Aim To observe the protection effect of Tribulus terrestris saponin monomer B (TTSMB) on cadiocytes impaired by hypoxia-reoxygenation (H/R).Methods Cadiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, H/R group,GSTT 100 mg·L~(-1) group and TTSMB 10,1,0.1 nmol·L~(-1) group.Morphocytology change of cadiocytes was observed after the treatment.Cadiocyte survival rate was detected with MTT colorimetric method.Levels of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), malondialdehyde (MDA), and activity of superoxide dismutase (SOD) were determined. Apoptosis rate was detected with flow cytometry.Expression of caspase-3 was examined with western blot.Results Compared with the control group, the survival cell number in H/R group decreased obviously, content of CK,LDH, AST, MAD increased, and activity of SOD decreased (P<0.01).Compared with the model group, the survival cell population increased in TTSMB 10,1,0.1 nmol·L~(-1) group (P<0.05 and P<0.01).Contents of CK, LDH, AST, MAD decreased, whereas the activity of SOD increased (P<0.05 and P<0.01).Apoptosis rate and expression of caspase-3 also reduced in TTSMB 10,1,0.1 nmol·L~(-1) group.Conclusion TTSMB has significant cadiocytes protective effect against H/R and inhibits cadiocyte apoptosis,and the mechanism depends on the effect against oxygen free radical.

Antibodies, Monoclonal ; analysis ; Biopsy, Needle ; Breast Neoplasms ; chemistry ; pathology ; Female ; Humans ; In Situ Hybridization ; methods ; In Situ Hybridization, Fluorescence ; Practice Guidelines as Topic ; Quality Control ; Receptor, ErbB-2 ; analysis ; immunology ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis ; Societies, Medical ; Specimen Handling ; United States

Objective To optimize the synthesis process of iloperidone 1 for industrial production.Methods Piperi-dine-4-carboxylic acid, acetic acid and acetic anhydride were used as starting materials to generate 2 via amidation, Friedel-Crafts acylation, hydration, oximation, intramolecular cyclization and salification.Compound 2 was treated with 4-(3-chloropropoxy)-3-methoxyphenylethanone through condensation to afford iloperidone.The target compound 1 was prepared with an overall yield of 34.9%.Results and Conclusion Such factors as the ratio of the raw materials, the catalyst, reactive time were investigated and optimized to make the reaction conditions mild and easy to control with less side effects.The structure has been confirmed by IR, 1 H-NMR and MS.

Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity. CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Nobiletin potently activated CFTR chloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTR(inh)-172 could completely reverse the effect. Preliminary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may be a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis.

Objective:To evaluate angioimmunoblastic T-cell lymphoma(AITL) completely, we gave in-depth investigation of histopathological features, specific immunochemical markers, antigen receptor gene rearrangements and in situ hybridization for Epstein-Barr virus (EBV). Methods: 15 cases of typical AITL displayed effacement of the normal lymph node architecture partially or completely, abundance of arborizing high endothelial vessels, infiltration of polymorphic cells and hyperplastic atypical T lymphocytes with or without clear cytoplasm. Clinical characteristics, histological manifestations, and immunohistochemical staining for CD3, CD20, CD4,CD21, CXCL13, CD10, and BCL6 were analyzed. Polymerase chain reaction for immunoglobulin heavy chain (IgH) and T cell receptor ? (TCR?) rearrangements and in situ hybridization for Epstein-Barr virus encoded RNA (EBER-1) were performed.Results: Histologically, we found eight cases with regressed lymphoid follicles, six with absence of follicles and one with hyperplastic follicles with interfollicular lesions. We also found eight cases displaying aggregation of clear cells, four infiltration of large lymphoid cells, five abundant epithelioid histiocytes. CD20 staining showed hyperplasia of large B cells in four cases. CD21 expression exihibited extrafollicular expansion of follicular dendritic cell meshworks in 11 cases (73.3%), partially with a tendency of perivascular distribution. Positive rate for CXCL13 and CD10 are 73.3% and 6.7% respectively. Monoclonal rearrangements of TCR? were detected in 6/15 (40%) of cases, IgH rearrangements in 7/15 (46.7%), of which five were monoclonal, while two oligoclonal. 8 out of 15 cases (53.3%) contained EBV-positive cells. Among the four cases with large B cell proliferation, three were EBV-positive. Conclusion: AITL display great complexity and diversity clinicopathologically. Only when we recognize such diversity, can we reasonably apply and properly evaluate immunochemical markers and molecular techniques, and thus give a correct diagnosis.

Objective To analyze the pathogenesis,therapy and outcome of pediatric cases with coagulation disorders (CD).Methods All these 137 patients were diagnosed as CD with the methods of hemoglutination five items and/or disseminated intravascular coagulation indexes.Then activity of specific coagulation factors,morphology of bone marrow,hepatorenal function and some other relative tests were performed to find out the cause of CD or the primary disease.Results Forty-three cases were diagnosed as genetic CD with 29 as hemophilia A,4 as hemophilia B and 10 as Von Willebrand disease;while the other 94 patients as acquired CD with 15 as vitamin K-dependence coagulation factor deficiency,22 as hepatic dysfunction,30 as disseminated intravascular coagulation and 1 as thrombotic thrombocytopenic purpura.Genetic CD was treated with replacement therapy to reduce the complication.There was 1 case in this group died of intracranial hemorrhage.Acquired CD was treated with short-term,specific and necessary replacement therapy on the basis of reasonable treatment of primary diseases.Eleven cases died finally in this cohort with 7 cases as liver failure and the other 4 cases as terminal leukemia or lymphoma.Conclusion Pediatric patients with CD were caused by genetic or acquired diseases.In clinic the reason of CD was mainly acquired.The treatment of genetic CD is the replacement of specific coagulation factor for life-long term.The outcome dependes on the lack of degree.While the therapy for acquired CD aims at the primary disease.The principle of blood transfusion is short-term and the outcome dependes on the therapic effects of primary diseases.

BACKGROUND: Multipotency of hepatic stem cells is of important value in liver transplantation. Stem cells have been successfully identified and isolated from the animal livers. However, reports on whether stem cells exist in human hepatic tissue and how to isolate and identify them ars few.OBJECTIVE: This study was in attempt to isolate hepatic stem cells from human para-cancerous tissues of hepatoma and in vitro culture them, also to identify the stem cell surface marker, in order to find a new source of heptatic stem cells.DESIGN: Cell observation experiment.SETTING: Department of Common Surgery, First Hospital, Jilin University; Department of Common Surgery,Dongfeng Hospital of Traditional Chinese Medicine.PARTICIPANTS: Samples were harvested from 10 patients with hepatoma admitted to Department of hepatobiliary surgery, First Clinical College, Jilin University between October 2005 and June 2006, with age of 45 to 58 years.Hepatic tissue 2 cm away from cancer nest was cut when patients underwent hepatectomy, and it was pathologically confirmed as carcinoma-free tissue. Written informed consents were obtained from each patient. DMEM/F12 dry powder used for cell culture was provided by Hyclone Company, USA. Fresh fetal bovine serum was prepared by Lianxing Biotech Co.,Ltd, Tianjin. Various cell growth factors were the products of Cytolay Company, USA.METHODS: Para-cancerous tissues of hepatoma was cut into pieces, rinsed with Hank's solution and digested with type Ⅳ collagenase. Then the isolated cells were re-suspended in the DMEM/F12 medium supplemented with 0.1 volume fraction of fetal bovine serum, and hepatocyte growth factors, epidermal growth factors and α- fibroblast growth factors of 25 μg/L each were added in the above medium. When the cultured cells covered 2/3 of bottom,they were digested with trypsinase for passage and inoculated at 2×107 L-1. When cells propagated to the 3rd and 4th generations, 2.60×109 L-1 cell suspension prepared with trypsinase was added, and subsequently, anti-human C-kit antibody, immunomagnetic beads and Buffer solution were added in order. C-kit+ cells were preliminarily isolated by immunomagnetic bead separation. Haematoxylin-eosin staining and immunofluorescent histochemical double-staining were used for detecting the hepatic stem cells in para-cancerous tissues.MAIN OUTCOME MEASURES: ① Observation of cell morphology. ② Identification of hepatic stem cells from para-cancerous tissues. ③ Identification of C-kit+ cells by immunofluorescent histochemical double-staining.RESULTS :① After primarily cultured for 2 weeks, the adherent cells grew in colony. After one half of culture medium was renewed, mature hepatocytes were gradually broken and disappeared. Small round cells propagated, and most of them were located in the center and arranged in cluster. Most cells were found with one big nucleus in each, less cytoplasm and clear cell boundary. When cells propagated to the 1st and 2nd generations, they still grew in colony, but fast. Each C-kit+ cell isolated by immunomagnetic bead separation presented a spherical cell body with a very big nucleus and less cytoplasm. After in vitro cultured for 1 week, it presented broken pieces and apoptotic symptoms.② After para-cancerous tissue was stained by haematoxylin-eosin, atypically proliferated biliary tracts with small round cells could be seen in the portal area. After para-cancerous tissue was stained by immunofluorescent histochemical double-staining, small round cells in the biliary tracts proliferated in the portal arsa co-expressed red fluorescence AFP and green fluorescence cytokeratin (CK) 19 with yellow superposition arsa. ③ After C-kit+ cells were stained by fluorescence immunocytochemisty, cytoplasm expressed alpha-fetoprotein (AFP) red granules and CK19 green granules. The superposition area of both presented yellow fluorescence of AFP+/CK19+-positive cells.CONCLUSION: Hepatic stem cells exist in human para-cancerous tissues of hepatoma. Therefore, expressions of C-kit+/AFP+/CK19+, the surface markers of hepatic stem cells, can be used for identifying and isolating hepatic stem cells. Small round cells obtained by in vitro isolation and culture, i.e. hepatic oval cells possess bipotential differentiation of hepatocyte and hepatobiliary epithelial cells.

This study is to observe the protection of gross saponins of Tribulus terrestris (GSTT) on cardiocytes impaired by adriamycin (ADR) and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model (ADR 2 mg x L(-1)) group, and GSTT (100, 30, and 10 mg x L(-1)) groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced (P < 0.05, P < 0.01, P < 0.001) by ADR. Numbers of survival cells were increased (P < 0.05, P < 0.001), contents of CK, LDH, AST, MDA and NO were decreased, and activity of SOD was increased (P < 0.05, P < 0.01, P < 0.001) by GSTT (100 and 30 mg x L(-1)). Apoptosis of cardiocytes and concentration of caspase-3 can be reduced by GSTT (100 and 30 mg x L(-1)). GSTT can protect cardiocytes impaired by ADR, which are possible involved with its effect of resisting oxygen free radical.

Evolution, Molecular ; Methicillin Resistance ; Staphylococcus aureus ; classification ; drug effects ; genetics ; isolation & purification

Stroke is one of the severe perioperative complications.The incidence of perioperative stroke has great difference because of the surgical site,surgical complexity,and different basic diseases of patients.The induced anesthesia and the intraoperative management also have an important influence on the onset of stroke.The mortality of perioperative stroke is as high as 26%.Neturologists must guide the assessment of the potential risks for perioperative stroke,and carefully balance the risk-benefit ratio when making each decision.This article reviews the advances in research on the risk factors for perioperative stroke,prevention,and treatment.