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.

OBJECTIVETo explore the role of traditional Chinese medicine (TCM) syndrome scores in effectiveness evaluation of clinical studies.

METHODSRandomized controlled trials (RCTs) of TCM published in five journals in 2013 were retrieved, including Journal of Traditional Chinese Medicine (JTCM), Chinese Journal of Integrated Traditional and Western Medicine (CJITWM), Chinese Journal of Integrative Medicine (CJIM), Evidence-Based Complementary and Alternative Medicine (ECAM), and American Journal of Chinese Medicine (AJCM). The details of TCM syndrome scores and other relevant factors reported in articles were extracted and analyzed. Descriptive statistics and Chi-square test were used to describe general features of inclusive studies, ratios of reports on CM syndrome scores in each journal, formulated evidence, adopted evaluation rules, important degrees. The difference in the application rate of CM syndrome scores were compared in various diseases, diseases with or without CM syndrome typing, places where clinical studies were implemented, and different journals.

RESULTSA total of 240 RCTs were included, involving 178 published in Chinese version and 62 in English version. CM syndrome scores were used for effectiveness evaluation in 27.1% (65/240) of RCTs, of which, the highest application percentage was 35.3% (18/51) in RCTs published in CJITWM, and the lowest was 0 (0/7) in RCTs published in AJCM. There were 17 methods for grading TCM syndrome scores, of which Guideline for Clinical Research of New Chinese Herbal Medicine was most commonly used. Detailed grading standards for CM syndrome scores were reported in 46 RCTs, and CM syndrome scores were taken as primary or secondary outcomes in 6 RCTs. When describing the percentages of RCTs adopting TCM syndrome score by diseases classification, the lowest was 9.5% (2/21) in mental and behavioral disorders, and the highest was 39.1% (9/23) in endocrine, nutritional, and metabolic diseases. RCTs with TCM syndrome differentiation had a higher percentage of adopting TCM syndrome score than those without TCM syndrome differentiation (P < 0.01). Statistical differences were also found among RCTs published in Chinese core journals (32.0%), domestic science citation index (SCI) journals (28.6%), and overseas SCI journals (4.9%) (χ2 = 12.4593, P < 0.01). For RCTs conducted by Chinese scholars, the percentage of using TCM syndrome score was 32.0% in three journals from China, while 7.1% in two foreign language journals, showing significant difference (χ2 =7.3615, P < 0.01); none of RCTs conducted by foreign scholars used TCM syndrome scores for effectiveness assessment.

CONCLUSIONSThere was a lack of agreeable and universal standards for TCM syndrome scores using in effectiveness evaluation. Therefore, it was not commonly used in domestic CM clinical studies. It was not so favorably agreed in overseas academic circles.

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

Liver transplantation is the only definitive treatment modality of end stage liver diseases. Demyelinative disease is rarely seen in patients after liver transplantation, but a higher incidence has been noticed recently. The disease is liable to be misdiagnosed as immunosuppressant-induced psychiatric disorders at early stage. Central pontine myelinolysis is more disastrous and it will greatly influence the short-term survival and long-term life quality of the patients after liver transplantation. When it is manifested as peripheral nervous system disorder, the disease usually has a subtle onset, making it difficult for diagnosis and treatment. The causes for various demyelinative diseases should be understood to prevent it in patients after liver transplantation. The specific mechanism for demyelinative disease after liver transplantation remains unclear. We introduce the possible causes of common demyelinative diseases, hoping to provide reference for prevention and treatment of such conditions after liver transplantation.

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.

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

The models of Cardiacglycoside-induced cardiotoxicity in guinea pigs were made by perfusing isolated heart with Langendorff appa- ratus. The free radicals generation in myocardium following the incidence of ventricular premative beat, ventricular tachycardia and ventricular fibrillation were measured respectively by the directed measurement with Electron Spin Resonance ( ESR ) techniques. The results showed the more severe toxicosis, the more much free radicals generate. Free radicals generated significantly in ventricular fibrillation and were scavenged by Superoxide dismutase ( SOD ) and sodium selenate.

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.