OBJECTIVETo investigate the expression of survivin mRNA in hematological malignancy cells and its correlation with HHT-induced cell apoptosis.

METHODSHematological malignancy cell lines MUTZ-1, K562, Jurkat, RPMI and HL60 were treated with HHT in vitro. Cell apoptosis was observed by flow cytometry (FCM) and the expression of surviving and XIAP mRNA was evaluated with semi-quantitative RT-PCR.

RESULTThe expression of survivin mRNA on cell lines was negatively correlated to HHT-induced cell apoptotic rate (r=-0.980, P=0.003). There were no significant differences in XIAP expression among these 5 cell lines.

CONCLUSIONSurvivin could be used as a new marker for drug-sensitivity and a new target for treatment of hematological malignancies.

Apoptosis ; drug effects ; HL-60 Cells ; Harringtonines ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; K562 Cells ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Myelodysplastic Syndromes ; metabolism ; pathology ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured

BACKGROUND: Cytochrome C oxidase(CCO) is the terminal enzyme in respiration chain of mitochondrion, and it plays a key role in aerobic metabolism and energy production during the process of oxidative phosphorylation. Recently, it is found that energy production of mitochondrion is closely related to the cellular apoptosis, and the changes of CCO activity is closely related to the neuronal impairment after cerebral ischemia and anoxia.OBJECTIVE: To investigate the protective mechanisn of compound pinggan xifeng decoction on the neuronal impairment in cerebral hemorrhage (CH) according to the mitochondrial energy metabolism and cellular apoptosis in neurons.DESIGN: A randomized and controlled trial based on experimental animals.SETTING: Institute of Integrated Traditional Chinese Medicine and Western Medicine, Xiangya Hospital, and Center of Telemedicine.MATERIALS: The experiment was completed in the animal laboratory(key laboratory of province) of Institute of Combination of Traditional Chinese Medicine and Western Medicine from November 2 to 9 in 2003. A total of 80 healthy male SD rats were selected from Experimental Animal Center of Xiangya School of Medicine, Public Health Ministry.METHODS: CH rat models were induced with collagenase Ⅶ, CCO activity was assayed with histochemistry combined with semi-quantification of gray scale, and the cellular apoptosis was evaluated with Tunel method.MAIN OUTCOME MEASURES: CCO activity of CH rats in lateral hippocampal CA1;lateral cellular apoptosis of CH rats.RESULTS: After 12-hour model establishment, CCO activity in CH group was decreased dramatically compared with that in sham operation group (P＜ 0.01), which was 52.12 ±3.75 and 26.98 ±6.32 respectively in lateral hippocampal CA1. And the cellular apoptosis in CH group was increased notably compared with that in sham operation group(P ＜ 0.01),which was(13.56 ± 1.72)/sight and(4. 32 ± 1.04)/sight respectively.Then the two had deteriorated afterwards. After the treatment with pinggan xifeng decoction, CCO activity can be maintained, and the cellular apoptosis was reduced.CONCLUSION: Neuronal injury was closely related to the decrease of CCO activity and the cellular apoptosis in CH. Pinggan xifeng decoction could maintain CO activity of mitochondrion, improve the cellular aerobic netabolism, and reduce the cellular apoptosis, which might be one of the protective mechanisms for secondary neuronal injury in CH.

Objective To investigate the effects of different degrees of hemodilution with the artificial plasma substitutes most commonly used in clinical practice on coagulation in vitro. Methods Ten male ASA physical status I volunteers aged (28.8?1.6) yr and weighing (66?8) kg were enrolled. Venous blood samples obtained from each volunteer were diluted with 10% HES (200 000/0.4-0.55), 4 % succinylated gelatin (gelofusine GEL), 3.5 % polygeline (Haemaccel HAE) and lactated Ringer's solution (RL) by 33% [blood: diluent(B:D) =2:1], 50% (B:D=1:1) and 66% (B:D=1:2). Coagulation of the undiluted blood (control) and diluted blood was measured with sonoclot coagulation and platelet function analyzer (SCT) and by routine coagulation tests. The parameters measured included activated clotting time (ACT), clot rate, platelet function (PF), Hct, platelet count (Pit), plasma fibrinogen concentration (Fig) and activated partial thromboplastin time (APTT) . Results (1) At 33 % dilution ACT was significantly shortened in all the four groups as compared with control value; at 50 % dilution ACT was significantly shorter than the control in RL, HES and HAE groups; at 66 % dilution ACT was significantly prolonged in HES group. (2) Clot rate was significantly decreased at 33 % dilution only in HES group but was significantly decreased in all the four groups at 50 % and 66 % dilution. (3) PF decreased significantly only in GEL group at 33 % dilution but was significantly decreased in GEL and HES groups at 50 % and 66 % dilution. (4) With increasing dilution Hct, Pit and Fig gradually decreased and APTT was prolonged. Conclusion Coagulation changes are closely related to the degrees of dilution. At 33 % dilution, the three artificial plasma substitutes tested do not significantly affect hemostasis. At 50 % and 66 % dilution coagulation is badly impaired, but there are differences among the substitutes used for dilution. HAE impairs oagulation least as it contains higher calcium

Aim To study the effect of Baoxinbao film on endothelin(ET) and nitric oxide(NO) secretion in patients with stable angina pectoris(SAP).Methods 76 patients with SAP were randomly divided into two groups, with 40 cases in the baoxinbao group plastered with baoxinbao film and 36 cases in the isosorbide dinitrate group receiving isosorbide dinitrate. The levels of plasma ET and NO before and after treatment were observed. Results The concentrations of plasma ET were increased and plasma NO reduced significantly in the SAP patients respectively, as compared with those in the control group(all P

OBJECTIVETo investigate the expression profile of interleuki-1β (IL-1β) in rat myocardium at different time points during hypoxia/reoxygenation(H/R)transition.

METHODSThe isolated Langendorff perfused rat heart model was established.Forty SD rats were randomly divided into sham group (A group) and hypoxia/reoxygenation group (H/R group). The H/R group rats were subdivided into H/R 0.5 h group(B group), H/R 1 h group(C group), H/R 2 h group(D group)according to reoxygenation time. The left ventricular development pressure(LVDP), maximal rates of increase/decrease of the left ventricular pressure(±dp/dtmax) were continuously recorded. The concentration of interleukin-1β(IL-lβ) and creatine kinase-MB (CK-MB) in myocardium was measured by ELISA. The mRNA expression of IL-lβ in myocardium was determined by RT-PCR. Microstructure of myocardium was observed under light microscopy.

RESULTSThe value of LVDP and ±dp/dtmax in hypoxia/reoxygenation group rat were significantly lower than that in sham group(P < 0.05). The expression of IL-lβ and CK-MB at protein level and the expression of IL-1β at mRNA level in hypoxia /reoxygenation group were higher than that in sham group(P < 0. 05). There were significant differences of the above parameters among H/R 0.5 h, 1 h, 2 h group(P <0.05). The concentration of IL-1β and CK-MB, the mRNA expression of IL-1β were higher in H/R 2 h group than that of other groups(P < 0.05).

CONCLUSIONThe high expression of IL-Iβ in myocardium after myocardial hypoxia /reoxygenation in rats might lead to. ischemia/reperfusion injury.

Animals ; Creatine Kinase, MB Form ; metabolism ; Disease Models, Animal ; Hypoxia ; metabolism ; pathology ; Interleukin-1beta ; metabolism ; Myocardial Ischemia ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the expression of apoptotic protein inhibitors, survivin and XIAP, in patients with myelodysplastic syndromes (MDS) and in the cell line MUTZ-1, as well as to explore the possible mechanisms of homoharringtonine (HHT) in the treatment of MDS.

METHODSBone marrow samples from 47 patients with de novo MDS at diagnosis were examined and bone marrow samples from 15 normal donors were used as control. A MDS-RAEB cell line MUTZ-1 was used as in vitro model. Detection of apoptotic cells and cell cycle analysis were performed with flow cytometry (FACS). The expression of apoptotic protein inhibitor survivin and XIAP in the MDS cells were detected by RT-PCR technique. MUTZ-1 were treated with antisense oligodeoxynucleotide (AS-ODNs) of survivin and or HHT, the effects were evaluated by cell viability and cell apoptosis.

RESULTSSurvivin mRNA positive rate in MDS were significantly higher than that in normal controls (38.3% and 0, respectively, P < 0.01), and the positive rate in high risk group (RAEB, RAEBT and CMML) was significantly higher than that in RA/RAS group (53.6% and 16.7%, respectively, P < 0.05). XIAP was expressed in all untreated MDS and healthy controls. XIAP mRNA expression in high risk group was significantly higher than that in RA/RAS subtypes and healthy controls (1.55 +/- 0.34, 0.74 +/- 0.24, and 1.01 +/- 0.28, respectively, P < 0.01). However, XIAP mRNA expression was significantly lower in RA/RAS subtypes than in healthy control (0.74 +/- 0.24 and 1.01 +/- 0.28, P < 0.054). Apoptosis peak detected by FACS analysis and positive Annexin V FITC staining on cell membrane indicated that HHT could induce MUTZ-1 cell undergoing apoptosis in dose- and time-dependent manners. Treatment of MUTZ-1 cells with HHT revealed that HHT could significantly down-regulate survivinexpression but had no significant effect on XIAP expression in the cells. AS-ODNs of survivin could inhibit MUTZ-1 cells growth, induce them to apoptosis and sensitize them to HHT.

CONCLUSIONThe expression levels of survivin; Institute of Hematology, Oncology and Tumor Immunology, Robert Roessle Clinic, Humboldt University, Berlin, Germany (Wolf Dieter Ludwig, Christian Wuchter) and XIAP vary in different subtypes of MDS patients, suggesting that the proteins may play an important role in the pathogenesis of the disease. Down-regulation of survivin in MUTZ-1 cells may be one of the mechanisms that HHT induces apoptosis of MDS cells.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Harringtonines ; therapeutic use ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; physiology ; Myelodysplastic Syndromes ; drug therapy ; pathology ; Neoplasm Proteins ; Oligonucleotides, Antisense ; pharmacology ; Proteins ; genetics ; physiology ; RNA, Messenger ; analysis ; X-Linked Inhibitor of Apoptosis Protein

OBJECTIVETo investigate the TLR4 signaling in multiple myeloma cell proliferation and apoptosis.

METHODSTLR4 gene transcription and protein expression were detected by RT-PCR and PE flow cytometry staining; the cells proliferation were detected by MTT assay; doxorubicin-induced apoptosis of cells was detected by AnnexinV-PI double staining flow cytometry.

RESULTTLR4 gene transcription and protein expression was detected in the myeloma cell lines. Under the lipopolysaccharides (LPS) stimulation, MM1-s cells TLR4 positive proliferation significantly increased (P<0.05), but not found in U266 cells TLR4 negative; MM1-s cells showed the resistance to doxorubisin-induced apoptosis, but LPS did not protect doxorubicin-induced U266 cell apoptosis.

CONCLUSIONTLR4 signaling may play an important role both in multiple myeloma proliferation and survival.

Apoptosis ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Multiple Myeloma ; immunology ; metabolism ; pathology ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo study the expression of carbonic anhydrase (CA) IX and its significance in molecular subtyping of breast carcinomas. METHODL MaxVision immunohistochemical staining was used to examine the expression of ER, PR, HER2, CK5/6, EGFR, and CA IX in 117 cases of breast invasive ductal carcinomas.

RESULTSThe patients' age ranged from 25 to 71 years (mean 49.6 years). All the 117 cases were subclassified into five subtypes, with 66 (56.4%) luminal A, 6(5.1%) luminal B, 10 (8.6%) HER2 positive, 20 (17.1%) basal-like, and 15 (12.8%) unclassified tumors. The expression of CA IX in luminal A and basal-like breast cancers was 13.6% (9/66) and 8/20, respectively, with a significant difference (P < 0.05). Among the luminal A cancers, the expression of CA IX in tumors > 2 cm (7/27, 25.9%) was significantly (P < 0.05) higher than that of tumors ≤ 2 cm (2/39, 5.1%). The expression of CA IX in grade 3 invasive ductal carcinoma (18/50, 36.0%) was significantly higher than that in grade 1 (2/21, 9.5%) and 2 (7/46, 15.2%) tumors (both P = 0.006). In CA IX-negative of invasive ductal carcinoma, the expression of ER and PR was 61.1% (55/90) and 55.6% (50/90), respectively; whereas in CA IX-positive cancers, the expression of ER and PR was 37.0% (10/27) and 29.6% (8/27), respectively. The expression of hormone receptors in CA IX-negative tumors was significantly higher than that in CA IX-positive tumors (for both ER and PR, P < 0.05).

CONCLUSIONSThe expression of CA IX correlates not only with molecular subtypes of breast cancer, but also with the grading, hormone receptors and diameter of mammary invasive ductal carcinoma. CA IX is a relative independent marker of poor prognosis in breast cancer.

Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; classification ; metabolism ; pathology ; Carbonic Anhydrase IX ; Carbonic Anhydrases ; metabolism ; Carcinoma, Ductal, Breast ; classification ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Neoplasm Grading ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Tumor Burden

OBJECTIVETo evaluate the inhibitory effect of small interfering RNA (siRNA) targeting HBV C gene region on hepatitis B virus (HBV) in vivo.

METHODSAn animal model of HBV infection was developed hydrodynamically, and pcDNA3.1-HBV and siRNA were together injected into the tail vein of the BALB/c mice. HBsAg was analyzed by time-resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV C-mRNA was detected by semi-quantitative RT-PCR, and viral specific proteins (HBsAg and HBcAg) in the mice livers were assayed using immunohistochemical staining.

RESULTSIn the mice, the siRNA effectively inhibited HBV replication and expression compared with the controls. The inhibitive effect of siRNA on HBV lasted at least 3 days.

CONCLUSIONThese results demonstrate that RNAi can substantially inhibit HBV replication and expression in vivo.

Animals ; Female ; Hepatitis B ; therapy ; Hepatitis B virus ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; physiology ; RNA-Induced Silencing Complex ; Random Allocation ; Virus Replication ; genetics

OBJECTIVETo explore the multi-differentiated capability of human periodontal ligament cell population (hPDLP), and provide a theoretical basis for the periodontal regeneration by tissue engineering technique.

METHODShPDLP was cultured from periodontium of human tooth by the outgrowth method. STRO-1 and CD 146 expression were investigated by flow cytometry. hPDLP was induced to odontogenic/osteogenic-like and adipogenic-like cell. The multilineage differentiation capacities of hPDLP were evaluated by alizarin red stain, oil red O stain, anti-CD146 and STRO-1 immunocytochemistry, and reverse-transcriptase polymerase chain reaction analysis.

RESULTShPDLP was isolated from human periodontium and most of the cells retained their fibroblastic spindle shape. hPDLP can be induced into osteoblast-like cells and adipocyte-like cells, and calcium deposition and lipid droplets were detected perspectively. And the eighth generation of hPDLP had weaker potential into adipocyte-like cells than the first passage, however, there was no difference to the aspect of calcification ability between the two passages.

CONCLUSIONhPDLP cultured in vitro can differentiate into adipocytes and osteoblasts, and the first to third passage cells may have the predominance of differentiation potential.

Adipocytes ; Cell Differentiation ; Cells, Cultured ; Fibroblasts ; Flow Cytometry ; Humans ; In Vitro Techniques ; Odontogenesis ; Osteoblasts ; Periodontal Ligament ; Regeneration ; Tissue Engineering