Objective To investigate the effect of phenylhexyle isothiocyanate (PHI) on demethylation and activation of transcription gene p15 in acute leukemia cell line Molt-4. Methods DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Moh-4 cells were treated with PHI. P15 mRNA was measured by RT-PCR. Pl5 protein was detected by Western blotting. Results Hypermethylation of gene pl5 was apparently attenuated and activation of transcription p15 gene was de novo after 5 days exposure to PHI. PHI enhanced both the expression of p15 mRNA and p15 protein in a concentration-dependent manner. The ratio of the gray scale of p15 mRNA strap was 0.17±0.12 in control, 0.29±0.14 in PHI 10 μmol/L, 0.55±0.07 in PHI 20 μmol/L, 0.93±0.13 in PHI 40 μmol/L. Conclusion PHI could active demethylation and transcription of gene p15.

OBJECTIVE To investigate antibiotic resistance and the prevalence of serotype of Streptococcus pneumoniae in Wuhan.METHODS Totally 152 strains of S.pneumoniae were collected to test the MICs of various antibiotics by agar dilution method according to the approved standard of NCCLS.Serotyping of S.pneumoniae was performed by using quelling reaction.RESULTS Among 152 strains of S.pneumoniae,65(42.76%) strains were resistant to penicillin(MIC≥0.12mg/L).94.08%,50.66%,41.45% and 11.18% of S.pneumoniae were resistant against the first(cefalexin),second(cefaclor) and third(cefaxime and ceftriaxone) generation of cephalosporins respectively.The resistance rates to other antibiotic agents,such as erythromycin,tetracycline,trimethoprim/sulfamethoxazole and chloramphenicol,were 84.21%,88.82%,89.47% and 18.42%,respectively.Strains that were resistant to levofloxacin and moxifloxacin were found both for 1.32%.Twenty serotypes were involved in 152 strains. The prevalent serotypes were 19(25.66%),23(19.08%),6(13.82%),15(7.24%)and 14(4.61%).Eight strains were remained for unable to serotype.All penicillin-resistant S.pneumoniae was included in serotypes 6,19 and 23.CONCLUSIONS The antibiotic resistance of S.pneumoniae is serious in Wuhan.Most of them are multi-resistant strains.Except for fluoroquinolones,ceftriaxone and chloramphenicol, most antibiotic agents have lost there activities against S.pneumoniae.The prevalent serotypes,especially of the multi-resistant strains,were 19,23 and 6.Pneumococcal polyvalent vaccine can well cover these serotypes.

Objective To retrospectively analyze the surgical methods and efficacy in 70 cases of type A aortic dissection patients over 65 years old.Methods From January 2005 to May 2012,70 type A aortic dissection patients over 65 years old received surgical treatment.Among them,there were 47 males aged 65 to 78 years old with mean 71,23 females,aged 65 to 72 years old with mean 68.55 cases were acute onset,while 15 cases were chronically onset.Different surgical methods were selected depend on patients' situations.We followed up all patients after discharged from hospital to continue to observe their health situation and evaluate the therapeutic effects.Results After surgery,eight patients died in the hospital,62 patients were recovered and discharged from the hospital.The mortality rate is 11.4％.During the follow up period from 3 to 72 months,there were no dead,aneurysm rupture and others severe complications.9 cases received endovascular graft exclusion within 6 months after discharged from hospital.The survival patients were satisfactory healed with their daily living activity resumed.Conclusion For over 65 years old patients with aortic dissection,the accurate and rapid selection of surgical method could improve the survival rate and the quality of life with a lower occurrence rate of complications.

A rapid and simple ultra performance liquid chromatographic method for determination the specific migration of bisphenol A diglycidyl ether ( BADGE ) , bisphenol F diglycidyl ether ( BFDGE ) and their derivatives in food simulants was developed. Water, 3% acetic acid, 10% ethanol and sunflower oil were used as food simulants to simulate the specific migration of bisphenol diglycidyl ethers from interior coating of food cans after 10 days storage at 60℃. After the migration period, the aqueous food simulants were directly measured without any further purification, while the sunflower oil simulant was extracted by acetonitrile followed by cleaning up using solid phase extraction. Among the migration process, BADGE and bisphenol A (3-chloro-2-hydroxypropyl)glycidyl ether (BADGE·HCl) migrated into aqueous simulants were hydrolysed into bisphenol A bis ( 2, 3-dihydroxypropyl ) ether ( BADGE · 2H2 O ) and bisphenol A ( 3-chloro-2-hydroxypropyl) (2,3-dihydroxypropyl) ether (BADGE·H2O·HCl), respectively. However, BADGE and BADGE·HCl migrated into sunflower oil were not hydrolysed. The calibrating curves showed a good linearity from 0. 05 to 10 mg/L for all the 9 target compounds. The detection limits of the method for aqueous food simulants and sunflower oil stimulant were 5μg/L and 20μg/kg, respectively. The method was applied to the determination of bisphenol diglycidyl ethers migrated from 10 kinds of food cans which were intended to contact with food. The results indicate that BADGE and its derivatives were detected at 5 of the cans, and the specific migration of BADGE(or BADGE·2H2O)and BADGE·HCl(or BADGE·H2O·HCl)in 1 cans even exceed the responding limitation regulated in EC/1895/2005 .

Objective To characterized the Salmonella enterica serotype typhimurium isolates recovered during 2002 to 2005 from outpatients in Tongji Hospital, Wuhan China. Methods The 36 isolates from Tongji Hospital were characterized by antimicrobial-susceptibility testing and screened for class Ⅰ integrons, beta-lactamase genes, qnr, aac(6')-Ib-cr and mutations in the quinolone resistance determining regions (QRDRs) by PCR and DNA sequence analysis. All isolates were also characterized by pulsed-fieldgel electrophoresis (PFGE) to determine the genetic relateness among these isolates. Results All isolates displayed multidrug resistance and most of them harbored class Ⅰ integrons. Ciprofloxacin-resistant isolates showed significant difference compared with ciprofloxacin-susceptible isolates after PFGE analysis. All 31 ciprofloxacin resistant isolates carried at least three mutations in the QRDR of GyrA and ParC. Three ciprofloxacin resistant isolates had accumulated additional mutation in ParE. Five isolates harboring the OXA-30. Enzyme showed intermediate resistant to eefepime. Conclusions Fluoroquinolone-resistant Salmonella typhimurium isolates were widely distributed among the outpatients in Wuhan and the resistant isolates accumulated multiple antimicrobial resistant mechanisms and showed unique genetic profiles. The state and local health authority must remain vigilant for the emergence of Salmonella typhimurium resistant to both third generation cephalosporins and fluoroquinolonos.

Objective To retrospectively evaluate the role of consolidative repeat radiofrequency ablation (CRRFA) based on safety margin (SM) analyses in local tumor control for alpha-fetoprotein (AFP) negative hepatocellular carcinoma (HCC) patients who had been shown to have radiological complete ablation (CA) with radiofrequency ablation (RFA). Methods From July 2002 to July 2009,152 AFP negative HCC patients who were shown to have radiological CA with RFA therapy were retrospectively analyzed. Among them, 110 patients had a SM of less than 1 cm and the other 42 patients had a SM of 1cm or more. Among 110 patients with SM less than 1 cm, fifty nine patients accepted CRRFA within 6 months after the first RFA and 51 did not. From these patients, a narrow SM-CRRFA group (n=41) and a narrow SM-single RFA group (n=37) were enrolled respectively. The wide SM-single RFA group (n= 30) was enrolled from the 42 patients with a SM of 1 cm or more.The LTP (local tumor progression)-free survival rate of the 3 groups were compared with a log-rank test. Results One-, two-, three-, four-, and five-year LTP-free survival rates respectively were 97. 1%, 90.9%, 69.6%, 47.2%, and 33. 0% in the narrow SM-CRRFA patients. 85.9%, 66. 5%,43.5%, 15.8%, and 0. 0%, in the narrow SM-single RFA patients, and were 92.7%, 83.7%,59.3%, 36. 9%, and 9.2% in the wide SM-single RFA patients. There were statistically significant differences (χ2 = 14. 789, P= 0. 001) between the groups. Conclusions An ablation zone with an SM of 1 cm or greater was the most important factor for local control of AFP negative HCC ranging from 3 to 5 cm in diameter. For these patients with a SM of less than 1 cm, CRRFA improved the overall local control outcomes.

Objective To investigate the different effect of exogenous leptin on estrogen receptor α,β mRNA in human breast tumor tissue in nude mice xenograft models. Methods We made nude mice xenograft models of MCF-7 human breast cancer cells cultured in vitro, then divided them into experimental group of]eptin( n = 30)and control group of normal saline( n = 30)randomly. The models of experimental group were injected subcutaneously the recombinant human leptin for 15 consecutive days, the models of control group were injected subcutaneously the same dose of normal saline. A real- time quantitative RT- PCR assay was developed to quantify the expression of estrogen receptor α, β mRNA in tumor tissue, using the relative quantitative analysis. Results The leptin-intervened nude mice xenograft models were safely established. The relative quantitation of estrogen receptor α mRNA was significantly higher in the leptin group than in the normal saline group ( P ＜ 0.01 ), the relative quantitation of estrogen receptor β mRNA was significantly lower in the leptin group than in the normal saline group ( P ＜ 0. 01 ). Conclusion The nude mice xenograft models can be safely intervened with human leptin by subcutaneous injection around tumor.Estrogen receptor is one of the targets of leptin in the progress of breast cancer. Exogenous human leptin can up- regulate the expression of estrogen receptor α and down- regulate the expression of the estrogen receptor βin nude mice xenograft models of human breast tumor.

Objective To investigate phenyhexyle isothiocyanate (PHI) modulating histone acetylation and inhibiting Akt signaling pathway in prostate cancer cell line PC3 in vitro. Methods Apoptotic cells were measured by TUNEL assay. Histone acetylated H3, H4 and the Akt protein signaling pathway (Akt, p-Akt, mTOR, p-mTOR, p70S6K and p-p70S6K) were detected by Western blot. Results Apoptotic cells increased after exposure to PHI with concentration dependent. PHI significantly induced an accumulation of histone acetylated H3, H4. The change of Akt, mTOR, p70S6K proteins was not observed. Phosphorylation of Akt (p- Akt), mTOR (p-mTOR) and p70S6K (p-p70S6K) decreased after exposure to PHI for 7 h. Conclusions PHI can induce histone acetylation H3, H4 accumulation. PHI inhibits Akt signaling pathway resulting cell apoptosis. It might be a new anticancer agent.

Objective To study on plasmid-mediated quinolone-resistant in Escherichia coli strains isolated from fecal samples of chicken and swine from the nine farms around our country.Methods Antimi-crobial susceptibility testing was carried out by broth microdilution testing,gyrA,gyrB,parC,qnr and aac (6')- Ⅰ b-cr were examined by PCR,and the products were sequenced.Conjugation experiment was carried out to proved that the plasmid-mediated quinolone resistance was transferable.Results In the total 818 animal isolates,qnr and aac genes were detected in 38 (4.6%) and 75 (9.2%) strains.The qnrA,qnrB,and qnrS genes were detected in 1 (0.1%),9 (1.1%) and 28 (3.4%) of the isolates.All isolates were negative for qnrC,qnrD genes.Conclusion There is a close relationship between high level quinolone resistance and plasmid-mediated quinolone resistance.The results of the current study highlight food-producing animals as a potential reservoir of antimicrobial-resistant bacteria and clinically important resistance genes.More attention should be paid to the surveillance of such strains.

Objective To characterize the roles of different quinolone resistance mechanisms in quinolone-resistant Escherichia coli isolates,including different topoisomerase point mutations,efflux pumps and outer membrane proteins.Methotis Through homologous gene recombination methods,different quinolone-resistant mechanisms of E. coli mutants were constructed and the susceptibility changes of these mutants to different antimicrobials were measured.Resuits Efflux pumps AcrAB and outer membrane protein TolC played different roles in different E. coli isolates.Compared with other mechanisms,the mutations in topoisomerases played a dominant role in quinolone resistance.Only the mutations jn parC had no effect on quinolone resistance,which further confirmed parC was the secondary target of quinolones in E.coli.Fluoroquinolone susceptible E.coli would automatically become highly resistant to quinolones after acquiring the point mutations in both gyrA(S83L,D87N)and parC(S80I,A108V),but not requiring the over-expres-sion of efflux.Conclusion The mutations in topoisomerases play a dominant role in E.coli quinolone resistance,and the mutations in both gyrA and parC are required.