Objective To explore the effects and the mechanism of manumycin on KAT-18 cells Methods Human ATC (anaplastic thyroid cancer) KAT-18 cells were used. The cytotoxicity was analyzed by SRB assay. Apoptosis and cellular nitric oxide were detected by flow cytometry using annexin v and NO sensor dye. Superoxide anion was measured with a fluorescent plate reader by DHE.GSH was assayed by fluorescent Monochlorobimane. The SOD activities were assayed by colorimetric methods. The protein expression of Mn-SOD was determined by western blot. Results Manumycin decreased the viability of KAT-18 cells in a dose-dependent manner. Manumycin induced apoptosis significantly and NO generation simultaneously. Manumycin also induced superoxide anions generation. Manumycin reduced intracellular GSH in a time-course manner. However , manumycin did not decrease the SOD activity after 6 h treatment and Mn-SOD expression. A delayed induction of SOD activity was observed after 24 h manumycin treatment. N-acetyl-L-cysteine blocked NO and superoxide anions generation and apoptosis induction by Manumycin. Furthermore , NAC protected KAT-18 cells from the cytotoxicity of manumycin. Conclusion Manumycin induces apoptosis and has cytotoxic effects on KAT-18 cells. Cellular NO and superoxide anions generation are required for Manumycin-induced apoptosis in KAT-18 cells.

Background Fractalkine is involved in the impairment of endothelium by mediating inflammatory cell chemotaxis.Aspirin inhibites many kinds of cytokine expression.Objective To investigate the effect of aspirin on tumor necrosis factor ?(TNF-?) stimulated fractalkine expression in human umbilical vein endothelial cells (HUVEC) and its mechanism.Methods HUVEC were grouped as follows:control group;TNF-?-stimu- lated;TNF-?+NS-398;TNF-?+PDTC and TNF plus various concentration of aspirin (0.02,0.2,1.5 mmol/L). The level of mRNA and protein expression of fractalkine and nuclear factor(NF)-kB p65 was determined by RT- PCR and Western blot.Results 1)Fractalkine mRNA and protein level was increased after 4 ?g/L TNF-? stim- ulation in HUVEC(both P

Objective To investigate infection situation and knowledge of high‐risk human papilloma virus(HPV)among women working in entertainment places in Gaoqiao area ,and provide theory evidences for making health education measures related to high‐risk HPV .Methods According to the systematic random sampling method ,entertainment female in Gaoqiao area were extracted , and normal women who participated in community gynecology census were extracted as control group .Then questionnaire investiga‐tion and laboratory testing were carried out .Results The infection rate of high‐risk HPV in entertainment female was 35 .18% , while in normal woman was 15 .84% ,there was significant differences of infection rate between the two groups (P<0 .05) .There were were only 16 women(14 .81% )and 17 women(16 .83% )had heard of information about HPV in two groups respectively ,and there was no statistically significant difference(P>0 .05) .The cognitive situation of whether a person with family history of cervi‐cal cancer should be regularly attend screening among the two groups of women was different ,and had significant difference(P<0 .05) .Conclusion Women who worked in entertainment places had higher infection rate of high‐risk HPV ,both of the two groups had poor knowledge of HPV .So the propaganda and screening of HPV should be strengthen in order to raise consciousness in pre‐venting cervical cancer .

Adenoviridae ; genetics ; Calcium-Binding Proteins ; genetics ; metabolism ; Cells, Cultured ; Female ; Genetic Vectors ; Humans ; Microfilament Proteins ; genetics ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Myometrium ; cytology ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Objective To investigate the correlation between serum homocysteine (Hcy) and macrophage migration inhibitory factor(MIF) with carotid atherosclerosis and between serum Hcy and MIF ,and to study whether serum Hcy influence the carotid atherosclerosis formation by MIF .Methods 258 inpatients and outpatients were performed the color ultrasound examination to ob-serve the carotid arterial vascular anatomic form ,endomembrane circumstance ,plaque and the plaque echo nature ,and the carotid in-tima-media thickness(IMT) was measured .According to the results of color ultrasound ,the cases were divided into three groups :IMT normal group(control group) ,thickening group and plaque group .According to the plaque echo characteristics ,the plaque group was redivided into two subgroups :stable plaque group and unstable plaque group .Serum Hcy and MIF levels and biochemical parameters were measured simultaneously in all cases .The differences of serum Hcy and MIF levels were compared between groups .The correlation coefficients among serum Hcy levels ,MIF levels and IMT were calculated .Results The serum Hcy and MIF levels in the control group ,thickening group and plaque group were increased in turn ,the difference among groups was statisti-cally significant(P<0 .01);which in the unstable plaque group was significantly higher than that in the stable plaque group ,the difference between them was statistically significant (P<0 .01) .The positive correlation was found among serum Hcy levels ,serum MIF levels and IMT (r=0 .584 ,0 .562 ,0 .607 ,P<0 .01) .Conclusion The serum Hcy and MIF levels are closely related with the carotid atherosclerosis degree and the plaque stability ;the serum Hcy and MIF levels are positively correlated with the carotid arte-rial IMT ;serum Hcy is positively correlated with the MIF level ,Hcy may cause the carotid atherosclerosis formation via MIF .

Objective:To investigate the expression of CD56 antigen in leukemia cells of the patients with acute myeloid leukemia (AML)and its relationship with the prognosis of AML, and to clarify the role of CD5 6 antigen expression in predicting the prognosis of the AML patients.Methods:171 AML (non-M3)patients aged from 14 to 60 years old,who received a IA Regimen as the first time inducing chemotherapy were chosen.Flow cytometric analysis was used to evaluate the CD56 expression in leukemia cells.COX proportional regression analysis was used to select the prognostic factors,and bivariable analysis was used to study the relationship between the positive rate of CD56 and overall survival (OS).The CD56+ group (n=52),including CD56≥50% expression group (n=39) and CD56<50% expression group (n=13),and CD56- group (n=119)were identified by the expression of CD56 antigen.The complete remission rate (CRR), the relapse rate, the median OS, the median disease-free survival (DFS)and the survival rate of patients were compared.Results:The medium OS of the patients in CD56+ group (14.2 months)was shorter than that in CD56- group (39.4 months)(P<0.05).Moreover,the medium OS in CD56≥50% group was shorter than that in CD56<50% group (11.7 months vs 20.3 months,P<0.05).The 1-year and 2-year survival rates of the patients in CD56+ group (61.5%,46.2%)were lower than those in CD56-group (75.6%,63.9%)(P<0.05).The 1-year survival rate had no significant difference between CD56≥50%group and CD56<50% group (53.8%vs 84.6%,P>0.05),while the 2-year survival rate in CD56≥50% group was lower than that in CD56<50% group (41.0%vs 61.5%,P<0.05).There were no significant differences of the CRR between CD56+ group (76.9%)and CD56- group (68.9%)as well as CD56≥50% group (58.9%)and CD56<50% group (63.5%)(P>0.05).The relapse rate and first year relapse rate of patients in CD56+ group (64.3% and 37.5%)were significantly higher than those in CD56- group (34.3% and 17.9% )(P<0.05). However,there were no significant differences of the relapse rate and first year relapse rate between CD56≥50%group (75.0% and 42.9%)and CD56<50% group (37.5% and 16.7%)(P>0.05).The DFS in CD56+ group was shorter than that in CD56- group (P<0.05).The same DFS result was also found between CD56≥50%group and CD56<50% group (P<0.05).Conclusion:The expression of CD56 antigen in leukemia cells predicts a bad prognosis in the AML patients,and the higher expression of CD56 indicates the worse prognosis.

To explore the new method of discriminating Astragali Radix and Hedysari Radix by using PCR amplification of specific alleles, 30 samples of the different Astragali Radix materials and 28 samples of Hedysari Radix were collected. The total DNA of all samples were extracted, trnL-trnF sequence from Astragali Radix and Hedysari Radix was amplified by PCR and sequenced unidirectionally. These sequences were aligned by using Clustul W. Primer was designed and the PCR reaction systems including annealing temperature, dNTP, etc were optimized. All samples were amplified by PCR with specific primer, DNA from Astragali Radix would be amplified 136 bp, whereas PCR products from all of Hedysari Radix were 323 bp. This method can detect 10% of intentional Hedysari Radix DNA into Astragali Radix. PCR amplification of alleles can be used to identify Astragali Radix and Hedysari Radix successfully and is an efficient molecular marker for authentication of Astragali Radix and Hedysari Radix.
Alleles ; Astragalus Plant ; classification ; genetics ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Polymerase Chain Reaction

OBJECTIVETo study the feasibility and authenticity of repairing alveolar defects in alveolar cleft patients with osteoinduction active material (OAM) in clinic.

METHODSTwenty-seven cases of alveolar defect chosen from clinic were divided into two groups (test group and control group). For test group (12 cases), OAM was transplanted to repair the alveolar cleft. For control group (15 cases), autogenous ilium cancellous bone were transplanted into the defect region to repair alveolar cleft. At 6 months after operation, CT and three-dimensional reconstruction were used to observe alveolar appearance, and the effect and clinical success rate of recover alveolar cleft by using different repair material were compared.

RESULTSIn the 27 cases, all the maxillary continuity was restored except two of test group and two of control group. There was no significant difference between test group and control group regarding the clinical success rate of the alveolar cleft repair (P = 1.000).

CONCLUSIONOAM was used to repair the alveolar cleft that can result in new bone formations and the burgeon of canines from the bone grafted areas. There is no significant difference between OAM and autogenous ilium cancellous bone regarding the effect of the alveolar cleft repair.

Alveolar Process ; pathology ; surgery ; Biocompatible Materials ; therapeutic use ; Bone Regeneration ; Bone Transplantation ; Cleft Palate ; surgery ; Humans ; Ilium ; transplantation

BACKGROUNDFractalkine is an important chemokine mediating local monocyte accumulation and inflammatory reactions in the vascular wall. Aspirin inhibits inflammatory cytokine expression closely related to atherosclerosis through the way independent of platelet and cyclooxygenase (COX). There has been no report about the effect of aspirin on fractalkine expression. We aimed to determine the fractalkine expression in human umbilical vein endothelial cell (HUVEC) stimulated by tumor necrosis factor (TNF)-alpha and the effect of aspirin intervention.

METHODSSix of 8 HUVEC groups received either different concentrations of aspirin (0.02, 0.2, 1.0, 5.0 mmol/L) or 40 micromol/L pyrrolidinecarbodithioc acid (PDTC) or 0.5 micromol/L NS-398. The other two groups were negative control and positive control (TNF-alpha-stimulated). After being incubated for 24 hours, cells of the 8 groups except the negative control one were stimulated with TNF-alpha (4 ng/ml) for another 24 hours. After that, the cells were collected for RNA isolation and protein extraction.

RESULTSBoth mRNA and protein expressions of fractalkine in HUVEC were upregulated by 4 ng/ml TNF-alpha stimulation. Aspirin inhibited fractalkine expression in a dose-dependent manner at mRNA and protein levels. Nuclear factor-kappa B inhibitor, PDTC, effectively decreased the fractalkine expression. Fractalkine expression was not influenced by COX-2 selective inhibitor NS-398. COX-1 protein expression was not changed by either TNF-alpha stimulation or aspirin, PDTC, NS-398 intervention. Both mRNA and protein expression of COX-2 in HUVEC were upregulated by 4 ng/ml TNF-alpha stimulation. Aspirin decreased COX-2 expression in a dose-dependent manner at mRNA and protein levels.

CONCLUSIONSTNF-alpha-stimulated fractalkine expression is suppressed by aspirin in a dose-dependent manner through the nuclear factor-kappa B p65 pathway.

Aspirin ; pharmacology ; Blotting, Western ; Cell Line ; Chemokine CX3CL1 ; genetics ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; pharmacology ; Umbilical Veins ; cytology

OBJECTIVETo study the effects of calponin-1 expression inhibition on the proliferation , invasiveness, apoptosis and cytoskeleton of uterine smooth muscle cells, and explore the molecular mechanism of calponin-1 in the uterine smooth muscle cells for labor onset.

METHODSsiRNA-calponin-1 adenovirus plasmid was constructed and transfected into primarily cultured uterine smooth muscle cells. The proliferation, invasiveness and apoptosis of the cells were determined by MTT assay, matrigel invasion assays and flow cytometry, respectively. Rhodamine-Phalloidin was used for labeling filamentous actin (F-actin), and the morphology and the distribution of F-actin was observed under fluorescence microscopy and analyzed quantitatively.

RESULTSThe motor ability of uterine smooth muscle cells decreased significantly after transfection with siRNA-calponin-1 adenovirus plasmid (P<0.05). The transfected cells showed thinner, loosened and irregular F-actin microfibers, and the cells in the empty vector and blank control groups showed thicker and longer F-actin microfibers.

CONCLUSIONInhibition of calponin-1 expression can inhibit uterine smooth muscle cell migration and cause the morphological change and rearrangement of F-actin without affecting its proliferation and apoptosis in vitro, suggesting that the morphological change and rearrangement of F-actin of uterine smooth muscle cell may be one of the important mechanisms in the labor onset.

Apoptosis ; Calcium-Binding Proteins ; genetics ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Female ; Gene Silencing ; Humans ; Microfilament Proteins ; genetics ; Myocytes, Smooth Muscle ; cytology ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Uterus ; cytology ; metabolism