The aim of this study was to investigate the effect of bcl-2 siRNA on bcl-2 gene expression and apoptosis of lymphoma cell line CA46. A siRNA was designed and synthesized. Then siRNA was transfected into CA46 cells by cationic liposome. At 48 hours after transfection, apoptosis and mitochondria transmembrane potential of CA46 cells were detected by flow cytometry, the expression of bcl-2 mRNA and BCL-2 protein in CA46 cells were detected by RT-PCR and flow cytometry respectively. The results showed that at 48 hours after transfection, apoptosis of CA46 cells occurred, mitochondria transmembrane potential changed. The expression of bcl-2 mRNA and BCL-2 protein in CA46 cells decreased significantly. In conclusion, bcl-2 siRNA depresses the expression of bcl-2 gene in CA46 cells specifically, then changes the mitochondria transmembrane potential, resulting in apoptosis of CA46 cells.
Apoptosis ; genetics ; Burkitt Lymphoma ; genetics ; Cell Line, Tumor ; Genes, bcl-2 ; Humans ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

This study is to investigate the effect of emodin on inducing human myeloid leukemia cell line HL-60 apoptosis and the role of Akt signal pathway in the apoptosis. HL-60 cells were exposed to various dosages of emodin. MTT assay was used to detect HL-60 cell proliferation. Distribution of HL-60 cells in cell cycle was analyzed by flow cytometry and cell apoptosis was observed by MitoCapture apoptosis detection. The protein expressions of Akt signal pathway were detected by Western blotting. The result showed that emodin remarkably inhibited the cell proliferation. The IC50 value for 48 h treatment was about 20 micromol x L(-1). Apoptosis in HL-60 cells could be efficiently induced by emodin in a dose dependent manner and cells were arrested at G0/G1. The expressions of Akt, p-Akt, IkappaB-alpha, p-IkappaB-alpha, p65, p-p65, mTOR and p-mTOR in Akt signal pathway were downregulated after emodin treatment. It can be concluded that emodin could efficiently induce growth inhibition and apoptosis in HL-60 cells. Akt signal pathway may be involved in this process.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Emodin ; pharmacology ; HL-60 Cells ; Humans ; I-kappa B Proteins ; metabolism ; NF-KappaB Inhibitor alpha ; Protein Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; Transcription Factor RelA ; metabolism

OBJECTIVETo study the effects of bcl-2 antisense phosphorothioate oligonucleotides (ASPO) on suppression of HL-60 cell growth in SCID mice and to investigate the feasibility of purging leukemia cells plus bcl-2 ASPO used in vitro.

METHODS1 x 10(7) viable HL-60 cells were treated with 10 micro mol/L bcl-2 ASPO seven days before the intraperitoneal (IP) inoculation to the SCID mice, Treatment with sense oligonucleotides (SPO) was similar as for the controls. 35 days after the inoculation, all the SCID mice of both groups were sacrificed and their peripheral blood, bone marrow, liver and spleen were examined using half nested RT-PCR and histopathology for detecting the appearance and distribution of the HL-60 cells treated beforehand with antisense or sense oligonucleotides respectively.

RESULTSASPO could down regulate the expression of bcl-2 resulting in both inhibition of growth and induction of apoptosis in treated HL-60 cells, which failed to develop leukemia in SCID mice at all. However, SPO treated HL-60 cells still behaved their own ways and proliferated agressively, and developed leukemia at last.

CONCLUSIONThe bcl-2 ASPO enables to suppress HL-60 cell growth and prevent the development of leukemia in the SCID mice. The purging leukemia cells used are seemed liable in inhibiting the development of leukemia in SCID mouse model.

Animals ; Cell Division ; drug effects ; Disease Models, Animal ; HL-60 Cells ; drug effects ; Humans ; Mice ; Mice, SCID ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; genetics

OBJECTIVETo study the clinicopathologic features, diagnostic criteria, differential diagnosis and treatment options of ovarian steroid cell tumor, not otherwise specified (NOS).

METHODSLight microscopy and immunohistochemical study was carried out in 8 cases of ovarian steroid cell tumor, NOS. The literature was reviewed.

RESULTSThe 7 cases of benign ovarian steroid cell tumor, NOS were composed mainly of polygonal cells with granular eosinophilic cytoplasm and larger cells with vacuolated cytoplasm. They resembled the architecture of normal adrenal gland, with formation of cell nests and trabeculae. The single case of malignant ovarian steroid cell tumor had evidence of significant cellular pleomorphism, haemorrhage and coagulative tumor necrosis. The mitotic count measured about 7 per 10 high-power fields. Immunohistochemical study showed that the tumor cells expressed calretinin and alpha-inhibin. Differential diagnosis included oxyphilic granulosa cell tumor, thecoma, Sertoli cell tumor and clear cell carcinoma. The treatment options of benign ovarian steroid cell tumor, NOS was local excision or ipsilateral salpingo-oophorectomy, while the malignant counterpart should be treated with a combination of surgery and chemotherapy, including administration of GnRH agonist.

CONCLUSIONSOvarian steroid cell tumor, NOS, is the most common type of ovarian steroid cell tumors. Most of which are associated with a benign clinical outcome. Immunohistochemistry is an important adjunct for diagnosis. The treatment options of ovarian steroid cell tumor, NOS depend on its malignant potential.

Adolescent ; Adult ; Calbindin 2 ; Diagnosis, Differential ; Female ; Granulosa Cell Tumor ; pathology ; Humans ; Inhibins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Ovariectomy ; methods ; Ovary ; pathology ; S100 Calcium Binding Protein G ; metabolism ; Sertoli Cell Tumor ; pathology ; Sex Cord-Gonadal Stromal Tumors ; metabolism ; pathology ; surgery ; Thecoma ; pathology ; Young Adult

The fused tooth is the union of two dental enamel or dentin formed together. In the maxillary, the fusion usually occurred within the lateral incisor and canine and very rarely occurred in the upper third molar and supernumerary tooth. This paper reported a fused tooth occurred in the left maxillary impacted third molar with supernumerary tooth.
Fused Teeth ; Humans ; Incisor ; Male ; Maxilla ; Molar ; Molar, Third ; Tooth, Impacted ; Tooth, Supernumerary

To investigate the change of telomerase activity and human telomerase reverse transcriptase (hTERT) gene expression in HL-60 cells transfected with wild type p53 gene, wild type p53 gene was introduced into HL-60 cells by Lipofectin transfection. Apoptosis was analyzed by TUNEL assay. Telomerase activity and the level of hTERT mRNA were detected by telomeric repeat amplification protocol (TRAP)-ELISA and RT-PCR, respectively. The results showed that the apoptotic rate of HL-60-pN53cG cells was 8.3% and 21.0% respectively after cultured at 32.5 degrees C for 24 h and 72 h. The level of hTERT mRNA was decreased to 68.4% and 55.8% and telomerase activity to 27.3% and 8.9% of control value in HL-60-pN53cG cells at the same points. In conclusions, hTERT mRNA and telomerase activity were down-regulated in HL-60 cells transfected with p53 gene. This may be one of mechanisms of apoptosis induced by wild type p53 gene.
Apoptosis ; DNA-Binding Proteins ; Gene Expression ; Genes, p53 ; physiology ; HL-60 Cells ; Humans ; RNA, Messenger ; analysis ; Telomerase ; genetics ; metabolism

The study was aimed to investigate the effects of emodin on the proliferation and apoptosis of adriamycin-resistant HL-60/ADR cells, and to explore the underlying mechanism. The cell viability and colony formation were detected by MTT assay and colony formation assay respectively. Apoptotic cells were tested by means of cell cycle analysis, mitochondrial transmembrane potential levels, caspase-3 activity detection, Annexin V FITC/PI staining and TUNEL labeling. RT-PCR was used to analyze the bcl-2 and c-myc mRNA expressions. The protein expressions of Bcl-2, c-Myc and caspase-3 precursor were determined by Western blot. The results showed that HL-60/ADR cell growth was significantly inhibited by emodin in dose and time dependent manners. Cell colony formation obviously decreased with IC50 5.79 micromol/L. G0/G1 phase cell population increased while G2/M phase cells decreased in 40 and 80 micromol/L groups compared with control group (p < 0.01), and no significant difference of cell cycle was observed in 20 micromol/L group (p > 0.05). The typical hypo-diploid peak (apoptotic peak) appeared in each dose group. The levels of mitochondrial transmembrane potential of HL-60/ADR cells decreased and caspase-3 activity increased when incubated with emodin for 12 and 24 hours respectively. Apoptosis occurred in a dose-dependent manner, and its earlier and later stages were identified by Annexin-V FITC/PI staining and TUNEL labeling methods respectively. The expressions of bcl-2, c-myc mRNA and Bcl-2, c-Myc, caspase-3 precursor protein were all down-regulated in a time-dependent manner after treatment with emodin at different times. It is concluded that emodin efficiently inhibits growth and induces apoptosis on HL-60/ADR cells, which may be related with the down-regulation of mitochondrial transmembrane potential and expressions of bcl-2 and c-myc, as well as up-regulation of caspase-3 activity.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; drug effects ; Emodin ; pharmacology ; HL-60 Cells ; Humans

OBJECTIVETo evaluate the evolution of medically treated atherosclerotic aortic ulcers by computed tomography (CT).

METHODSThirty-five patients (31 men and 4 women, aged from 40 to 79 years, mean 56.2 +/- 10.8 years) with known aortic ulcers were monitored by CT (follow up time 7 - 730 days, mean 135 days), 80 - 100 ml contrast media (Ultravist 300 or 320, or Omnipaque 300 or 320 mg/ml) was injected with a rate of 3.5 - 4.5 ml/s. The scan delayed time was 18 - 30 s. Ulcers dimensions were measured according to maximum depth, maximum length and maximum width.

RESULTSThirty-one patients with intramural hematomas and 1 patient with atherosclerotic aortic arch aneurysm without intramural hematoma were medically treated and another 3 patients were surgically treated. Intramural hematoma regression was monitored in 31 medically treated patients with intramural hematomas. CT was repeated at 2 weeks, 3 and 6 months. Intramural hematoma resolved gradually during follow up [thickness: (7.69 +/- 4.24) mm at 3 months, (3.06 +/- 1.67) mm at 6 months, P < 0.05 vs. 1st CT: (11.96 +/- 4.16) mm while ulcer maximum depth (11.17 +/- 6.03) mm at 3 months, (11.35 +/- 5.59) mm at 6 months, P < 0.05 vs. 1st CT: (7.36 +/- 6.61) mm, maximum width (14.40 +/- 6.35) mm at 3 months, (18.55 +/- 10.94) mm at 6 months, P < 0.05 vs. 1st CT: (7.15 +/- 6.39) mm, maximum length (17.12 +/- 7.15) mm at 3 months, (18.13 +/- 10.89) mm at 6 months, P < 0.05 vs. 1st CT: (11.64 +/- 10.06) mm increased progressively during follow-up].

CONCLUSIONCT was a useful tool for deflecting atherosclerotic aortic ulcers and monitoring therapeutic effects.

Adult ; Aged ; Aortic Diseases ; diagnostic imaging ; Aortography ; Atherosclerosis ; diagnostic imaging ; Female ; Follow-Up Studies ; Hematoma ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Ulcer ; diagnostic imaging

The aim of study was to investigate the effect of a traditional Chinese medicine, emodin, on proliferation and apoptosis in T lymphocytic leukemic cell line Jurkat and its mechanisms. Cell proliferation inhibition was detected by MTT assay. Cell apoptosis was measured by DNA ladder and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The expressions of related proteins and caspase family members were determined by Western blot. The results showed that emodin inhibited proliferation in Jurkat cells, with an IC50 about 20 micromol/L and induced cell apoptosis in both time-and dose-dependent manners. The expressions of proliferation-related protein C-MYC, hTERT and apoptosis-related protein BCL-2 were down-regulated in a time dependent manner after the treatment with emodin. The expressions of procaspase-3, -8 and -9 all decreased while activated caspase-3 and PARP expressions were up-regulated. It is concluded that emodin can remarkably inhibit cell proliferation and induce apoptosis in Jurkat cells. The down-regulation of proliferation-related proteins C-MYC, hTERT and apoptosis-related protein BCL-2 expressions and activation of caspase cascade may be involved in the process of apoptosis.
Apoptosis ; drug effects ; Caspases ; metabolism ; Cell Proliferation ; drug effects ; Emodin ; metabolism ; pharmacology ; Humans ; Jurkat Cells

OBJECTIVETo study the morphological features of secundum atrial septal defect (ASD) in adult and the implications for transcatheter closure.

METHODSTranscatheter closure using Amplatzer duct occluder was performed in 272 adult patients with ASD from September 1997 to December 2005. The morphological features were evaluated by transthoracic echocardiography (TTE) and transesophageal echocardiography (TEE). The size, length and thickness of rims, occluder diameter, the complete closure rate, residual shunt rate and complications were compared in patients with deficient and/or thin rims (Group A, n = 135) and patients with well-developed rims (Group B, n = 137).

RESULTSThe complete closure rate was 97.8% (132/135) in group A and 99.3% (136/137) in group B. There were 74 cases with deficient rims, 39 cases with thin rims and 22 cases with both deficient and thin rims in group A. Gender distribution, age, operation successful rate, residual shunt rate and complication rate were similar between the 2 groups. The defect diameters measured by TTE (18.9 +/- 5.5 mm vs. 16.5 +/- 4.8 mm, P < 0.01), TEE (22.7 +/- 5.0 mm vs. 20.0 +/- 5.5 mm, P < 0.01) and occluder diameters used (29.1 +/- 5.7 mm vs. 26.0 +/- 5.9 mm, P < 0.01) were significantly larger in groups A than that in group B. The systolic pulmonary artery pressure was also significantly higher in groups A than that in groups B (36.9 +/- 11.9 mm Hg vs. 32.6 +/- 9.1 mm Hg, P < 0.01). There are significant correlations between occluder diameters and defects measured by either TTE or TEE in both groups (group A, TTE: r = 0.709, TEE: r = 0.850; group B, TTE: r = 0.716, TEE: r = 0.915, P all < 0.01).

CONCLUSIONSPoor residual rims were found in around 50% of adult patients with ASD. Transcatheter closure of these defects could be successfully performed with larger occluders. The defect diameters measured by TTE and TEE, especially the latter, could guide the occluder selection.

Adult ; Cardiac Catheterization ; Female ; Follow-Up Studies ; Heart Septal Defects, Atrial ; etiology ; pathology ; therapy ; Humans ; Male ; Middle Aged