Recently, activation of the adenosine A2A receptors has been shown to exert protection against peripheral tissue injuries but aggravation in the central nervous system (CNS) injuries. To explore the different effects of adenosine A2A receptors and try to perform some new treatment strategies for peripheral tissue and CNS traumas, we constructed the mouse models of skin trauma, skin combined radiation-impaired wound and traumatic brain injury (TBI), respectively. Wild type mice and A2A receptor gene knockout mice were both used in the experiments. In skin trauma and combined radiation-impaired wound models, the time of wound healing was observed, while in TBI model, neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The results showed that in skin trauma and combined radiation-impaired wound models, CGS21680 (an agonist of the A2A receptors) promoted while A2A receptor gene knockout delayed the course of skin wound healing. On the contrary, in TBI model, A2A receptor gene knockout, not CGS21680, showed a protective role by inhibition of glutamate release. These data further indicate that promoting glutamate release may account for the different effects of A2A receptor activation in CNS injury and peripheral tissue injury models. These findings may provide some experimental evidence and a new strategy for clinical treatment of peripheral tissue damages by agonists of A2A receptors, while treatment of CNS injuries by antagonists of A2A receptors.
Adenosine ; analogs & derivatives ; pharmacology ; Animals ; Brain ; pathology ; Brain Injuries ; physiopathology ; Disease Models, Animal ; Glutamic Acid ; cerebrospinal fluid ; Mice ; Mice, Knockout ; Phenethylamines ; pharmacology ; Receptor, Adenosine A2A ; genetics ; physiology ; Wound Healing

The aim of this study is to investigate the effect of (S)-4-carboxy-3-hydroxy-phenylglycine [(S)-4C3HPG], a mixed group I glutamate metabotropic receptor antagonist and a group II agonist, on impairment in a cortical impact model of traumatic brain injury (TBI) in mice and to elucidate the possible mechanisms. Mice were injected (i.p.) with saline, 1 mg/kg (S)-4C3HPG, 5 mg/kg (S)-4C3HPG and 10 mg/kg (S)-4C3HPG (n=10 per group), respectively, at 30 min before moderate TBI. Neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The expressions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) mRNA in injured cortex were also detected by real-time RT-PCR. The results showed that the neurological deficits and cerebral edema were significantly attenuated in mice pretreated with (S)-4C3HPG (5 and 10 mg/kg respectively) compared with those in mice pretreated with saline. Furthermore, (S)-4C3HPG treatment also decreased the glutamate concentration in CSF and the expressions of TNF-α and IL-1β mRNA remarkably in a dose-dependent manner. These results suggest that (S)-4C3HPG treatment attenuates cortical impact-induced brain injury possibly via suppression of glutamate release and inhibition of excessive inflammatory cytokine production. These findings highlight the potential benefit of glutamate metabotropic receptor ligand for preventing TBI.
Animals ; Brain Injuries ; drug therapy ; metabolism ; physiopathology ; Cytokines ; metabolism ; Glutamic Acid ; cerebrospinal fluid ; Glycine ; analogs & derivatives ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Receptors, Metabotropic Glutamate ; agonists ; antagonists & inhibitors

Purpose To explore the mutation of CD79B and MyD88 in primary testicular diffuse large B cell lymphoma and their significance.Methods Histopathologic features were observed in 15 cases of primary testicular diffuse large B cell lymphoma and immunophenotype was analyzed by immunohistochemical staining (IHC).Sanger sequencing was used to detect CD79B Y196 and MyD88 L265 mutation in these cases.The relationship between CD79B,MyD88 mutation and the clinicopathological features was analyzed.Results Immunophenotypically,15 cases were non germinal center B cell type.CD79B (Y196) mutation was detected in 4 cases (26.7％).For MyD88,L265 mutation was found in 7 cases (46.7％).CD79B and MyD88 mutations were found in 3 cases.The followup information was obtained in 8 patients.No association was found between CD79B,MyD88 mutation and outcome of patients.Conclusion Primary testicular diffuse large B cell lymphoma of non germinal center B cell type is a rare aggressive B cell lymphoma with poor prognosis and poor response to chemotherapy.CD79B,MyD88 gene mnutation was detected in Chinese patients with frequency of 26.7％ and 46.7％ respectively.It is possible for molecular targeted therapy of the primary testicular diffuse large B cell lymphoma on the basis of high frequency of CD79B and MyD88 gene mutation.

OBJECTIVETo investigate the application of double-step distraction osteogenesis in the reconstruction of mandibular segmental defects after tumor resection.

METHODSFrom January 2002 to December 2006, six cases of post-tumor unilateral mandibular segmental defects were reconstructed using distraction osteogenesis. The mandibular body was lengthened first, following by mandibular ramus distraction.

RESULTSNo infection or other complication was observed. The maximal distraction length reached 55 millimeter in the mandibular body, and 42 millimeter in the mandibular ramus. The average distraction length was 52 millimeter in the mandibular body, and 34 millimeter in the mandibular ramus. Both the aesthetic and functional result was excellent in all cases.

CONCLUSIONSDouble-step distraction osteogenesis is effective and easily performed in the reconstruction of unilateral mandibular segmental defects with less morbidities and complications. There is no need for donor site. However, the treatment period is relatively long with three staged operations.

Adolescent ; Adult ; Female ; Humans ; Male ; Mandibular Neoplasms ; surgery ; Middle Aged ; Osteogenesis, Distraction ; methods ; Reconstructive Surgical Procedures ; methods ; Young Adult

OBJECTIVETo study the effect of tumor suppressor gene PTEN on proliferation and cell cycle of hepatocellular carcinoma cell line HHCC.

METHODSFirstly, eukaryotic expression vectors of wild type and mutated type of PTEN gene were constructed, named as pEGFP-WT-PTEN and pEGFP-PTEN; G129R, respectively. Lipofectamine 2000 was used to transfect the constructed expression vectors into hepatocellular carcinoma cell line HHCC which was PTEN protein negative. G418 was used to select the cell clones constantly expressing PTEN protein. Flow cytometry was used to assay the cell cycle of HHCC transfected by above mentioned eukaryotic expression vectors and non-transfected cell line HHCC. Intrinsic 473-phosphorylated AKT representing the level of active AKT was assayed by Western blot. The non-transfected HHCC served as control.

RESULTSThe proliferation of HHCC constantly expressing PTEN protein was obviously inhibited compared with HHCC cells transfected with mutated PTEN gene and empty vectors, and non-transfected HHCC cells. The number of HHCC cells transfected with wild type PTEN gene at G(1) phase, G(2) phase and S phase was 70.8%, 6.8% and 22.4%, respectively. Compared with control group transfected with empty vector, the number of G(1) phase HHCC cells constantly expressing wild type-PTEN protein was significantly higher than that of control. The number of cells in G(2) and S phase was significantly lower than that of control. However, the number of cells in G(1) phase, G(2) phase and S phase of HHCC transfected with mutant PTEN was 63.2%, 10.1% and 26.7%, respectively. There was no significant difference compared with control group. Western blot result showed that the intrinsic level of 473-phosphorylated AKT of HHCC constantly expressing wild type PTEN protein was down-regulated, and that of HHCC transfected with mutated PTEN gene was equal to that of control.

CONCLUSIONWild type PTEN gene can inhibit the proliferation of hepatocellular carcinoma cells at G(1) phase. The mechanism is possibly related with intrinsic activity of AKT, which is down-regulated by wild type PTEN.

Carcinoma, Hepatocellular ; pathology ; Cell Division ; drug effects ; Cell Line, Tumor ; Genes, Tumor Suppressor ; Humans ; Liver Neoplasms ; pathology ; PTEN Phosphohydrolase ; genetics ; pharmacology

OBJECTIVETo investigate the effect of hydrogen sulfide-induced delayed preconditioning on glutathione S-transferase (GST) expression during myocardial ischemia-reperfusion in rats.

METHODSSprague-Dawley male rats were randomly divided into 4 groups (n= 10 in each): Group S (sham operation group), Group IR (ischemia/reperfusion group), Group H (IR+ NaHS 0.05 mg/kg iv, 24 h before ischemia) and Groups D receiving IR+NaHS 24 h before ischemia and 5-hydroxydecanoate (5-HD)15 min before ischemia. Animals in groups IR, H and D were subjected to ischemia by 30 min of coronary artery occlusion followed by 2 h of reperfusion. At the end of the reperfusion, myocardial infarct size (IS) was examined. Glutathione S-transferase (GST) was measured by Western blotting. The myocardial ultrastructures were observed under the electron microscopy.

RESULTSThe IS was significantly smaller in Group H than that in Group IR [(25.40 ± 3.54)% compared with (38.27 ± 5.64)%, P<0.05]. The GST expression in myocardium was significantly higher in Group H than that in Group IR. Microscopic examination showed less myocardial damage in Group H than in Group IR. The protective effects of delayed preconditioning by hydrogen sulfide was prevented by 5-HD pre-treatment.

CONCLUSIONThe hydrogen sulfide-induced delayed preconditioning attenuates myocardial IR injury possibly through up-regulating glutathione S-transferase expression in rats.

Animals ; Disease Models, Animal ; Glutathione Transferase ; metabolism ; Hydrogen Sulfide ; administration & dosage ; therapeutic use ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Reperfusion Injury ; enzymology ; pathology ; therapy ; Myocardium ; enzymology ; ultrastructure ; Rats ; Rats, Sprague-Dawley

Warfarin was the only oral anticoagulant drug , widely used in the prevention of thromboembolism events.The antithrombotic effect of warfarin has been very clear , but there are also some defects , such as that the treatment window is narrow and individual difference is very big.In recent years , the emergence of new targeted oral anticoagulant agents has provided a new choice for the anticoagulant drugs.The development of targeted oral anticoagulant drugs is reviewed , and the new challenge to warfarin is discussed.

OBJECTIVETo study the immune effect of recombinant adeno-associated virus (rAAV) combined with recombinant adenovirus (rAdV) vaccine in BALB/c mice.

METHODSThe codon-modified HIV-1 gp120 gene was inserted into plasmid of adeno-associated virus and adenovirus vector separately. Then the rAAV and rAdV vaccines were constructed. BALB/c mice were immunized with rAAV and rAdV vaccines in different administration scheme. The IgG antibody was detected by ELISA and CTL response was detected by intracellular cytokine stain assay.

RESULTSBoth rAAV and rAdV vaccine could express gp120 gene; the mice primed with rAAV at week 0, 2 and boosted with rAdV at week 5, 14 and 20 elicited the strongest gp120 specific CTL and IgG antibody response.

CONCLUSIONThe mice primed with rAAV and boosted with rAdV could elicit specific CTL response and IgG antibody.

Adenoviridae ; genetics ; Animals ; CD8-Positive T-Lymphocytes ; immunology ; Dependovirus ; genetics ; Female ; Genetic Vectors ; HIV Envelope Protein gp120 ; biosynthesis ; genetics ; immunology ; HIV-1 ; Immunoglobulin G ; blood ; Interferon-gamma ; blood ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombination, Genetic ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, DNA ; immunology ; metabolism

OBJECTIVETo assess the effect of surgical reconstruction of congenital aural atresia via the mastoid antrum approach and investigate method for preventing postoperative atresia of the reconstructed aural canal.

METHODSFrom 2000 to 2008, aural canal reconstruction and tympanoplasty was performed via the mastoid antrum approach. In 48 patients with congenital aural atresia (54 ears, including 45 ears of type II, 9 ears of type III). All the patients were followed-up for 18 months to assess the therapeutic effect.

RESULTSThe mastoid antrum was located uneventfully for all the 54 ears, all showing ossicular chain anomalies involving most frequently the malleus and the incus followed by the upper structures of the stapes. Facial nerve abnormalities were seen in 23 ears (42.6%). Hearing improvement to over 20 dB was achieved in 45 ears (83.3%) and to over 25 dB in 25 ears (46.2%) one year later.

CONCLUSIONThe mastoid antrum approach for surgical reconstruction of congenital aural atresia is safe and reliable. Maintenance of the width of the aural canal and prevention of lateral healing of the transplanted tympanic membrane are crucial in the treatment of congenital aural atresia.

Child ; Child, Preschool ; Ear Canal ; abnormalities ; surgery ; Ear, External ; abnormalities ; surgery ; Ear, Middle ; abnormalities ; surgery ; Female ; Humans ; Male ; Mastoid ; surgery ; Reconstructive Surgical Procedures ; methods ; Tympanoplasty

OBJECTIVETo study mutations of tumor suppressor gene PTEN in human hepatocellular carcinomas and its effects on the proliferation and apoptosis of hepatocellular carcinoma cell line HHCC.

METHODS(1) PCR-SSCP and sequence analysis were used to detect the mutations of the 5th and 8th exon of PTEN in 42 cases of human primary hepatocellular carcinoma. (2) Eukaryotic expression vectors of the wild-type (pEGFP-wt-PTEN) and the mutant type (pEGFP-PTEN, G129R) of PTEN were constructed. Lipofectamine 2000 mediated gene transfection was used to transfect hepatocellular carcinoma cell line HHCC, in which the PTEN protein is not expressed. Culture medium containing G418 was used to select stable transfectants. MTT colorimetry was used to analyze the proliferation ability of selected cell lines. Naive HHCC cells and HHCC cells transfected with empty vector (pEGFP-C1) served as controls. (3) TNF-alpha was used to induce apoptosis of selected cell clones.

RESULTS(1) Point mutation involving the 5th exon of PTEN was detected in 4 of 42 primary hepatocellular carcinomas. (2) Compared with the control groups, the proliferation of hepatocellular carcinoma cells was significantly inhibited by the transfection of wild-type PTEN gene, while the transfection with mutant PTEN construct did not significantly change the proliferation. (3) The apoptosis indices of cells transfected with the wild-type and the mutant PTEN genes were 13.8% and 8.1% respectively. Compared with the control, the apoptosis index of HHCC cell transfected by the wild type PTEN was significantly lower (P < 0.05). There were no significant differences between HHCC cells transfected with mutated PTEN gene and the control (P > 0.05). The expression of internal 473-phosphorylated Akt of HHCC was weak, but was enhanced when the cells treated with TNF-alpha. However, it was down regulated by the wild type PTEN.

CONCLUSIONS(1) First time report that PTEN mutations can be found in 9.5% human primary hepatocellular carcinomas. (2) The expression of the wild-type PTEN can suppress the proliferation of HHCC cells, and such suppression was lost when PTEN gene was mutated. (3) PTEN inhibition of the proliferation and the enhancement of apoptosis of hepatocellular carcinoma cells is likely related to a down-regulation of the TNF-alpha induced activation of protein kinase Akt pathway.

Apoptosis ; drug effects ; genetics ; physiology ; Base Sequence ; Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Mutational Analysis ; Flow Cytometry ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Microscopy, Fluorescence ; Molecular Sequence Data ; Mutation ; PTEN Phosphohydrolase ; genetics ; metabolism ; Transfection ; Tumor Necrosis Factor-alpha ; pharmacology