Objective: To investigate the effect of cell division cycle 42 (Cdc 42) gene on the formation of filopodia suppressed by non-steroidal anti-inflammatory drugs (NSAIDs), and to explore the probable signal transduction pathway involving Cdc 42 in the inhibition effects on migration and invasion abilities of breast cancer cells induced by NSAIDs. Methods: MCF-7 cells were divided into four groups: blank control group, NS-398 (100 μmol/L)-treated group, cyclooxygenase-2 (Cox-2) small interfering RNA (siRNA)-transfected group and the negative siRNA-transfected group. The expressions of Cox-2 and Cdc42 mRNAs were detected by real-time fluorogentic quantitative PCR(RFQ-PCR), and the expressions of Cox-2 and Cdc42 proteins were measured by Western blotting. Actin-tracker Green fluorescent probe was used to examined the morphology of filopodias in MCF-7 cells. The invasion ability of MCF-7 cells was detected by using the Martrigel-coated Transwell. Results: The filopodias in MCF-7 cells disappeared in the NS-398-treated group, and the invasion ability of MCF-7 cells was also significantly decreased in this group compared with that in the blank control group (P<0.05). There was no difference in the expression level of Cox-2 mRNA or protein between the blank control group and the NS-398-treated group (P>0.05), and the difference was also not seen in the expression of Cdc42 mRNA or protein between these two groups (P>0.05). The active-Cdc42 level was significantly decreased in the NS-398-treated group compared with that in the blank control group (P<0.05). The filopodias in the Cox-2 siRNA-transfected group disappeared, and the invasion ability of MCF-7 cells transfected with Cox-2 siRNA was significantly decreased compared with that in the negative siRNA-transfected group (P<0.05). The active-Cdc42 level was also significantly decreased in MCF-7 cells transfected with Cox-2 siRNA compared with those transfected with negative siRNA (P<0.05). Conclusion: NSAIDs can suppress the invasion ability of breast cancer cells. This effect may be achieved by inhibiting the activity of Cox-2, decreasing the expression level of active-Cdc42, and suppressing the formation of filopodias.

Coronary Artery Disease ; pathology ; Humans ; Medicine, Chinese Traditional

Humans ; Male ; Middle Aged ; Pneumoconiosis ; diagnosis

Objective To identify the role of RNA-editing enzyme ADAR1 (adenosine deaminase acting on RNA) in EV71 infection and virus mutation.Methods RNAi technology was applied to establish ADAR1 knock-down stable cell lines.Then the cells were served to evaluate the role of ADAR1 in EV71 infection by MTT assay for detecting virus-induced cell viability,virus plaque assay for quantification of the virus titer and the cellular susceptibility to the virus,and Western blot for virus protein expressions.ADAR1-mediated RNA editing can result in the genetic A-G and T-C mutations.To further determine whether the effects of ADAR1 on EV71 infection were correlated with ADAR1-mediated EV71 RNA editing and therefore increased the viral mutations during the infection,the characteristics of EV71 mutation were analyzed based on the different full-length viral genomes from epidemic regions.The viral genome was also sequenced from the infected ADAR1 knock-down cells.Results After ADAR1 knock-down,the cell viability decreased quickly after the virus infection,and formed much more and larger sizes of plaques than the control cells.The virus capsid protein VP1 expressions and virus titer in the cells culture media were both increased in ADAR1 knockdown cells.Statistic analysis showed that A-G and T-C mutations were the major mutations of EV71,which were believed to be the hot sites for RNA-editing.However,the results of viral RNA genomic sequencing data indicated that ADAR1 did not edit EV71 genome directly.Conclusions ADAR1 was a restriction factor for controlling EV71.However,ADAR1 does not directly edit EV71 genome.

Objective To detect cytokeratin in routine pathology negative regional lymph nodes postoperatively in non-small cell lung carcinoma (NSCLC). To investigate the relationship of lymph node micrometastasis in P-TNM stages NSCLC and survival rates. Methods From Jan. 1996 to Dec. 2003, 107 paraffin-embedded specimens of T1-T4N0-N1M0 NSCLC patients were collected. Anti-cytokeratin(CK) an- tibody AE1/AE3 was applied to detect cytokeratin with Envision~(TM) method in routine pathological negative re- gion lymph nodes in NSCLC, and selected negative control, positive control and blank control. The pulmo- nary hilar lymph node micrometastasis was upward regulated with stage pCK-N1, mediastinal lymph node mi- crometastatsis was upward regulated with stage pCK-N2. The result applied to SPSS11.0 software to process. Results The CK positive rate was 29.9% in all the patients. The CK positive rate was 27% (21/78), 30% (7/23), 67% (4/6)in stage p-Ⅰ, p-Ⅱand p-Ⅲ, respectively. All these data showed the tendency by which detectable rate increased and was accompanied by disease progress. Comparing the annual survival rate and median survival time of the non-micrometastasis group with the mierometastasis group in two groups, the survival rate difference was statistically significant. Comparing the annual survival rate and median sur- vival time in pCK-ⅢA stage with p-Ⅰ-Ⅱstage, pCK-ⅢA stage annual survival rate and median survival time was significantly different (P=0.020). Similarly, comparing the survival rate in pCK-ⅡB stage with p-ⅠB stage, pCK-ⅡB stage survival rate was significantly different(P=0.059). Comparing the survival time of pCK-ⅢA stage with p-Ⅲstage, pCK-ⅡB stage, with p-ⅡB stage, euther survival time difference was statistically significant (P=0.838, 0.518). Conclusions The rate of positive cytokeratin increase is ac- companied by the disease progress in NSCLC. Positive cytokeratin has disadvantagious prognosis. It is showed that pCK-N1 may be equal to p-N1 and pCK-N2 which also may be equal to p-N2. Micrometastasis may affect the UICC staging currently in use.

Objective To observe the clinical behaviors of cervical intraepithelial neoplasia (CIN) for early diagnosis to improve the therapeutic efficacy Methods Eighty seven cases with different grades of CIN confirmed with colposcopy and pathological examination were reviewed for the clinical features and the results from colposcopy, pathology, testing for HPV (human papillomavirus) All patient were treated with various methods and followed up Results Most patients with CIN were in childbearing period and sought for medical advices because of increased leucorrhagia and/or bloody leucorrhea, postcoital bleeding for a comparatively long course Chronic cervicitis was often found in these patients Contact bleeding and HPV infection were common in patients with CINⅡand CIN Ⅲ Patients with CIN Ⅰ were regular followed up or treated with electrocautery, those with CIN Ⅱ were treated with conization or hysterectomy, and for those with CIN Ⅲ, hysterectomy was performed The outcomes of all patients were satisfactory after treatment Conclusion The most common sign for the patients with CIN is postcoital bleeding For correct diagnosis of CIN, colposcopy and together pathological examimotion is recommended High risk HPV detection is a good prognostic factor After treatment, the patients with different grades of CIN should be followed up regularly

OBJECTIVE To investigate the distribution and the resistance of pathogens from the children with VAP in PICU,and to analyze the reasons of antibiotics resistance of the pathogens.METHODS The sputum obtained from the children with final diagnosis of VAP in PICU was cultured and identified from Jan 2005 to Dec 2006.The resistance of the bacteria identified to antibiotics used frequently was determined by Kirby-Bauer method.RESULTS A total of 187 strains were isolated from sputum specimens,of which Gram-negative bacilli and Gram-positive cocci accounted for 76.5% and 23.5%,respectively.Acinetobacter baumannii(17.7%),Escherichia coli(16.0%),Pseudomonas aeruginosa(15.0%) and Klebsiella pneumoniae(13.9%) were the most frequently isolates of Gram-negative bacilli.Their resistant rates to ?-lactam antibiotics were high,especially the ESBLs-producing strains in E.coli and K.pneumoniae.The Staphylococcus epidermidis(5.9%),Staphylococcus aureus(4.8%) and Enterococcus faecalis(4.3%) were the most common strains of Gram-positive cocci.No vancomycin-resistanct strains were found,but resistance rates to ?-lactam antibiotics and other antibiotics were high in S.epidermidis and S.aureus.CONCLUSIONS The main strains cultured from the sputum specimens of children with VAP in PICU are Gram-negative bacilli with high resistance rates to antibiotics,especially the ESBLs producing bacilli to ?-lactam antibiotics.Staphylococcus are the main Gram-positive cocci.

Objective: To optimize the vacuum belt drying process of Shiwei Penan Granules. Methods: On the base of single factor experiments, feeding rate, heating temperature, and drying time were selected as the main factors, with composite score of moisture content of dry paste, rates of polydatin and paeoniflorin transfer as the response values, response surface methodology was used to optimize the vacuum belt drying process. Results: Optimal parameters were as follows: The feeding rate was 1.6 kg/h, heating temperatures was 105℃, drying time was 104 min; Under these conditions, moisture content of dry extract was 2.80% on average, the transfer rate of polydatin was 92.55%, and the transfer rate of paeoniflorin was 94.77% on average, the composite score was 95.35 on average, RSD=0.42%. Conclusion: The condition optimized by using response surface methodology is stable and reasonable, which could guide industrial production and keep high transfer rate of effective component.

OBJECTIVE: To evaluate the sub-acute oral effect of titanium dioxide (TiO₂) nanoparticles on the oxidation/antioxidation biomarkers and inflammatory cytokines in blood, liver, intestine, and colon in rats. METHODS: Twenty four 4-week-old clean-grade Sprague Dawley (SD) rats were randomly devided into 4 groups by body weight (n=6, control, low, middle, and high), in which the rats were orally exposed to TiO₂ nanoparticles at doses of 0, 2, 10 and 50 mg/kg body weight/day for 28 consecutive days separately. Food intake, body weight and abnormal behaviors during the experiment were recorded. The rats were euthanized on the 29th day. The blood was collected via abdominal aortic method and centrifuged to collect the serum. Tissues from liver, intestine and colon were collected and homogenated. Then enzyme-linked immunosorbent assay (ELISA) and microwell plate methods were used to detect oxidation/antioxidation biomarkers including superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px), total mercapto (T-SH), glutathione disulfide (GSSG), malomdialdehvde (MDA) and inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the serum, liver, intestine and colon in the rats. RESULTS: Compared with the control group, no significant differences in body weight, food intake and organ coefficients were observed in all the three groups after TiO₂ gavage. No significant changes in GSH, GSH-Px, T-SH, and IL-6 were observed. Compared with the control group, significant increase of SOD activity in serum in high dose group, signi-ficant increase of GSSG concentration in intestine in middle and high dose group and significant increase of MDA concentration in liver in low and high dose group were observed. Compared with the control group, a significant increase of TNF-α in liver in middle and high dose group was observed. CONCLUSION TiO₂ nanoparticle can increase antioxidant enzymes activities in blood, increase oxidative biomarkers in liver and intestine, increase inflammatory cytokines in liver in rats after a 28-day sub-acute orally administration. Among blood, liver, intestine, and colon, liver is most sensitive to the toxicity induced by TiO₂ nanoparticles, followed by intestine, blood, and colon in sequence.
Animals ; Antioxidants ; Biomarkers ; Cytokines ; Nanoparticles ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Titanium