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.

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 study the expression of HDAC2 in lung of smoke cessation and smoking cessation rats,Methods Forty SD rats were divided into five groups randomly:normal control group (group A),1-month smoking group (group B,),2-month smoking group (group C),1-month smokingcessation group(group D),2-month smoking-cessation group(group E).Group B rats were exposured to cigarettes for 1 month,C,D,E were exposured to cigarettes for 2 months.At 1 month,group B were sacrificed,at 2 month,group C were sacrificed,group D was smoking cessation for 1 month and group E for 2 months.Pathomorphological changes of the small airway were analyzed,and then study the HDAC2 in rats'lung tissue.Results Compared with group A,the levels of HDAC2 in group B,group C,group D,group E were decreased (P ＜ 0.01),but smoking groups levels were lower than smoking-cessation groups.In smoking groups,group C was lower than group B.In smoking-cessation groups,group D was lower than group E.Conclusion It shows that the levels of HDAC2 in rats' lung tissue decrease after smoking exposure.It can recrease after quitting,but still cant back to normal.

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.

0.05),but those in severe hyperbilirubinemia group significantly higher(Pa

Objective To explore the early diagnostic value of the expression level of plasma N-terminal pro-B-type natriuretic peptide(NT-proBNP)for heart failure(HF)in neonates.Methods Thirty-five neonates who were clinic diagnoses as HF newborns and 20 cases of non-HF newborns(control group)were selected,on the 2nd,the 7th day after birth,plasma NT-proBNP and CK-MB levels were measured with electrochemiluminescence method and mass method.All data were analyzed by SPSS 16.0 software.Results Compared with the control group,CK-MB and NT-proBNP were higher in the HF group before treatment(P0.05).Compared with before treatment,CK-MB and NT-proBNP were significantly lower in the HF group(P0.05),NT-proBNP level was lower in the control group(P

Objective To understand the clinical characteristics of primary small vessel vasculitis with involvement of lungs in children and analyze the causes of misdiagnosis.Methods Clinical data from 16 patients with primary small vessel vasculitis were collected and their pathological,antineutrophil cytoplasmic antibody(ANCA),pulmonary lesion manifestations,image and misdiagnosis were further analyzed.Results Among 16 cases,9 cases were microscopic polyarateritis(MPA) and 4 cases were Wegeners granulomatosis(WG) and 3 cases were Churg-Strause syndrome(CSS).The ages of onset were from 3 to 14 years old with average age of 7.5 years old.Seventy-five percent(12/16) were ANCA positive.The patients with lung involvement had cough,chest pain,dyspnea,hemoptasis and asthma.Findings on lung X-ray films were variable.Some showed multiple patch shadows,others showed single or multiple masses or nodular shadows with or without cavity formation.After the treatments with corticosteroid and cyclophosphamide,10 cases turned better;including 8 cases with normal on lung X-ray films.Three cases given up;3 cases died.Conclusion It is very difficult to make diagnosis of primary small vessel vasculitis with involvement of lungs and should be paid more attentions to this issue.