Objective: To detect the abnormal expression of WWOX (WW domain containing oxidoreduc-tase) exons 6-8 at mRNA level in human benign and malignant pleural effusion and to investigate the role of loss of WWOX exons 6-8 in the development of malignant pleural effusion. Methods; Deletion of WWOX ex-ons 6-8 mRNA transcript was analyzed by reverse transcriptase-PCR technology. Results: Of the 56 malig-nant pleural effusion samples, 39 showed loss of WWOX exons 6-8 mRNA transcript (69.6%). This deletion was not detected in the 20 benign pieural effusion samples. Conclusion: WWOX gene may play an important role in the develepment of malignant Neural effusion, Detection of WWOX exons 6-8 mRNA may serve as a candidate molecular target for treatment of malignant pleural effusion and can be a promising index in differen-tial diagnosis of benign and malignant pleural effusion.

Aim To investigate whether the mechanism of curcumin antagonizing tumor correlates with histone acetylation/deacetylation,which is regarded as modulation of transcription level.Methods Measure the survival rates of Raji by treatment with various concentration of curcumin or TSA by the method of MTT;Measure the expression of acetylated histone_3 in Raji,HL60 and K562 when treated with various concenration of curcumin or TSA by means of Immunohistochemistry and FACS.Results Curcumin could inhibit Raji cell proliferation in a time-dependent and dose-dependent manner.Curcumin at the concertration of 25 ?mol?L~(-1) could enhance acetylated histones in the three tumor cells(P

Objective To characterize the systolic torsion in dilated cardiomyopathy (DCM) by velocity vector imaging(VVI).Methods Eighty-seven subjects were studied using VVI:27 patients with DCM and 60 healthy control subjects.Left ventricular short-axis acoustic images were acquired at base and apex levels.The rotation angle and rotation velocity of endocardium and epicardium were measured.Results LVEF of DCM group was significantly lower than that of control group ( P＜0.01).The basal and apical rotation angle, rotation velocity were significantly lower in DCM group.The endocardial and epicardial rotation angle, rotation velocity were also significantly lower in DCM group than those in control group (P＜0.01).Conclusions VVI is a rapid and noninvasive tool to quantitatively assess cardiac torsional deformation in DCM patients,which providing another useful modality for evaluating cardiac function.

ObjectiveTo explore the efficacy of sevoflurane or propofol combined with remifentanil during the maintenance of general anesthesia in thyroid gland surgery.MethodsSixty patients underwent thyroid gland surgery were randomly divided into tow groups.Once the larynx mask was intubated, anesthesia was maintained with propofol(the effect site concentration was 2.5 ～3.5mg/L) and remifentanil(the effect site concentration was 4.5 ～5.5μg/L) by TCI technique in group P,but with sevoflurane(2％～4％) and remfentanil(the effect site concentration was 2.5 ～4.0g/L)in group S.The depth of anesthesia was measured by the A-line TM AEP Monitor which expressed as A-Line ARX Index TM(AAI).All patients' SBP,DBP and HR were recorded at eight time points: before induction time(T0) ,after induction but before larynx mask intubation time(T1) ,intubate larynx mask time(T2) ,cut skin time (T3), separate thyroid gland time (T4), cut thyroid gland time (T5), remove larynx mask time (T6) ,leave the operation room time(T7).The emergency time,the conscious of the patients after anesthesia and the side effects were also recorded.ResultsThere were no significant differences between the groups with respect to age,gender,weight,the duration of operation,the emergency time and the conscious of the patients after anesthesia.SBP,DBP,HR of the patients in both groups showed no significant difference at T0,T1 ,T2, T3 (all P ＞ 0.05), but had significant difference at T4,T5,T6, T7 (all P ＜ 0.05).Compare with group P,the incidentce of restlessness, dizziness, drowsiness, rigor and pain was significantly lower in group S(all P ＜0.05).The incidentce of nausea,vomit and aspiration did not appear in both groups.ConclusionBoth groups showed good anesthesia effects and the patients also emerged from anesthesia quickly.But the anesthesia maintained with sevoflurane and remifentanil could bring more stable hemodynamics and lower incidence rate of the side effects.

Objective To compare the effects of patient's tolerance to laryngeal mask airway (LMA) and tracheal tube (TT) on the appropriate level of sevoflurane anesthesia.Methods Eighty ASA Ⅰ or Ⅱ patients aged 30-60 yr weighing 50-70 kg undergoing elective thyroid or breast surgery were randomly divided into 2 groups (n=40 each):LMA group and TT group.Anesthesia wag induced with propofol 1.6mg/kg,fentanyl 3μg/kg and recuronium 0.6mg/kg.LMA or tracheal tube was inserted,and the patients were mechanically ventilated.Anesthesia was maintained with inhalation of 1.5%-3.0% sevoflurane and 50% N2O in O2 in group LMA,and with 2.5%-5.0% sevoflurane and 50% N2O in O2 in group TT.The flow of O2 and N2O was 0.7-1.0L/min,and the concentration of sevofluranee was adjusted to maintain AAI at 15-25.ECG,HR,MAP,SpO2,PETCO2,AEP and end-tidal sevoflurane concentration were continuonsly monitored.The end-tidal sevoflurane concentration was recorded at 1,5 and 10 min after LMA/TT was placed (T1-3),at 1h after skin incision (T4) and at the end of operation before removal of LMA or extubation (T5).The recovery time of consciousness,adverse cardiovascular events and adverse reactions were recorded.Results The end-tidal sevoflurane concentration was significantly lower,the recovery time of consciousness and removal of LMA or extubation time were shorter,and the incidence of adverse cardiovascular events and adverse reactions was lower in LMA group than in TT group.Conclusion At the same depth of anesthesia (AAI 15-25),sevoflurane concentration is significantly lower in LMA group than in TT group,with fewer complications and smaller cardiovascular reaction.

AIM:To investigate the inhibitory effects of triptolide on cell proliferation and metastasis in Burkitt's lymphoma cell line Raji cells.METHODS: The effects of triptolide on the growth of Raji cells were studied by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium(MTT) assay.The effects of triptolide on the cell apoptosis of Raji cells were detected by using Annexin Ⅴ/PI double-labled cytometry.The effects of triptolide on CXCR4 expression on Raji cells were studied by flow cytometric analysis.Chemotaxis assays were performed to observe the effects of triptolide on migration of Raji cells towards recombinant human SDF-1?(rhSDF-1?)in vitro.RESULTS: Triptolide inhibited the proliferation of Raji cells in a dose-and time-dependent way with a 24 h IC50 value of 43.06 nmol/L and a 36 h IC50 value of 25.08 nmol/L.Following the treatment of triptolide,the cell apoptosis rate was increased as the treatment concentration increased and the culture time extended.The effects were dose-and time-dependent.Triptolide could downregulate the expression of CXCR4 on Raji cells in a dose-dependent manner.Moreover,chemotaxis assay suggested that triptolide could block the migration of Raji cells to rhSDF-1? in vitro,and the inhibition was dose-dependent.CONCLUSION: Triptolide could inhibit the cell proliferation and induce the cell apoptosis of Raji cells.Furthermore,it could block the cell metastasis of Raji cells in vitro and the underlying mechanism might be related to the inhibition of the SDF-1/CXCR4 axis.

Overexpression of human ether-脿-go-go (eag) related gene (hERG) has been found in a broad range of human leukemia cell lines and primary human leukemia. The block of hERG protein might be a potential therapeutic strategy for leukemia. Gambogic acid (GA) has recently exhibited marked anti-tumor potency on solid tumors of various derivations. Here, we investigated the anti-leukemia effects of GA and its relation to the regulation of hERG in K562 leukemia cells in vitro. K562 cells were treated with various concentrations of GA (0.125-8.0 micromol/L) for 0-72 h. MTT assay was used to evaluate the inhibition effect of GA on the growth of K562 cells. Cell apoptosis was measured through both Annexin-V FITC/PI double-labeled cytometry and transmission electron microscopy. Cell cycle regulation was studied by a propidium iodide method. RT-PCR and Western blot were applied to detect the expression level of hERG in K562 cells. GA presented striking growth inhibition and apoptosis induction potency on K562 cells in vitro in a time- and dose-dependent manner. The IC(50) value of GA for 24 h was 2.637+/-0.208 micromol/L. Moreover, GA induced K562 cells arrested in G(0)/G(1) phase, accordingly, cells in S phase decreased gradually, and no obvious changes were found in G(2)/M phase cells. Under the transmission electron microscopy, apoptotic bodies containing nuclear fragments were found in GA-treated K562 cells. After treatment with GA of 2.0 micromol/L for 24 h, the percentage of apoptotic cells was increased from 4.09% to 18.47% (P<0.01). Overexpression of hERG channel was found in K562 cells, while GA could down-regulate it at both protein and mRNA levels (P<0.01). It was concluded that GA exhibited its anti-leukemia effects partially through down-regulating the expression level of hERG channel in K562 cells, suggesting that GA may be a potential agent against leukemia with a mechanism of blocking hERG channel.

The inhibitory effect of wortmannin on leukemic cells and the possible mechanisms were examined. K562 cells were treated with wortmannin of various concentrations (3.125-100 nmol/L) for 0-72 h. MTT assay was used to evaluate the inhibitory effect of wortmannin on the growth of K562 cells. Cell apoptosis was detected by both Annexin-V FITC/PI double-labeled cytometry and transmission electron microscopy (TEM). The expression of p-Akt, T-p-Akt, NF-kappaBp65 and IKK-kappaB was determined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that wortmannin obviously inhibited growth and induced apoptosis of K562 cells in vitro in a time- and dose-dependent manner. The IC(50) value of wortmannin for 24 h was 25+/-0.14 nmol/L. Moreover, wortmannin induced K562 cells apoptosis in a dose-dependent manner. TEM revealed typical morphological changes of apoptosis in wortmannin-treated K562 cells, such as chromatin condensation, karyopyknosis, karyorhexis and apoptotic bodies. Additionally, several important intracellular protein kinases such as p-Akt, NF-kappaBp65 and IKK-kappaB experienced degradation of various degrees in a dose-dependent manner both at protein level and transcription level when cultured with wortmannin, but the expression of total Akt showed no change. It is concluded that wortmannin can inhibit the proliferation and induce apoptosis of K562 leukemia cells possibly by down-regulating the survival signaling pathways (PI3K/Akt and NF-kappaB channels).

In order to investigate the anti-leukemia effects of gambogic acid (GA) and its relation to the regulation of nucleoporin Nup88 in U937 cells in vitro, the inhibitory effect of GA on the growth of U937 cells was examined by using MTT assay. Apoptosis was detected by Annexin-V FITC/PI double-labeled cytometry. Cell cycle regulation was studied by propidium iodide method. Both flow cytometry (FCM) and RT-PCR were employed to assess the expression of Nup88, and the localization of Nup88 was determined by confocal microscopy. The results indicated that GA had strong inhibitory effect on cell proliferation and apoptosis induction activity in U937 cells in vitro in a time-and dose-dependent manner. The 24-h IC(50) value was (1.019+/-0.134) mg/L. Moreover, GA induced arrest of U937 cells in G(0)/G(1) phase. Over-expression of Nup88 was found in U937 cells, whereas GA could significantly down-regulate both the protein and mRNA levels of Nup88. Nup88 was diffusely distributed between nucleus and cytoplasm and was located at the cytoplasmic side of nuclear rim, and occasionally in cytoplasm. It is suggested that GA exerts its anti-leukemia effects by regulating the expression and distribution of nucleoporin Nup88. It promises to be new agent for the treatment of acute leukemia.
Antineoplastic Agents, Phytogenic/*pharmacology ; Apoptosis/drug effects ; Cell Proliferation/drug effects ; Nuclear Pore Complex Proteins/genetics ; Nuclear Pore Complex Proteins/*metabolism ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; U937 Cells ; Xanthones/*pharmacology

This study explored the molecular mechanisms underlying the time-dependent autophagy and apoptosis induced by nutrient depletion in human multiple myeloma cell line RPMI8226 cells. RT-PCR and qRT-PCR were used to evaluate the transcriptional levels of Deptor, JNK1, JNK2, JNK3, Raf-1, p53, p21 and NFκB1 at 0, 6, 12, 18, 24 and 48 h after nutrient depletion in RPMI8226 cells. We found that transcriptional levels of Deptor were increased time-dependently at 0, 6, 12 and 18 h, and then decreased. Its alternation was consistent with autophagy. Transcriptional levels of Raf-1, JNK1, JNK2, p53 and p21 were increased time-dependently at 0, 6, 12, 18, 24 and 48 h accompanying with the increase of apoptosis. Transcriptional levels of NFκB1 at 6, 12, 18, 24 and 48 h were decreased as compared with 0 h. It was suggested that all the studied signaling molecules were involved in cellular response to nutrient depletion in RPMI8226 cells. Deptor contributed to autophagy in this process. Raf-1/JNK /p53/p21 pathway may be involved in apoptosis, and NFκB1 may play a possible role in inhibiting apoptosis. It remained to be studied whether Deptor was involved in both autophagy and apoptosis.