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 .

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 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 .

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.

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 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 observe the hindpaw withdrawal threshold (HWT), and the ultrastructure and expression of glia cell line-de-rived neurotrophic factor (GDNF) in sciatic nerve (SN) in chronic constriction injury (CCI) rats after pulsed radiofrequency (PRF). Meth-ods 30 Sprague-Dawley rats were divided into sham modeling-sham treating (SS) group, CCI-Sham treating (CS) group and CCI-PRF (CP) group. The right SNs of the rats in the CS and CP groups were ligated, and it was separated without ligation in the SS group. The CP group accepted PRF at the ligation 14 days after modeling, while the electrodes were placed without electricity in the SS and CS groups. Their HWT was measured before and 1, 7, 14 days after modeling, and 1, 7, 14 days after treatment. The right SN of ligation was observed under electron microscope 14 days after treatment, meanwhile, the GDNF expression was determined with enzyme-linked immunosorbent assay (ELISA). Results HWT was significantly shorter in the CS and CP groups than in the SS group after modeling, and it increased in the CP group 14 days after treatment compared with that of the CS group (P<0.01). The degeneration of SN significantly improved in the CP group compared with the CS group, while the expression of GDNF increased compared with that in the CS and SS groups (P<0.01). Conclusion PRF could relieve the CCI-induced neuropathic pain by upregulating the GDNF expression in the SN to prevent the SN from injury.

To improve the quality of teaching is the core task of the development of higher education,and effective teaching is the fundamental guarantee to promote students to learn effectively,and to improve the teaching quality and talents cultivation quality,which has very important significance to exploring the effective teaching idea and implementation strategy.In the teaching of medical immunology for example,the author combined the characteristics of the course,and had a preliminary discussion on the feature of effective teaching,to explore the effective teaching strategies of thought guiding before class,creating context in class,autonomous learning at recess,teaching reflection after class,to provide reference for college teachers to improve teaching ability and teaching effectiveness.