OBJECTIVETo investigate the effect of phosphorylated protein kinase C epsilon (pPKC epsilon) on apoptosis of 32D cells induced by sera from patients with aplastic anemia (AA).

METHODSThe expression of pPKC epsilon and apoptosis in 32D cells were measured by Western blotting and flow cytometry after incubation with sera from healthy individuals (controls, n = 8), patients with severe AA ( SAA, n = 8)and non severe AA (NSAA, n = 6).

RESULTSAfter incubation for 0, 12, 24, 36 and 48 hours in the presence of serum and for another 4 hours in medium deprived of serum, the levels of pPKC epsilon in cells in SAA and NSAA group increased gradually, peaked at 24 hours, and then declined (P < 0.05). Compared with that in control group (0.54 +/- 0.08), pPKC epsilon was overexpressed in both SAA group (0.90 +/- 0.10) and NSAA group (0.64 +/- 0.08) (P < 0.05) after 24 hours incubation with serum and subsequent 4 hours without serum. pPKC epsilon level was higher in SAA group than in NSAA group (P < 0.05). A greater proportion of 32D cells showed apoptosis after 24 hours incubation with sera from SAA patients [(4.05 +/- 1.05)%] and subsequent 4 hours incubation without serum than that in controls [(2.45 +/- 0.51)%, P < 0.05], which was correlated with the same serum-induced expression of pPKC epsilon (r = 0.869, P < 0.05). Although the mean level of pPKC epsilon expression was higher in NSAA group than in control group, no significant difference of apoptosis was found between the two groups [(2.45 +/- 0.51)% vs (3.24 +/- 0.56)%, P > 0.05].

CONCLUSIONSera from both SAA and NSAA patients could upregulate the expression of pPKC epsilon in 32D cells. The SAA sera induce apoptosis in 32D cells significantly, but the latter do not.

Adolescent ; Adult ; Anemia, Aplastic ; enzymology ; pathology ; Apoptosis ; Case-Control Studies ; Cells, Cultured ; Child ; Female ; Humans ; Male ; Middle Aged ; Phosphorylation ; Protein Kinase C-epsilon ; blood ; Young Adult

This study was aimed to investigate the expression of vascular endothelial growth factor (VEGF) in patients with aplastic anemia. The gene expressions of VEGF in mononuclear cells of bone marrow from 7 cases of aplastic anemia and 12 normal controls were detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The expressions of VEGF in bone marrow from 20 cases of aplastic anemia and 20 normal controls were also determined by immunohistochemistry assay. The results showed that expression of VEGF mRNA was found in 2 out of 7 (28.57%) bone marrow of patients and in 10 out of 12 (83.33%) bone marrow of normal controls. The VEGF mRNA in patients with aplastic anemia was significantly lower than that in normal controls (P < 0.05). No patients with aplastic anemia showed immunohistochemical staining of VEGF in bone marrow, while 5 out of 20 (25%) normal controls exhibited VEGF positive cells. Bone marrow of aplastic anemia patients contained less VEGF than that of normal persons (P < 0.05). In conclusion, when compared with normal controls, VEGF expression decreased significantly in patients with aplastic anemia at gene transcription level and protein translation level, it may be related to the defect of angiogenesis and thus hematopoiesis in bone marrow of patients with aplastic anemia.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; metabolism ; Base Sequence ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

The aim of this study was to investigate the inhibition effect of arsenic sulfide (As2S2) on the growth of in vitro cultured BMMNC from MDS patients and to explore its possible cellular and molecular mechanisms. The apoptosis of MDS cells induced by As2S2 solution of different concentrations were studied with MTT, flow cytometry, and RT-PCR. The results showed that (1) low concentration of As2S2 (0-0.6 mg/L) had no marked inhibition effect on proliferation of MDS cells; (2) after treatment with 1.5-50 mg/L of As2S2, both low risk MDS cells and high risk MDS cells presented typical features of apoptosis with a dose-dependent manner, the expression of bcl-2 mRNA and the ratio of bcl-2/bax obviously decreased after As2S2 treatment (P < 0.05); (3) BMMNC from MDS patients had higher apoptosis ratio than that of BMMNC from control. It is concluded that BMMNC excessive apoptosis exists in MDS patients; low concentration of As2S2 (0-0.6 mg/L) shows no inhibition effect on proliferation of MDS cells; high concentration of As2S2 (1.5-50 mg/L) induces apoptosis of MDS cells.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Bone Marrow Cells ; pathology ; Cyclin D1 ; biosynthesis ; genetics ; Female ; Humans ; Leukocytes, Mononuclear ; pathology ; Male ; Middle Aged ; Myelodysplastic Syndromes ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Sulfides ; pharmacology ; bcl-2-Associated X Protein ; biosynthesis ; genetics

OBJECTIVETo investigate the down-regulated TRAF6 gene expression and its effects on proliferation and apoptosis in multiple myeloma (MM) cells.

METHODSDetection of TRAF6 expression were conducted by RT-PCR and Western blot in MM cell lines of KM3, U266, RPMI8226 and primary cells from patients. RPMI8226 cell lines were transfected with siRNA of TRAF6. The efficiency of transfection was identified by using of fluorescence microscope, RT-PCR, and Western blot. The levels of proliferation were analyzed by CCK-8 method under the different concentrations of siRNA. Apoptosis rate were detected with Hoechst33258/PI double staining by flow cytometry. Apoptosis related proteins Bcl-2, BAX, and NF-κB signal pathway were observed before and after siRNA transfection by Western blot.

RESULTSThe levels of TRAF6 mRNA and protein in MM cell lines, especially in primary myeloma cells, were significantly higher than those in controls. After transfected with 50 nmol/L siRNA in RPMI8226 cells, the relative level of TRAF6 mRNA (0.49±0.24) was significantly lower than that in non-transfected group (1.87±0.23) and idling group (1.74±0.35). The proliferation rate of siRNA transfected cells decreased with dose dependence (P<0.01). The apoptosis rates increased from 11.20% (before transfection) to 51.82% (after transfection), accompanied by down-regulated Bcl-2 protein, NF-κB signal pathway (p-p65 and p52), and up-regulated BAX protein.

CONCLUSIONTRAF6 expression was high in myeloma cells. TRAF6 siRNA could inhibit proliferation of myeloma cells and induce apoptosis mediated by NF-κB classical and alternative pathway in myeloma cells.

Case-Control Studies ; Cell Proliferation ; Down-Regulation ; Female ; Gene Expression ; Humans ; Male ; Multiple Myeloma ; metabolism ; pathology ; TNF Receptor-Associated Factor 6 ; genetics ; metabolism ; Tumor Cells, Cultured

To explore the difference of negative regulatory factors among T lymphocyte subsets in bone marrow (BM) of myelodysplastic syndromes (MDS) and their relations to apoptotic gene Fas, different lymphocyte subsets in BM were categorized by monoclonal antibodies with 3 color fluorescence using flow cytometry, and the intracellular cytokines such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) were determined following marrow cells culture. Then Fas mRNA of bone marrow mononuclear cells (BMMNC) were examined by RT-PCR. The results showed that TNF-alpha, IFN-gamma levels in BM of MDS both increased, the former produced by cells CD4+CD45RO+, CD8+CD45RO+, the latter by cells CD4+CD45RO+, CD8+CD45RO+, CD8+CD45RA+, in which the cells CD8+CD45RO+ were dominant. Fas mRNA expression had relationship with IFN-gamma produced by T cells but not with TNF-alpha. It is concluded that in hematopoietic microenvironment of MDS, not only the T lymphocyte subsets are in disorder, but also negative regulatory factors secreted by T lymphocyte increase. T lymphocytes play an important role in producing IFN-gamma in patients with MDS.
Adult ; Aged ; Bone Marrow Cells ; metabolism ; Female ; Humans ; Interferon-gamma ; biosynthesis ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; RNA, Messenger ; analysis ; T-Lymphocyte Subsets ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; fas Receptor ; genetics

OBJECTIVETo study the inhibitory effect of zoledronic acid (ZA) on the growth and CD138 expression of myeloma cell line KM3.

METHODSKM3 cells were treated with different concentrations of ZA The growth of KM3 cells was measured by trypan blue dye exclusion, and the changes of apoptosis rate, cell cycle and expression of CD138 induced by ZA by flow cytometry.

RESULTSWithin the concentration of 10(-5)-10(-3) mol/L, ZA obviously inhibited the growth of KM3 cells in a dose dependent manner. IBN at 10(-5)-10(-4) moL/L increased Annexin V positive rate, blocked cells at the S/G2 boundary, reduced the expression of CD138 and its fluorescence intensity.

CONCLUSIONZA can inhibit the growth of KM3 cells in a dose-dependent manner and inhibited CD138 expression. The mechanism is probably related to induction cell cycle accumulation in S phase and apoptosis.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diphosphonates ; pharmacology ; Humans ; Imidazoles ; pharmacology ; Syndecan-1 ; metabolism

Objective To investigate the Wallerian degeneration of neural fiber tract in medulla and bilateral middle cerebellar peduncle following pontine infarction and to explore its impacts on neurological recovery.Methods Fourteen patients with a recent unilateral pontine infarct underwent the diffusion tensor imaging(DTI)and evaluations with the NIH stroke scale(NIHSS),the Fugl-Meyer motor scale(FM), ataxia rating scale(ARS)and the Barthel index(BI)at the first week(W1),the fourth(W4)and twelfth week(W12)respectively.Mean diffusivity(MD)and fractional anisotropy(FA)were measured at pons, medulla and middle cerebellar peduncle.Fourteenth age and gender matched volunteers underwent a DTI were studied as controls.Results Compared with the matched regions in controls,the FA values of infarct side medulla and bilateral middle cerebellar peduncle in patients significantly decreased at W1,W4 and W12(on the infarct side of medulla:W1:0.43?0.01;W4:0.37?0.02;W12:0.30?0.02;on the infarct side of middle cerebellar peduncle:W1:0.50?0.01;W4:0.43?0.02;W12:0.35?0.04;on the opposite side of middle cerebellar peduncle infarction:W1:0.54?0.02;W4:0.52?0.03;W12:0.47?0.04,t values are 1.92 to 28.56,P0.05 respectively).The absolute value of percent reduction of FA in infarct side medulla and bilateral middle cerebellar peduncle was correlated negatively to the absolute value of percent change of NIHSS and BI score(P

OBJECTIVETo assess the safety and feasibility of laparoscopic and open repair of perforated peptic ulcer.

METHODSStudies on comparison between laparoscopic repair(LR) and open repair(OR) of perforated peptic ulcer were collected. Data of operating time, blood loss, time to first flatus, postoperative hospital stay, postoperative complications and mortality between LR group and OR group were meta-analyzed using fixed effect model and random effect model.

RESULTSNineteen studies including 1507 patients were selected for this study,including laparoscopic surgery(n=673) and open surgery(n=834). There were significant differences in blood loss, time to first flatus, postoperative hospital stay, wound infection rate and mortality between LR group and OR group. However, no significant differences existed in operative time, postoperative sepsis, pulmonary infection, abdominal abscess, and suture leakage between the two groups.

CONCLUSIONSLaparoscopic repair of perforated peptic ulcer is associated with improved outcomes in terms of less blood loss, quicker recovery, and lower rates of wound infection and mortality. Laparoscopic repair of perforated peptic ulcer is safe and feasible.

Humans ; Laparoscopy ; Laparotomy ; Peptic Ulcer Perforation ; surgery ; Treatment Outcome

BACKGROUNDTransradial access has been increasingly used during primary percutaneous coronary intervention (PCI) for patients with acute ST-segment elevation myocardial infarction (STEMI) in last decade. Clinical benefits of upstream use of tirfiban therapy in STEMI patients treated by primary PCI have been reported. We investigated the merits of transradial vs. transfemoral access in primary PCI for STEMI patients with upstream use of tirofiban.

METHODSPatients with STEMI treated with tirofiban between December 2006 and October 2012 then by primary PCI were compared between transradial (n = 298) and transfemoral (n = 314) access. Baseline demographics, angiographic and PCI features and primary endpoint of major adverse cardiac events (MACE) at 30-day clinical follow-up were recorded.

RESULTSBaseline and procedural characteristics were comparable between the two groups, apart from more patients in transradial group had hypertension and were treated by thrombus aspiration during primary PCI. Significantly fewer MACE occurred in the transradial group (5.4%) compared with the transfemoral group (9.9%) at 30-day clinical follow-up. Major bleeding events at 30-day clinical follow-up were 0 in transradial group and in 2.9% of transfemoral group. Multivariate analysis confirmed transradial approach as an independent negative predictor of 30-day MACE (HR 0.68; 95%CI 0.35 - 0.91; P = 0.03).

CONCLUSIONSUsing transradial approach in primary PCI for acute STEMI infarction patients treated with tirofiban was clearly beneficial in reducing bleeding complications and improving 30-day clinical outcomes.

Aged ; Angioplasty, Balloon, Coronary ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; drug therapy ; therapy ; Percutaneous Coronary Intervention ; methods ; Tyrosine ; analogs & derivatives ; therapeutic use

Objective The impact of late incomplete stent apposition(ISA)post sirolimus eluting stent(SES)implantation in patients with acute coronary syndrome(ACS)on long-term clinical outcomes remainB controversial.The alm of the present study was to eva]uate the association between late ISA and clinical outcomes in patients with ACS compared with that with stable angina(SA).Methods From February 2005 to March 2007,54 ACS patients and 83 SA patients were enrolled in this study,late ISA was determined by meallS of threc-dimensional volumetric intravaaculair ultrasound(IVUS)analyses one year after SES implantation and clinical outcomes one year post IVUS were obtained in these patients.Results In 219 treated lesions of the 137 patients,late ISA wag documented in 25 lesions in 16 patients(20 ISA in 12 ACS patients vs.5 ISA in 4 SA patients,P＜0.001).Thoush lunlen area in reference and stented segment,neointimal hyperplasia(NIH)area and percentage of NIH in stented segment,and external elastic membrane(EEM)area in reference segment were similar between two groups,EEM area in stented segment [(15.34±5.44)mm2vs.(13.83±4.51)mm2,P=0.026],stented/reference segment EEM area ratio (1.13±0.22 vs.1.02±0.18,P＜0.001),plaque and media area[(8.43±3.93)mm2vs.(7.01±2.93)mm2,P=0.002]was significantly lager in ACS group than that in SA group.Multivariable logistic analysis showed that ACS(OR 6.477 with 95%CI from 2.297 to 18.263,P＜0.001)and stent length≥23 mm(OR 3.680 with 95% CI from 1.181 to 11.469,P=0.025)were main independent factors of occurrence of late ISA.Incidence of main adverse cardiac events(MACE)one year post IVUS Wag similar between the two groups.Conclusion Compared with patients with SA,ACS patients had larger stented segment EEM area,plaque and media areaas well as increased incidence of ISA.However,the incidence of MACE was similar in ACS and SA patients one year after IVUS.