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

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

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.

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.

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.

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 Study the effect of Gross Saponins of Tribulus Terrestris on active potential and potassium current of ischemic cardiocytes in guinea pigs,and approach its initial anti-arrhythmia effect and mechanism.Methods The cardiocytes of adult guinea pigs were isolated and hypoxia models were set up,the effects of injected GSTT on the cellular membrane active potential and electricity flow of potassium channel were recorded with whole cell patchclamp system.Results Compared with model group,the active plateau period(50% and 90%) of myocytes in GSTT 30 mg?L-1 group prolonged(P

Objective To explore the role of plasminogen activator inhibitor-1(PAI-1)in development of progressive fibrosis via the inhibition of extracellular matrix degradation,and to reveal the contributive role of PAI-1 in muscular dystrophy(MD).Methods Expression and cellular localization of PAI-1 protein were examined in frozen muscle specimens obtained via biopsy from 5 patients with duchenne muscular dystrophy(DMD),3 patients with becker muscular dystrophy(BMD),9 patients with congenital muscular dystrophy(CMD) and 4 cases with normal muscle by immunohistochemistry,double immunofluorescence and Western-blot analysis.Results PAI-1 was positive only in vascular endothelial cells of normal muscle.Both immunohistochemistry and Western-blot analysis showed that PAI-1 expression distinctly increased in most dystrophic muscles of MD than that in normal muscles.Double immunolabeling revealed that PAI-1 strongly expressed in cytoplasm and nuclei of regenerating muscle fibers,macrophages,macrophage infiltrating necrotic fibers.Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were positive for PAI-1.Conclusions The functional consequence of overexpression of PAI-1 in dystrophic muscles is unknown but the elevated local expression of PAI-1 in diseased muscles of MD and their distinct distribution pattern provide evidence that PAI-1 participate in pathogenesis of MD.

Objective To preliminarily evaluate the efficacy and safety of chemotherapy on hemophagocytic lymphohistiocytosis(HLH) with hepatic dysfunction in children.Methods The children diagnosed as non-malignancy-associated HLH from Mar.2004 to Apr.2008 were selected,and the therapeutic effect was evaluated according to the HLH-04 protocol at the 8th week of chemotherapy,and the level of serum alanine aminotransferase(ALT),serum albumin(Alb) and plasma fibrinogen(Fib) were detected at pretherapy,2 weeks and 8 weeks of post-treatment.Results Altogether 60 HLH children complicated with hepatic dysfunction before chemotherapy,47 children had increased ALT,58 children had decreased Alb,and 38 children had decreased Fib.Forty-two cases(70%) were virus-associated HLH,1 case(1.7%) was fungi-associated HLH,and 17 cases(28.3%) had unknown origin.Among the 60 children,55 cases showed improvement in the 4 weeks of inductive treatment,15 cases gave up therapy,45 cases completed the 8 weeks of inductive treatment according to the protocol(among these children,42 cases had no active disease,3 cases had active disease),and these 45 children had obviously improved ALT,Alb and Fib at 2 weeks and 8 weeks of post-treatment,compared with pretherapy,the differences had statistical significance(Pa