Objective To investigate the effect of MDM2-p53 feedback loop on the sensitivity to cis- platin in ovarian cancer xenograft in vivo and explore its mechanism.Methods Human ovarian cancer cell line A2780 with wild type p53 was inoculated subcutaneously into the back of nude mice.When tumor nod- ules could be observed after 7 days of inoculation,the mice were randomly divided into 5 groups with each of 6 mice.Treatment group received an intratumoral injection of a combination of pCMV-MDM2-Lipofectamine and cisplatin.Control groupⅠwas injected with cisplatin,control groupⅡwas injected with pCMV-MDM2- Lipofectamine,control groupⅢwas injected with empty pCMV-Lipofectamine,control groupⅣwas injected with RPMI1640.General conditions and tumor growth rate were observed.The volume and the weight of the tumor were compared among these five groups.The expression of MDM2 and p53 in these tumors were de- tected by immunohistochemistry and Western blot.The changes of cell cycles in these tumors were examined by using flow cytometry assay.Results Tumor volume in treatment group[(0.321?0.086) cm~3]was significant- ly smaller than control groupⅠandⅡ[(1.832?0.165) cm~3 and (3.251?0.179) cm~3,respectively)].The weight of the treatment group [(0.513?0.089) g]was also significantly lower than control groupⅠandⅡ[(1.412?0.134) g,and (2.665?0.153) g,respectively)].There was a significant difference between control groupⅠandⅡ, but no difference between the other three control groups.Obvious MDM2 protein expression and pronounced S-phase arrest were observed in treatment group.However,control groupⅠwas with intact wild type p53,and arrested primarily in G_2/M phase.Conclusion MDM2 overexpression can increase cisplatin cytotoxicity on experimental ovarian cancer through inhibition of p53 expression and the loss of G_1 check point of cell cycle.

Objective To discuss the active role of health education on control of diabetes mdlitus and prevention of its complications.Methods 96 cases of hospitalized patients with diabetes mellitus were randomly divided into the control group and the observation group with 48 patients in each group from June 2009 to June 2010.The control group received conventional diabetes treatment and distribution of health education brochures,based on this,the observation group was given health education,including cognitive,nutrition,behavioral intervention.The general prevention condition of diabetes and its complications as well as the education effect were compared before the education and six months after education.Results The fasting blood glucose,2-hour postprandial blood glucose,glycosylated hemoglobin,body mass index and incidence of complications were greatly improved in the observation group than those of the control group.Conclusions Strengthening health education can improve self-care ability of patients,effectively improve the overall control level of diabetes,reduce acute and chronic complications,disability,death rate.

0.05).Our data also showed no significant association between the genotypes and the severity of the disease.One-way ANOVA showed that BDNF genotype had no association to the age of onset for developing AD.Conclusions Our results indicate that Va166Met SNP in BDNF gene is not associated with AD.

By using a cell-based high throughput screening model for the CLA-1 up-regulator, Streptomyces 203909 was found to produce up-regulator of CLA-1. A novel trichostatin analogue was isolated from the rice fermentation of Streptomyces sp. CPCC 203909by a combination of various chromatographic techniques including column chromatography (CC) over silica gel, flash C18 CC, and reversed-phase HPLC. Its structure was identified as (-)-(R,2E,4Z)-7-[(4'-dimethylamino) phenyl]-4,6-dimethyl-7-oxohepta-2,4-dienoyl-L-glutamine (1) by the spectroscopic and chemical methods, and combination with the CD spectroscopy and Marfey's method. In the prelimi- nary assays, Compound 1 showed cytotoxicity against human embryonic kidney 293 cell line with IC50 value 35.3 [µmol · L(-1).
Cell Survival ; drug effects ; Fermentation ; Hep G2 Cells ; Humans ; Hydroxamic Acids ; chemistry ; isolation & purification ; metabolism ; pharmacology ; Molecular Structure ; Streptomyces ; chemistry ; metabolism

BACKGROUNDCandida albicans is the most frequently seen opportunistic human fungal pathogen. Terbinafine is an allylamine antifungal agent that has been proven to have high clinical efficacy in the therapy of fungal infections, the mechanism of action of terbinafine involves the specific inhibition of fungal squalene epoxidase, resulting in ergosterol deficiency and accumulation of intracellular squalene. We used cDNA microarray analysis technology to monitor global expression profile changes of Candida albicans genes in response to terbinafine treatment, and we anticipated a panoramic view of the responses of Candida albicans cells to the representatives of allylamine antifungal agents at the molecular level in an effort to identify drug class-specific and mechanism-independent changes in gene expression.

METHODSCandida albicans strain ATCC 90028 was exposed to either medium alone or terbinafine at a concentration equivalent to the 1/2 minimal inhibitory concentrations (MICs, 4 mg/L) for 90 minutes. RNA was isolated and gene expression profiles were compared to identify the changes in the gene expression profile using a cDNA microarray analysis. Differential expression of 10 select genes detected by cDNA microarray analysis was confirmed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSA total of 222 genes were found to be responsive to terbinafine, including 121 up-regulated genes and 101 down-regulated genes. These included genes encoding membrane transport proteins belonging to the members of the ATP-binding cassette (ABC) or major facilitator superfamily (MFS; CDR1, AGP2, GAP6, PHO84, HOL3, FCY23, VCX1), genes involved in stress response and detoxification (CDR1, AGP2, HOL3), and gene involved in the ergosterol biosynthesis pathway (ERG12). The results of semi-quantitative RT-PCR were consistent with that of the cDNA microarray analysis.

CONCLUSIONSThe up-regulation of the gene encoding the multidrug resistance efflux pump CDR1 may contribute to the terbinafine resistance in Candida albicans. However, the precise roles of other affected genes remain unclear, further studies of these genes and their respective products that play roles in the context of antifungal resistance are warranted.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; genetics ; Ergosterol ; biosynthesis ; Fungal Proteins ; genetics ; Gene Expression Profiling ; Genome, Fungal ; Membrane Transport Proteins ; genetics ; Naphthalenes ; pharmacology ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo study the role of RpoE and RpoS on the influence of the metabolism and growth of bacterial under hyperosmotic stress.

METHODSThe rpoS/rpoE double deletion mutant of Salmonella enterica serovar typhi (S. typhi) was prepared by homologous recombination through the suicide plasmid mediated. The recombination was visualized by PCR. Growth curves were drawn by using photometric value A600 as the ordinate and cultivation time as abscissa. The survival abilities of bacterial were compared under hyperosmotic stress. Statistical differences of early logarithmic growth stage (4 h) and laters logarithmic growth stage (12 h) were analyzed by one-way ANOVA. The expression difference of metabolism related genes of wild-type and mutant strains of S. Typhi incubated under hyperosmotic stress were investigated by Salmonella genomic DNA microarray. Real-time quantitative PCR (qRT-PCR) was performed to validate the results of microarray assay in some selected genes.

RESULTSThe rpoS/rpoE double deletion mutant of S. Typhi was successfully generated. The analysis of growth curve showed that the 4-hour and 12-hour A600 values were separately 0.503 ± 0.018 and 2.060 ± 0.112 in rpoS deletion mutant strains, 0.293 ± 0.053 and 1.933 ± 0.115 in rpoE deletion mutant strains, and 0.051 ± 0.007 and 0.963 ± 0.111 in rpoS/rpoE double deletion mutant strains; all of which were lower than the values of wild-type strains, who were 0.725 ± 0.097 and 2.496 ± 0.171, respectively. The difference were statistically significant (P < 0.05). The genomic DNA microarray revealed that 42 genes relevant with bacterial metabolism were influenced by RpoE and RpoS. Results of qRT-PCR showed that the expression values of rpsE, rbsK, nusG and etuB in rpoS deletion mutant strains were (1.86 ± 0.14)×10(6), (1.37 ± 0.11)×10(6), (2.72 ± 0.58)×10(6) and (8.27 ± 1.01)×10(6) copies/µg, respectively; while those in rpoE deletion mutant strains were (2.19 ± 0.17)×10(6), (1.51 ± 0.12)×10(6), (2.73 ± 0.57)×10(6) and (9.63 ± 1.42)×10(6) copies/µg, respectively. Compared with the values in wild-type strains, which were separately (1.94 ± 0.10)×10(6), (1.52 ± 0.11)×10(6), (2.39 ± 0.52)×10(6) and (10.83 ± 1.52)×10(6) copies/µg, the differences was not statistical significance (P > 0.05). However, compared with the values in rpoS/rpoE double mutant strains, which were separately (5.64 ± 0.59)×10(6), (4.17 ± 0.40)×10(6), (9.44 ± 1.22)×10(6) and (2.95 ± 0.88)×10(6) copies/µg, the difference was significant (P < 0.05).

CONCLUSIONRpoE and RpoS could influence the expression of lots of metabolism genes. Together, they regulated the metabolism and growth of S. Typhi under hyperosmotic stress.

Bacterial Proteins ; genetics ; Gene Deletion ; Osmosis ; Salmonella typhi ; genetics ; growth & development ; metabolism ; Sigma Factor ; genetics ; Stress, Physiological

OBJECTIVETo design DNA microarray and investigate the molecular anti-tumor mechanism of herbs of traditional Chinese medicine.

METHODcDNA microarrays consisting of 56 probes representing 24 human cell cycle genes were constructed, Four anti-hepatocarcinoma herbs including Radix Linderae, Hebra Artemisiae Annuae, Radix Amebiae, Radix Astragli, were chosen. Effects of herbs on SMMC-7721 cell cycle were observed by flow cytometry assay. Effects of herbs on cell cycle gene expression in SMMC-7721 cells were analyzed by comparing hybridization of Dig-Labeled cDNAs from herb-treated cells and cDNAs from untreated cells.

RESULTExpressions of cell cycle geneswere changed in different degrees after herbs treated. Some genes were down-regulated and some genes were up-regulated. The changes in gene expression agreed with the results of flow cytometry assay.

CONCLUSIONThe results suggest that these herbs may have effects on cell cycle and DNA damage checkpoint genes which may be the mechanism of the herbs, and DNA microarray can be used to investigate the biological function of extracts of traditional Chinese medicine.

Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Artemisia ; chemistry ; Astragalus membranaceus ; chemistry ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Cyclin-Dependent Kinases ; genetics ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gene Amplification ; Gene Expression Profiling ; Genes, cdc ; drug effects ; Humans ; Lindera ; chemistry ; Lithospermum ; chemistry ; Liver Neoplasms ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis ; methods ; Plants, Medicinal ; chemistry ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; cdc25 Phosphatases ; genetics ; metabolism

OBJECTIVETo investigate the effect of homoharringtonine (HHT) on leukemic stem-like cells (LSC) in human acute myeloid leukemia (AML) cell lines.

METHODSThe phenotypes of AML cell lines U937,Kasumi-1,and KG-1 cells were analyzed by flow cytometry (FACS). The effect of HHT on leukemia stem-like cells with immunophenotype of CD34(+)CD38(-)CD96(+) was detected with FACS. Cell growth was measured by MTT assay. Activation of Caspase pathway and expression of apoptosis-related regulator proteins were examined by Western blotting.

RESULTSFACS demonstrated that the 69% of KG-1 cells expressed LSC phenotype CD34(+)CD38(-)CD96(+), while 26.7% on Kasumi-1 cells expressed this marker. In contrast,U937 cells showed CD96 negative. HHT significantly inhibited cell growth of KG-1 cells with an IC(50) of 16.9 ng/ml at 48 h. The ratio of CD34(+)CD38(-)CD96(+) cells decreased from 63.6% to 17.1% after HHT treatment. Enhanced apoptosis was demonstrated in HHT group evidenced by strong activation of Caspase-9,Caspase-3 and PARP.HHT treatment resulted in down-regulation of expression of anti-apoptotic protein BCL-2 and phosphorylated-Akt.

CONCLUSIONHHT can effectively kill the leukemic stem-like cells in human AML cell line KG1 by inhibiting cell growth and inducing apoptosis which is associated with activation of Caspase pathway and down-regulation of anti-apoptotic proteins and phosphorylated-Akt.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Harringtonines ; pharmacology ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Neoplastic Stem Cells ; drug effects ; metabolism ; pathology

BACKGROUND: The immediate implantation in the anterior maxillary region is in a high risk of aesthetic complications. OBJECTIVE: To assess the prognosis of the labial bone plate after anterior maxillary repair with immediate implant combined with immediate restoration. METHODS: Thirty-two patients with single failed tooth in the anterior maxillary region were subjected to implantation of ZIMMER implants immediately after minimally invasive extraction. Good primary stability was achieved and immediate restoration was carried out. Final restoration was finished after 6-12 months of osteosynthesis and gingival shaping. The loading situation of the labial bone plate was recorded at 6 months post operation. RESULTS AND CONCLUSION: Final restoration was finished with normal loading in all the patients. No bleeding and swelling of the gingiva was recorded. The horizontal absorption of the labial bone plate at the upper margin, 5 mm and 10 mm below the upper margin was (-2.12±0.05), (-1.54±0.04), and (-1.01±0.06) mm, respectively. Therefore, absorption of the labial bone plate with varying degrees exists after anterior maxillary repair with immediate implant combined with immediate restoration.

The development of a RF ablation therapeutic instrument based on improved PID algorithm is introduced here. It is based on the theory of radio frequency local destruction. By adopting the improved PID temperature control algorithm, the problem of the temperature control precision reduction due to tumor tissue characteristic changing by heating has been solved, thus ensuring homogeneous and smooth radio frequency heating to tumor foci. Experiments demonstrate that the new algorithm has strong adaptability and anti-disturbance capability, the equipment works stably and reliably, and can control therapeutic temperature precisely (+/- 2 degrees C), which indicates a good clinical application value.
Algorithms ; Calorimetry ; instrumentation ; methods ; Catheter Ablation ; instrumentation ; methods ; Electrodes ; Equipment Design ; Humans ; Hyperthermia, Induced ; instrumentation ; methods ; Information Storage and Retrieval ; Neoplasms ; therapy ; Software ; Temperature