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

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

OBJECTIVETo prepare monoclonal antibodies (McAbs) against human programmed death-1 ligant 1(PD-L1) and identify its bioactivity.

METHODSWe immunized the BALB/c mice with Thioredoxin-(PD-L1) recombination protein which expressed by prokaryotic system. Prepare hybridoma cell by hybridoma technology and used enzyme-linked immunosorbent assay(ELISA) and Western-blotting assays to select positive hybridoma identify cell. Competition inhibition ELISA was carried out to identify the special bioactivity of antibody.

RESULTS4 hybridoma cell strains which could secrete anti-(PD-L1) antibodies stably were selected. The McAbs has good affinity with its receptor. Purify anti-(PD-L1) with title 1:32 000 was obtained after large quantity preparation. At the same time we obtained 1 cell stain which could secret special anti-Trx McAbs.

CONCLUSIONSWe obtained anti-(PD-L1) McAbs with good bioactivity successfully, which lay the foundation for further study.

Animals ; Antibodies, Monoclonal ; immunology ; Antibody Specificity ; Antigens, CD ; immunology ; B7-H1 Antigen ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hybridomas ; Mice ; Mice, Inbred BALB C

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

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.

OBJECTIVESTo discuss the methods and outcome of shorting proximal femoral and total hip arthroplasty for Crowe IV dysplastic hip of adults.

METHODSFrom July 2000 to February 2006, 13 cases of osteoarthritis secondary to severe development dysplastic hip were treated by total hip replacement and the shorting proximal femoral.

RESULTSThe duration of follow-up ranged from 4 months to 55 months. The average score increased from 36.9 to 84.1 points after the surgery according to Harris. All the patients could walk independently. Their paces were improved obviously and the function of their hips was satisfactory.

CONCLUSIONSThe treatment by total hip arthroplasty and the shorting of posterior femoral is effective and efficient for osteoarthritis secondary to Crowe IV development dysplastic hip in adults. The long-term followup is necessary for further study.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femur ; surgery ; Follow-Up Studies ; Hip Dislocation ; etiology ; surgery ; Humans ; Male ; Middle Aged ; Osteotomy ; methods ; Retrospective Studies ; Treatment Outcome

BACKGROUNDHuman urate anion exchanger (hURAT1) as a major urate transporter expressed on renal tubular epithelial cells regulates blood urate level by reabsorbing uric acid. Antibody is an important tool to study hURAT1. This study aimed, by genetic immunization, to produce mouse anti-hURAT1 polyclonal antibody with high throughput and high specificity and to detect the location of hURAT1 in human kidney.

METHODSHuman renal total RNA was isolated and the entire cDNA of hURAT1 was amplified by RT-PCR. The sequence of intracellular high antigenicity fragment (A280 to R349) was chosen by prediction software of protein antigenicity, and its cDNA was amplified from cDNA of hURAT1, and then cloned into pBQAP-TT vector to construct recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization. Mice were inoculated with this recombinant plasmid and two other adjuvant plasmids, pCMVi-GMCSF and pCMVi-Flt3L, which helped to enhance the antibody's generation. After four weeks, the mice were sacrificed to obtain the anti-hURAT1 antibody from serum. The antibody was identified by western blot analysis and immunohistochemistry. At the same time, rabbit anti-hURAT1 antibody was produced by protein immunization. The specificity and efficiency between the rabbit and mouse anti-hURAT1 antibody were compared by western blot analysis and immunohistochemistry.

RESULTSThe entire cDNA of hURAT1 and cDNA of its intracellular high immunogenic fragment were amplified successfully. Recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization was confirmed by restriction digestion and sequencing. Both the mouse anti-hURAT1 antibody and rabbit anti-hURAT1 antibody recognized 58 kD hURAT1 and 64 kD glycosylated hURAT1 protein bands in western blot. Immunohistochemically, hURAT1 was located at the brush border membrane of renal proximal tubular cells. In addition, the throughput and specificity of the mouse anti-hURAT1 antibody were higher than those of the rabbit anti-hURAT1 antibody.

CONCLUSIONGenetic immunization can generate anti-hURAT1 polyclonal antibody of high throughput and specificity.

Animals ; Antibodies ; analysis ; Blotting, Western ; Carrier Proteins ; analysis ; immunology ; Female ; Humans ; Immunization ; Immunohistochemistry ; Kidney ; chemistry ; Male ; Mice ; Organic Anion Transporters ; analysis ; immunology ; Organic Cation Transport Proteins ; Plasmids ; Rabbits

OBJECTIVETo investigate the related factors of portal vein tumor thrombosis (PVTT) in patients with HCC.

METHODSA total number of 234 patients with hepatocellular carcinoma (HCC) were included in this retrospective study. Uni-variate and multi-variate logistic regression analysis were employed to analyze the association between PVTT and 18 routine clinical parameters.

RESULTSAmong the 234 patients with HCC, 15% of patients (35/235) had PVTT. Univariate logistic regression analysis revealed significant association of age (P = 0.016), gamma glutamyl transferase (GGT, P = 0.003), number of segmental invasion (P = 0.007), microvascular invasion (P < 0.01), segment location of S2 (P = 0.001), S3 (P = 0.000), S4 (P = 0.004) and S6 (P = 0.016). Multivariate analysis shows potential significant predictors of PVTT in HCC were age (RR: 0.373; 95% CI: 0.146-0.954; P = 0.040), the tumor location of S3 (RR: 4.625; 95% CI: 1.916-11. 165;P = 0.001), GGT (RR: 4.091; 95% CI: 1.448-11.553; P = 0.008) and microvascular invasion (RR: 20.912; 95% CI: 4.745-92.172; P < 0.01).

CONCLUSIONSPVTT occurred more commonly in the younger (< 50 years old), and those with high level of GGT, segment location of S3 and microvascular invasion.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; pathology ; Child ; Embolism ; etiology ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; pathology ; Logistic Models ; Male ; Middle Aged ; Portal Vein ; Retrospective Studies ; Risk Factors