OBJECTIVETo investigate the silencing effect of gene encoding peroxisome proliferator-activated receptor gamma (PPARgamma) on the expression of tumor necrosis factor alpha (TNFalpha) by constructing vectors for RNA interference in RAW264.7 cells.

METHODSThe pSUPER-EGFP vectors were used to transcribe functional small interfering RNA (siRNA). Four pairs of oligonucleotides (64 nt) targeting PPARgamma gene were inserted into the downstream of the H1 promotor, with their veracity confirmed by double digestion and sequencing. Western blotting and immunofluorescence assay were used to examine the silencing effect of PPARgamma gene in RAW264.7 cells. Meanwhile, the TNFalphalevel was determined by Sandwich ELISA.

RESULTSCompared with other recombinant pSUPER-EGFP vectors (R-pSUPER.EGFP), R-pSUPER.EGFP2 induced the best silencing effect on the expression of PPARgamma in RAW264.7 cells, which played an obvious inhibitory role in down-regulating the TNFalphaexpression after the curcumin and lipopolysaccharide (LPS) stimulation.

CONCLUSIONSPPARgamma-pSUPER-EGFP inducing a silencing effect on the expression of PPARgamma can efficiently play a negative role in controlling the inflammatory responses of RAW264.7 cells.

OBJECTIVETo investigate the role of N-cadherin and fibronectin during chondrogenesis.

METHODSImmunohistochemical method and autibody induced changes of aggregation of cells were used to assay the expressions of N-cadherin and fibronectin during cell differentiation.

RESULTSThe N-cadherin was present in the area of the cell nodular area in the 24 hours group after adding chondrogenic revulsant, then there was a down-regulating trend. Fibronectin was expressed in 48 and 72 hours groups after adding chondrogenic revulsant, and showed to be negative afterward. The antibody against fibronectin or N-cadherin could inhibit the formation of cellular nodule markedly.

CONCLUSIONSCell adhesion factors play an important role during cell differentiation. TGF-beta(1) stimulates chondrogenesis via transition from an initial N-cadherin-contributing stage to a succedent fibronectin-contributing stage during the process of chondrogenesis in MSCs. Further study is needed to evaluate whether or not it can promote chondrogenesis by transfecting cDNA of CAMs to MSCs.

OBJECTIVETo evaluate current diagnosis and therapeutic effect and outcome of diffuse axonal injury (DAI) in 169 patients.

METHODSThe data of 169 DAI patients treated in the Second, Sixth, Eighth and Ninth Hospitals of Shenzhen and Shekou Hospital from January 2001 to January 2005 were collected. The imaging features, classification, GCS (Glasgow coma scale), treatment and outcome of the 169 patients were retrospectively analyzed.

RESULTSThe simpler the imaging features, the closer the focus of DAI to the periphery of hemisphere and the higher the GCS score, the better the prognoses of DAI patients will be.

CONCLUSIONSThe prognoses of DAI patients are closely related to the imaging features and classification, GCS and clinical treatment.

OBJECTIVETo study whether human neural progenitor cells can differentiate into neural cells in vivo and improve the recovery of injured spinal cord in rats.

METHODSHuman neural progenitor cells were transplanted into the injured spinal cord and the functional recovery of the rats with spinal cord contusion injury was evaluated with Basso-Beattie-Bresnahan (BBB) locomotor scale and motor evoked potentials. Additionally, the differentiation of human neural progenitor cells was shown by immunocytochemistry.

RESULTSHuman neural progenitor cells developed into functional cells in the injured spinal cord and improved the recovery of injured spinal cord in both locomotor scores and electrophysiological parameters in rats.

CONCLUSIONSHuman neural progenitor cells can treat injured spinal cord, which may provide a new cell source for research of clinical application.

OBJECTIVETo investigate the expression characteristics of basic fibroblast growth factor (bFGF) and its receptors, flg (FGFR1) and bek (FGFR2), in fetal skin at different gestational ages underlying the relevance of these 3 proteins to skin development and the mechanisms underlying the phenotypic transition from scarless to scar-forming healing.

METHODSEighteen specimens of fetal skin biopsies of human embryo were obtained from spontaneous abortions at different gestational ages of 13-32 weeks. Gene expression of bFGF, bek and flg was examined with reverse transcription-polymerase chain reaction (RT-PCR). The dynamic expression and distribution of these 3 proteins were detected with streptavidin peroxidase (SP) immunohistochemical staining method.

RESULTSIn the early gestational fetal skin, genes of bFGF and flg were strongly expressed and more protein contents of these 2 proteins were found as compared with the genes at late gestation fetal skin (2.446+/-0.116 and 2.066+/-0.152 versus 2.157+/-0.101 and 1.818+/-0.086, respectively, P<0.05). On the contrary, the levels of gene expression and protein content of bek were not differently expressed in the early gestational fetal skin versus the late ones. Protein particles of bFGF were mainly distributed in the epidermal cells and some fibroblasts. Bek was mainly located in the cell membrane and cytoplasm of epidermal cells while flg protein was principally located in the epidermal cells, endothelial cells and some fibroblasts.

CONCLUSIONSThe endogenous bFGF and their receptors might be involved in the cutaneous development at fetal stage. The differently expressing levels of bFGF and flg during gestation may be related to scarless or scar-forming repair during gestation.

OBJECTIVETo analyze retrospectively the surgical treatment of 21 cases of infected arterial injuries of the major limb hospitalized from 1989 to 2003 in our department.

METHODSAfter a radical debridement and drainage, an autologous vessel bypass reconstruction, simple vessel ligation or amputation were carried out respectively according to each patient's condition. In order to make a better milieu for the grafts, local musculo-cutaneous flaps were translocated to cover the wounds in 9 cases.

RESULTS1 case (4.8%) died during the treatment. 5 cases (5/21, 23.8%) underwent amputations. Blood supply reconstruction was successful in 15 cases (15/21, 71.4%). 93.3% (14/15) of the cases were successful in getting an unobstructed vessel at early stage. Thrombosis occurred in 1 case (1/15, 6.7%) and a sound result was obtained after a second operation.

CONCLUSIONSRadical debridement combined with bypass vessel reconstruction with autologous vein and with musculo-cutaneous flap translocation is an effective treatment for infective limb vascular injuries.

OBJECTIVETo investigate the effect of 360-degree retinectomy for severe ocular rupture and evaluate the related factors associated with prognosis.

METHODSA retrospective analysis was performed on 28 consecutive patients, 18 men and 10 women, with severe ocular rupture admitted to the Chinese PLA General Hospital between January 1999 and January 2002. The patients' ages ranged from 16 to 58 years, mean 29.5 years. All operations were performed by the first author. All patients underwent standard 3-port pars plana vitrectomy with scleral encircling buckle, endophotocoagulation, and tamponaded with silicone oil. Lensectomy were performed on 15 eyes. 360-degree retinotomy and retinectomy were performed at anterior equator on 16 eyes, and posterior equator on 12 eyes.

RESULTSDuring operation, all patients were confirmed to have vitreous hemorrhage and retinal detachment, in which hemorrhagic ciliary body detachment was found in 9 eyes, suprachoroidal hemorrhage in 12 eyes, retinal twisting like flower stalk in 11 eyes, and retinal incarceration in 17 eyes. Postoperative follow-up was conducted in 26 cases, ranged from 6 months to 46 months. Retinal reattachment achieved in 20 eyes (76.9%), in which three patients' retina was reattached after revitrectomy. The silicone oil was removed at the fourth month in 8 eyes postoperatively. Corrected visual acuity of 0.02 or more was in 14 eyes (70.0%) among the 20 patients with reattached retina. Visual acuity of 0.05 or more was in 7 eyes (35.0å), and 0.1 or more was in 3 eyes. The best visual acuity was 0.3 in 1 eye.

CONCLUSIONSRetinectomy may improve the prognosis of severe ocular rupture and save the visual function of some patients.

OBJECTIVETo determine the effect of albumin administration on lung injury in traumatic/hemorrhagic shock (T/HS) rats.

METHODSForty-eight adult Sprague-Dawley rats were divided into three groups randomly (n=16 in each group): Group A, Group B, Group C. In Group A, rats underwent laparotomy without shock. In Group B, rats undergoing T/HS were resuscitated with their blood plus lactated Ringer's (twice the volume of shed blood). In Group C, rats undergoing T/HS were resuscitated with their shed blood plus additional 3 ml of 5% human albumin. The expression of polymorphonuclear neutrophils CD18/CD11b in jugular vein blood was evaluated. The main lung injury indexes (the activity of myeloperoxidase and lung injury score) were measured.

RESULTSSignificant differences of the expression of CD18/11b and the severity degree of lung injury were founded between the three groups. (P<0.05). The expression of CD18/CD11b and the main lung injury indexes in Group B and Group C increased significantly compared with those in Group A (P<0.05). At the same time, the expression of CD18/CD11b and the main lung injury indexes in Group C decreased dramatically, compared those in Group B (P<0.05).

CONCLUSIONSThe infusion of albumin during resuscitation period can protect lungs from injury and decrease the expression of CD18/CD11b in T/HS rats.

Albumins ; therapeutic use ; Animals ; CD11b Antigen ; metabolism ; CD18 Antigens ; metabolism ; Disease Models, Animal ; Neutrophils ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; drug therapy ; etiology ; metabolism ; Shock, Hemorrhagic ; complications ; metabolism ; Treatment Outcome ; Wounds and Injuries ; complications ; metabolism

OBJECTIVETo investigate the surgical treatment methods and evaluate the outcome of gunshot wounds combining with seawater immersion in rabbits' femoral arteries.

METHODSOne hundred healthy New Zealand white rabbits (either sex, 3.14 kg+/-0.61 kg in weight) were randomly divided into a seawater immersion group (n=50) and a simple injury group (n=50). The unilateral femoral arteries of all the rabbits were injured by 0.38 g steel spheres with velocity of 600-800 m/s fired by a 7.62 mm rifle. The rabbits in the seawater immersion group were immersed in seawater (saline content of 2.54%, pH 8.2-8.4, and at 21 degrees C) for 60 minutes but those in the simple injury group were not. After the injured segment (observed by naked eyes) of the femoral artery was excised, the blood flow restoration was reconstructed by direct end-to-end anastomosis, reversed autogenous venous grafting or cryopreserved arterial allografting, according to the length of the arterial defects. At 24 hours, and 7, 14 and 21 days after operation, the blood flow was examined. Operative exploration was performed for the animals with partly or fully obstructed blood flow. The tissues around the anastomosis sites and the grafts were harvested for pathological observation under a light microscope and an electron microscope.

RESULTSIn the rabbits with completely transected injury, the unobstructed rates in the first 3 weeks after operation were 80.00% in the seawater immersion group and 86.67% in the single injury group, and no significant difference was found between the two groups (P>0.1). In the rabbits with arterial contusion injury, the unobstructed rates in the first 3 weeks after operation were 86.67% in the seawater immersion group and 82.35% in the single injury group, and no significant difference was found between the two groups (P>0.1). Most thrombosis occurred in the first operative week. Atypical endothelial cells were detected at the anastomosis sites at the first operative week, and the anastomosis sites were lined with endothelial cells in 3 weeks postoperatively.

CONCLUSIONSDuring the surgical treatment for gunshot wounds combining with seawater immersion, resection of the grossly-injured artery and routine artery reconstruction can obtain satisfactory outcome. Homologous artery is a kind of vascular graft with certain applied value.

Animals ; Disease Models, Animal ; Female ; Femoral Artery ; diagnostic imaging ; injuries ; pathology ; surgery ; Hydrotherapy ; methods ; Male ; Rabbits ; Radiography ; Random Allocation ; Seawater ; Treatment Outcome ; Wound Healing ; Wounds, Gunshot ; diagnostic imaging ; pathology ; therapy

OBJECTIVETo culture olfactory ensheathing cells (OECs) of rats in vitro and to investigate its morphology, mitosis and immunocytochemistry, and to explore if the OECs could be a new donation for transplantation.

METHODSOECs were harvested from olfactory mucosa of Sprague Dawley rats based on the differing rates of attachment of the various cell types, followed by glial fibrillary acidic protein (GFAP), nerve growth factor (NGF), anti-low affinity receptor for NGF (NGFRp75), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and S-100 immunocytochemistry. The morphological changes and mitosis were observed under a phase contrast microscope at different culture time.

RESULTSThree morphologically distinct types of cells, bipolar, multipolar and flat morphology were present in the primary culture of adult rat olfactory mucosa. Mitosis was characterized by a retraction of all processes, forming a sphere that divided into spherical daughter cells, the daughter cells sent out their processes. The OECs were immunoreactive for GFAP, NGFRp75, S-100, NGF, BDNF and NT-3.

CONCLUSIONSThe OECs from nasal olfactory mucosa cultivated in the medium with fetal bovine serum could survive, divide, differentiate, and express the neurotrophin. It may become an accessible source for autologous grafting in spinal cord injury.

Animals ; Cells, Cultured ; Disease Models, Animal ; Immunohistochemistry ; Male ; Mitosis ; Olfactory Mucosa ; cytology ; transplantation ; Rats ; Rats, Sprague-Dawley