Objective:To construct a expression vector of EGFR specific siRNA and transfect it into human ovarian cancer cell line SKOV3,then to evaluate the effect of RNA interference(RNAi) on the target gene expression and radiosensitivity of the cell line.Methods:EGFR sh-RNA was synthesized in vitro and transfected into SKOV3 cells with lipofectamine technique and then the positive clones were selected by G418.The positive RNA interference cell line,negative RNA interference cell 1ine and the normal control cell 1ine were included in the study.RT-PCR and Western Blot technique were performed to detect the inhibitory effect of RNAi on EGFR mRNA level and protein level,respectively.Three kinds of cell lines were exposed to different doses of radiation.Cell survival curve was drawn by means of MTT method.Results:The expression of EGFR gene was surpressed obviously in the specific sequence shRNA group,and EGFR mRNA and protein surpression rates were 77.5% and 76.1%,respectively;Sequence-specific shRNA-EGFR can significantly enhance the radiosensitivity of ovarian cancer cells,and significant statistical differences existed among the three groups(P

Objective To establish a method for detecting the EGFR gene mutations by the real-time fluorescence quantification PCR combined with Sanger sequencing and to preliminarily explore its clinical application value .Methods With EGFR gene hotspot mutations region exon 19 and 21 as the research locus ,the specific amplification and the sequencing primer were designed ,the known wild-type and mutant samples were utilized to construct the corresponding plasmid as the standard substance by the TA clone technique .Then the EGFR gene mutation detection method by the real-time fluorescence quantification PCR combined with Sanger sequencing was established and the methodological and the application evaluation were performed .Results The wild-type and mutant standard plasmids of the EGFR gene exon 19 and 21 were constructed successfully .The EGFR gene mutations detection method of the real-time fluorescence quantification PCR combined with Sanger sequencing was established ,which had high sensitivi-ty(101copies/μL)andgoodrepeatability(intra-assayCVandinter-assayCVofthereal-timefluorescencequantificationPCRofex-on 19 and 21 were 1 .42% /3 .52% and 0 .97% /2 .44% ,respectively ) .20 clinical samples were simultaneously detected by this method and the traditional Sanger sequencing ,the results were completely consistent .Conclusion The EGFR gene mutations detec-tion method of the real-time fluorescence quantification PCR combined with Sanger sequencing is successfully established ,which can be used in the clinical sample detection .

A new method for the determination of peptide antibiotics in sediment from aquaculture environment by high performance liquid chromatography-tandem mass spectrometry was developed. The target analytes in sediments were ultrasonically extracted twice with citrate buffer solution and methol mixture (3∶ 4, V/ V), followed by complexation with 0. 5 g of Na2 EDTA, purification with 5 mL of methyl isobutyl ketone, and clean-up with HLB-SPE column. The analytes were separated on a MGII C18 column by gradient elution with 0. 1% formaic acid-0. 1% formaic acid acetonitrile as mobile phase, detected in multiple reaction monitoring (MRM) with electrospray ionization (ESI) under positive ion mode, and quantified by external standard method. The calibration curves were linear (R2 >0. 999) over a concentration range of 10 -10000μg / L for colistin and bacitracin and 4-4000 μg / L for virginiamycin M1 . The limits of detection (S / N = 3) were 5 μg / kg for colistin and bacitracin and 2 μg / kg for virginiamycin M1 . The limits of quantification (S / N=10) was 10 μg / kg for colistin and bacitracin and 4 μg / kg for virginiamycin M1 . At three spiked levels, the recoveries ranged from 79. 7% to 91. 6% (RSD=1. 9% -10. 8% ), showing high sensitivity, good reproducibility and wide applicability.

Clinical practice is an important part for medical students to become qualified doctors.The humanistic skill and medical operation skill training,test and evaluation before clinical practice are necessary.Results showed that the standard and routine training before clinical practice can help interns adapt to their practice life more easily.

Aim To investigate the effect of oral itraconazole on pharmacokinetics of viaminate and under-stand the metabolism pathway of viaminate in healthy humans,as well as guide the reasonable medication for viaminate and itraconazole coadministration.Methods The study was conducted in a double-blind randomized and crossover manner with two phases of treatment with placebo-viaminate or itraconazole-viaminate.Eight healthy male subjects received an oral dose of 200 mg itraconazole(QD) or matched placebo for five consecutive days,and took an oral 100 mg dose of viaminate on day 4 after each treatment phase.The liquid chromatography-mass spectrometry(LC-MS) was established to measure the blood drug concentration of viaminate.Results Our results showed that the mean peak concentration of viaminate was increased with itraconazole [(21.21?11.65) ?g?L~(-1) vs(27.12?13.83) ?g?L~(-1),P0.05].Additionally,the mean elimination half life was prolonged [(2.71?0.38) h vs(4.43?0.93) h,P

Objective To observe the pituitary tumor transforming gene (PTTG), estradiol-stimulated protein 1 (ESP1) expressions in S, G 2/M phases in A549, SPC-A-1 cell lines. Methods The mammalian cells for cell cycle was synchronizated by thymidine, the total RNA in the S, G 2/M phases was collected, and the total RNA was observed by Real-time PCR. Results PTTG expression in A549 and SPC-A-1 cell lines was higher in S phase than that in G 2/M phase while no PTTG expression in MRC-5 cells in S, G 2/M phases. ESP1 expression in A549, SPC-A-1, MRC-5 cell lines was lower in S phase than that in G 2/M phase. Conclusion That no PTTG in S and G 2/M phases in the normal cell line and PTTG expression in the lung cancer cell line higher in S phase than that in G 2/M phase suggested that the aneuploidy may be caused by the overexpression of PTTG. ESP1 expression was lower in S phase than G 2/M phase in all of the three cell lines, suggesting that sister chromatids separation depend on the active ESP1.

Objective To investigate the relationship between day and night regularity of asymptomatic ventricular presystole (VPS) and the activity of autonomic nervous system and its clinical significance Methods Thirty children with the absence of underlying organic heart disease were examined with 24 hour dynamic electrocardiogram (DCG), the function of autonomic nervous system and ventricular late potentials (VLP) Results The rate (43%, 13/30) was the day type VPS, 12 out of the 13 resulted from increased activity of sympathetic nerve The rate (30%, 10/30) was the night type VPS, all of them resulted from the activity of the vagus nerve enhanced There was a positive correlation between VPS frequency of day type and heart rate ( r =0 67, P

Objective To explore the effects of the therapeutic effects of interventional heated chemotherapy( IHC) for moderate and advanced huge primary hepatocellular carcinoma and compare with transcatheter arterial chemoembolization( TACE) . Methods The 68 pa-tients with moderate and advanced huge primary hepatocellular carcinoma accepted the treatment of IHC were considered as the observation group,and 32 patients managed TACE were regarded as the control group. The therapeutic effects,living quality,secondary surgery rates,The reduced level of serum AFP,survival rate,and toxic side effects of 2 groups were contrasted. Results The effective rate of observation group was obviously higher than that of control group (79. 4% vs 56. 3%,P <0. 05),and secondary surgery rates was increased(70. 6% vs 43. 8%,P<0. 01). The ratio of increased Karnofsky score was more than 20 in observation group,which was obviously higher than that of control group(35. 3% vs 12. 5%,P<0. 01). T The reduced level of serum AFP was more than 50% in observation group,which was higher than that of control group (52. 9% vs 25. 0%,P<0. 01). The survival rate of 12,18,24 months after treatment in control group was in-creased than that in control group individually (P<0. 01). Conclusion IHC is effective for huge primary hepatocellular carcinoma and in-creases the secondary surgery rates,which is one of the safe and effective methods for huge primary hepatocellular carcinoma.

Objective:To design ultrasonic atomization device making use of disposable water cup, to improve ventilation therapy in clinical application.Methods: The design take advantage of working principal of ultrasonic atomization, do a simple interface compatible work to the disposable water cup, the water cup can realize the ability of medicine atomization treatment, collecting the water when not do atomization; the water cup access into the ventilator pipe, to test the influence between the atomization and ventilator function.Results: The disposable water cup realized ultrasonic atomization successfully, and shows no influence between the atomization and ventilator self-function.Conclusion: The design has made some simple improvement of the disposable water cup on the structure, proposed a new way of ventilator atomization practically and economically.

Objective To elucidate the correlation between Chinese patients with Crohn's disease (CD) and Mycobacterium avium subspecies paratuberculosis (MAP) infection.Methods The peripheral blood of 73 patients with CD (ileocolon type) and 40 healthy individuals was obtained.The monocyte layer was isolated for MAP culture.The growth of bacteria colony in the culture tube was observed every two weeks and totally for 16 weeks.By the end of culture,specific DNA of MAP (IS900) in the culture tube was detected by nested polymerase chain reaction.ANOVA was performed for measurement data analysis and chi square test was used for count data.Results After cultured for 16 weeks,no bacteria colony of MAP had been observed in 113 mycobacteral growth indicator tube (MGIT).No IS900 had been detected in the peripheral blood of all patients with CD and all healthy controls.Conclusion MAP is not correlated with the pathogenesis of CD (ileocolon type) in Chinese patients.