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 .

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

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.

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.

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 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