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.

Antidepressive Agents ; therapeutic use ; Depression ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Electroacupuncture ; Humans ; Integrative Medicine ; Phytotherapy

Objective To explore the relationship between the genetic relationship and the cross compatibility of the wild rootstocks which belong to malus mill plants.Methods With the antecedent study in the cross compatibility of the rootstocks of malus mill plants, this paper focuses on the nrDNA ITS sequence analysis in the samples of the wild rootstocks which belong to malus mill plants from different 24 regions of China.Results It was successfully that having measured the sequence of 24 samples and having made the genetic distance matrix and genetic tree diagram.Conclusion There is a certain relationship between the genetic relationship and the cross compatibility of the wild rootstocks which belong to malus mill plants.

Objective To evaluate the effect of evidence based nursing on the prevention of the complications of PICC .Meth‐ods The randomised controlled trials (RCT) on the evidence based nursing in reducing or preventing the complications of patients with PICC were collected using the databases of PubMed ,CNKI ,WanFang ,CBM and VIP;the data were analyzed by RevMan5 .0 software .Results 17 studies involving 3 092 patients were included in total .The results of Meta analyses showed that compared with the control group ,the complication (Phlebitis ,puncture point infection ,catheter prolapse or ectopic and catheter obstruction) rates of evidence based nursing group with PICC were lower (P<0 .05) than that of the control group .Conclusion Evidence based nursing can effectively prevent the complication of PICC ,improve the quality of nursing and worth of clinical application .

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.

Objective To explore the effect of Renzhu Jianwei Granules(Granule of Semen Coicis and Rhizoma Atractylodis Macro- cephalae for strengthening the stomach)on the patients with precancerous lesions of gastric cancer(PLGC)and its mechanism. Methods The 58 patients of chronic atrophic gastritis(CAG)complicated with intestinal metaplasia(IM)and/or dysplasia(Dys) were randomized into treatment group and control group with 29 in each.The treatment group was administered Renzhu Jianwei Gran- ules while the control group was given Weifuchun Tablets.The numbers of positive cells of carcinoembryonic antigen(CEA)and cy- clooxygenase 2(COX-2)of gastric mueosa before and after treatment were observed.Results The scores of positive cells of CEA and COX-2 of gastric mucosa of both groups were greatly reduced after treatment(P0.05).Conclusion Renzhu Jianwei Granules can reduce CEA and COX-2 of PLGC patients,possibly being one of the mechanisms in treating PLGC.

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.