12 and 24 hours after experiment (at each time point, n equal to 6 for each subgroup). Rats of P group were intraperitoneally injected with penehyclidine hydrochloride for 2 mg/kg immediately after blunt chest trauma and rats in its 24 hours subgroup were once again injected with penehyclidine hydrochloride in the same dose 12 hours after injury. Lung tissue samples were collected at every time point and cell apoptosis in lung tissues were measured by TUNEL. Apoptotic index (AI) was calculated, expressions of bax and bcl-2 were detected by immunohistochemical staining of SABC, and lung tissue sections were taken for light and electron microscopic observation.
RESULTSAs compared with C group, at every time point, AI and expressions of bax and bcl-2 in T group were higher (P less than 0.05), and the ratio of bcl-2/bax markedly decreased (P less than 0.05), especially in the 24 hours subgroup. The ratio in T group (0.468+/-0.007) was lower than that in C group (1.382+/-0.058, t equal to 12.5, P less than 0.01). Lung tissue injuries were significant under a light microscope, and the number of apoptotic cells increased obviously under a transmission electron microscope. As compared with T group at the same phase, AI and expression of bax decreased in P group (P less than 0.05 and P less than 0.01), while the expression of bcl-2 increased significantly (P less than 0.01), and the ratio of bcl-2/bax markedly increased (P less than 0.05), especially in the 24 hours subgroup. The ratio in P group (1.012+/-0.070) was much higher than that in T group (0.468+/-0.007, t equal to 8.3, P less than 0.01). The injury of lung tissues was relieved, and apoptosis of cells decreased obviously under a transmission electron microscopic observation.
CONCLUSIONSApoptosis and expressions of bax and bcl-2 in lung tissues might be involved in the pathogenesis of lung injury induced by blunt chest trauma. Penehyclidine hydrochloride can alleviate lung injuries by inhibiting apoptosis of lung tissue cells, during which effects of penehyclidine hydrochloride on regulating expressions of bax and bcl-2 may play an important role.
Acute Lung Injury ; drug therapy ; metabolism ; pathology ; Animals ; Apoptosis ; drug effects ; Lung ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Quinuclidines ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Thoracic Injuries ; complications ; Wounds, Nonpenetrating ; complications ; bcl-2-Associated X Protein ; analysis