Photoaffinity labeling is widely applied to demonstrate targets of small molecule ligands. In this paper, biotin photoaffinity labeled molecule with propargyl group 1 has been designed and synthesized, followed it's labeling of N2-acetyl-2'-O-propargyl guanosine 9 by "click chemistry". This technology presents delight development potential in labeling of second messenger cyclic nucleotide, antisense oligonucleotide or siRNA.
Biotin ; chemistry ; Click Chemistry ; Guanosine ; chemical synthesis ; chemistry ; Ligands ; Photoaffinity Labels

Objective To explore the ultrasound performance and related factors on the role of normal living rabbit's liver by laser ablation. Methods The rabbit's liver tissue were ablated by Echolaser integrated laser interventional ultrasound system, and the necrosis of the lesion and performance of pathology and anatomy were observed. Results The outline of the lesion was ellipse like. The two-dimensional US showed regular hyperecho area in the center, mild strong echo in the peripheral and mild attenuation backward. Contrast enhanced ultrasound (CEUS) showed a filling defect of contrast media in the ablated area. After dissection, the center of the lesion was slag-like carbon, the peripheral was necrosis area; HE staining showed: the center of the lesion was cavity like and dye-free,peripheral area was irregular red staining, the surrounding area was infiltrative inflammatory cells. Different power and time leaded to differences of the ablative effect and lesion size:the more power and time,the bigger of the ablative size. The ablative effect and lesion size was stable in 3 W 10 min and 5 W 6 min groups and caused the complete necrosis of the zone, there existed statistical differences among the two groups. Conclusions Laser ablation can cause fast, precise, effective and safe necrosis of the liver tissue, and the more power and time, the bigger of the ablative size.

Aged, 80 and over ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Male ; Prosthesis Failure

Adenoma ; diagnosis ; Child ; Diagnosis, Differential ; Humans ; Hypothyroidism ; pathology ; Magnetic Resonance Imaging ; Male ; Pituitary Gland ; pathology ; Pituitary Neoplasms ; diagnosis

This study was aimed to explore the anti-leukemic effect of scutellaria extract SBX in human leukemia cell lines and its mechanism. The leukemia cell lines, including HL-60, NB4, U937, K562 and Jurkat, were cultured in vitro and proliferative inhibition of these cell lines was detected by CellTiter-Glo Luminescent Cell Viability Assay in order to screen the most sensitive cell line. The effect of SBX on cell cycle was analyzed by flow cytometry and the protein expressions determined by Protein Pathway Array respectively. The results indicated that SBX (10 - 200 µmol/L, for 72 h) significantly inhibited the proliferation of different leukemia cell lines in a dose-dependent manner (r value was 0.86, 0.88, 0.95, 0.94, 0.96, respectively), the HL-60 was the most sensitive cell line. Flow cytometric analysis showed that SBX (50, 10 µmol/L, for 48 h) arrested HL-60 cells in the G(0)/G(1) phase. In addition, protein expression of p-PKC α/βII, p-p38, Cdc25B, XIAP of HL-60 cells increased, and p-AKT, p-SAPK/JNK, Notch4, Cdk4, Cdc2, cyclin E, Akt, Bcl-2, Bax, cdc42, TNF-α, p27, CaMKKa decreased after exposure to SBX (50 µmol/L, for 48 h). It is concluded that SBX can inhibit the proliferation of different leukemia cell lines, and HL-60 is a sensitive cell line. SBX significantly influences EGFR, Ras/Raf/MAPK and Notch signaling pathway, through which effects the expression of cell cycle-related proteins resulting in arrest of HL-60 cells in G(0)/G(1).
Cell Cycle ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Leukemia ; drug therapy ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Scutellaria ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism ; bcl-2-Associated X Protein ; metabolism

BACKGROUNDBlocking the 4-1BB/4-1BB ligand (4-1BBL) signal may modulate the secretion of Th1/Th2 cytokines and prolong the survival of the grafts, which play a key role in organ transplantation tolerance. The aim of this study was to investigate the role of blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody (mAB) in acute rejection of rat orthotopic liver transplantation.

METHODSThe orthotopic liver transplantation model was set up, while male Lewis rats were used as liver donors and Brown-Norway rats as recipients. The recipient rats were intravenously injected with anti 4-1BBL mAB or isotype control antibody. Groups were monitored for graft survival after transplantation. Plasma chemistry, including aspartate transaminase (AST), alanine aminotransferase (ALT), and bilirubin (BIL), was assayed. The concentrations of interleukin (IL)-2, IL-10 and interferon (IFN)-gamma in plasma were also measured by enzyme-linked immunosorbent assay. Allograft histology images were collected under light microscope and electron microscope.

RESULTSIsotype antibody treated recipients exhibited elevated plasma levels of liver injury markers including AST, ALT and BIL, progressive portal and venous inflammation and cellular infiltration of the liver allografts, and a mean graft survival time (MST) of 10.9 days. Administration of anti 4-1BBL mAB resulted in a decrease in plasma levels of liver injury markers and the concentrations of IL-2, IL-10 and IFN-gamma. The histological grade of rejection on day 7 decreased and MST (17.3 days) increased substantially.

CONCLUSIONSThese results demonstrate that attenuation of acute rejection follows the blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody and strongly suggest it is a promising strategy to prevent progression of graft rejection by suppressing T cell-mediated immunity.

4-1BB Ligand ; immunology ; Alanine Transaminase ; metabolism ; Animals ; Antibodies, Monoclonal ; pharmacology ; therapeutic use ; Aspartate Aminotransferases ; metabolism ; Bilirubin ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Graft Rejection ; immunology ; prevention & control ; Graft Survival ; drug effects ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Liver Transplantation ; adverse effects ; Male ; Rats ; Rats, Inbred Lew

OBJECTIVETo evaluate the endourethral surgery for the complicated urethra stenosis and urethratresia.

METHODSThe endourethral surgery, such as internal urethrotomy transurethral scar electrosectomy or transurethral scar plasmakinetic bipolar electrocautery (PKR) or transurethral laser cicatrectomy, were carried out in 46 cases suffering from the complicated urethra stenosis and urethratresia.

RESULTSThe curative rate in this series being achieved by once and twice or three times'operation were 80.43% (39/46) and 13.04% (6/46) respectively. Three cases of treatment failure were caused by long-segment stricture and urethratresia or severe malposition of the urethral proximal and distal to a narrow-caliber area or post-operation infection. Thirty-nine cases have been followed up for 6 to 84 months. Satisfactory voiding has been achieved in all patients.

CONCLUSIONEndoscopic surgery was believed to be a safe and efficient therapeutic choice for the complicated urethra stenosis and urethratresia. The success of the treatment depends on understanding the length of the stricture before operation, resecting completely the scar tissue with electric or PKR or laser technique during the process, preventing infection and managing appropriately the urethral catheterization after operation.

Adolescent ; Adult ; Aged ; Endoscopy ; Follow-Up Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Retrospective Studies ; Urethra ; abnormalities ; surgery ; Urethral Obstruction ; surgery ; Urethral Stricture ; surgery ; Urogenital Surgical Procedures ; methods

Angiogenic gene therapy and cell-based therapy for peripheral arterial disease(PAD) have been studied intensively currently. This study aimed to investigate whether combining mesenchymal stem cells(MSCs) transplantation with ex vivo human hepatocyte growth factor(HGF) gene transfer was more therapeutically efficient than the MSCs therapy alone in a rat model of hindlimb ischemia. One week after establishing hindlimb ischemia models, Sprague-Dawley(SD) rats were randomized to receive HGF gene-modified MSCs transplantation(HGF-MSC group), untreated MSCs transplantation (MSC group), or PBS injection(PBS group), respectively. Three weeks after injection, angiogenesis was significantly induced by both MSCs and HGF-MSCs transplantation, and capillary density was the highest in the HGF-MSC group. The number of transplanted cell-derived endothelial cells was greater in HGF-MSC group than in MSC group after one week treatment. The expression of angiogenic cytokines such as HGF and VEGF in local ischemic muscles was more abundant in HGF-MSC group than in the other two groups. In vitro, the conditioned media obtained from HGF-MSCs cultures exerted proproliferative and promigratory effects on endothelial cells. It is concluded that HGF gene-modified MSCs transplantation therapy may induce more potent angiogenesis than the MSCs therapy alone. Engraftment of MSCs combined with angiogenic gene delivery may be a promising therapeutic strategy for the treatment of severe PAD.
Animals ; Bone Marrow ; metabolism ; pathology ; Bone Marrow Transplantation ; Cells, Cultured ; Hepatocyte Growth Factor ; genetics ; Hindlimb ; pathology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Neovascularization, Physiologic ; genetics ; Rats

OBJECTIVETo investigate the distribution of ER isoforms in endometriosis and eutopic endometrium.

METHODSTissue samples of patients with ovarian endometriosis, treated in People's Liberation Army General Hospital from January 2004 to December 2006, were retrieved. A total of 60 cases of ovarian endometriotic cysts with their corresponding eutopic endometrium (30 cases of proliferation phase and 30 of secretary phase eutopic endometrium) and 30 cases of normal endometrium (15 proliferative and 15 secretary phase endometrial samples respectively) were included. Expressions of ERalpha and ERbeta were analyzed using immunohistochemistry and the expression ratio was statistically analyzed by using SPSS 12.0 software.

RESULTSExpressions of both ERalpha and ERbeta in epithelial cells were positively correlated with that of the stromal cells. The expression of ERalpha in eutopic endometrium (73.3% in epithelium and 76.7% in stroma) was significantly higher than that in ovarian endometriotic cysts (43.3% in epithelium and 46.7% in stroma), or normal control (56.7% in epithelium and 50.0% in stroma, respectively, each P < 0.05. However, the expression of ERbeta (90.0% in epithelium and 76.7% in stroma) was higher in ovarian endometriotic cysts than that in the eutopic endometrium (68.0% in epithelium and 63.3% in stroma respectively, P < 0.05), and ERbeta expression in eutopic endometrium was higher than that in the normal control endometrium (36.7% in epithelium and 26.7% in stroma, respectively, P < 0.05). The expressions of both ERalpha and ERbeta changed periodically in eutopic and normal endometrium, whereas ERalpha and ERbeta level were less variable in the ectopic endometrium. The expression of ERbeta was statistically higher than that of ERalpha (P < 0.05) in ectopic endometrium, whereas no significant difference was seen between the two isoforms in the eutopic or normal endometrium.

CONCLUSIONSBoth ERalpha and ERbeta have higher expression levels in eutopic endometrium of patients with ovarian endometriotic cysts. ERbeta is predominantly expressed in endometriotic cysts, where the expression of ERalpha is limited. The different distribution of ERalpha and ERbeta may play an important role in the development of ovarian endometriosis.

Adult ; Choristoma ; pathology ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; pathology ; Epithelium ; Estrogen Receptor alpha ; analysis ; Estrogen Receptor beta ; analysis ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Protein Isoforms ; analysis ; Receptors, Estrogen ; analysis ; Stromal Cells ; metabolism

OBJECTIVETo estimate the efficacy of Gamma knife radiosurgery (GKR) especially as a primary surgical treatment for hypersecreting pituitary adenoma.

METHODSOne hundred and twenty cases with hypersecreting pituitary adenoma had been treated by Gamma knife radiosurgery. The clinical date had been analysed retrospectively. The tumor margin was covered by an isodose ranging from 45% to 70%. The margin dose was 15 to 32 Gy (mean 28.5 Gy) and the maximum dose varied from 35 to 70 Gy (mean 45.5 Gy). The total number of isocenter was 165 (mean 1-3).

RESULTSOne hundred and eleven cases had been followed-up by hormone level, and 104 cases had been followed-up by image of MRI. The mean follow-up duration was 12-72 months (mean 36 months). The control rate of hormone level was 48.6%, the control rate of tumor growth was 96.2%, the incidence of hypopituitarism was in 2.7% and the incidence of tumor apoplexy was in 0.9% in followed-up cases.

CONCLUSIONSAs a primary surgical treatment for hypersecreting pituitary adenoma, GKR can be effective and safe in controlling tumor growth and inducing hormonal normalization.

Adenoma ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pituitary Neoplasms ; pathology ; surgery ; Radiosurgery ; methods ; Retrospective Studies ; Treatment Outcome