Objective To observe the missed diagnosis of invasive carcinoma under the microscope (ICUM) in high grade squamous intraepithelial neoplasia (HSIL), and analyze associated factors influencing missed ICUM. Methods A retrospective study was performed on patients diagnosed with HSIL by colposcopy-guided biopsy and treated with loop electrosurgical excision procedure(LEEP)at the First Affiliated Hospital of Nanjing Medical University, from December 2014 to December 2016. They were non-pregnant, ≤50 years old and the cervical volume without obvious enlargement and exogenous surface without and ulcerative lesions. A total of 283 cases with early cervical cytology results, never received cervical traumatic treatment or cervical biopsy in another hospital before, and their colposcopic images were clear enough to reevaluate. The ultimate pathological diagnosis was based on the higher-level pathological diagnosis between the results of cervical biopsy and LEEP to evaluate ICUM missed in HSIL and the risk factors. Results (1) Among the 283 cases with HSIL diagnosed by colposcopy-directed biopsy,44 cases (15.5％,44/283) were missed diagnosis of ICUM, which consisted of 29 cases Ⅰa1, 4 cases Ⅰa2 and 11 cases Ⅰb1 in the ultimate pathology.(2)Analysis of associated factors for missed ICUM:univariate analysis showed that,as the age increased, the risk of missed ICUM also increased(the rates of missed diagnosis for<30, 30-39, 40-50 years were 7.7％, 11.5％, 22.0％, respectively;χ2=6.254, P=0.012 by trend test). The more the number of high-grade features, the higher risks(the rates of missed diagnosis for 1, 2, 3, 4 high-grade features were 10.2％, 17.6％, 23.8％, 30.8％, respectively;χ2=7.686,P=0.006 by trend test). The locations of HSIL were only endocervical, only ectocervical and mixed, the risk increased by this sequence(2.8％, 5.1％, 28.7％; χ2=26.193,P<0.01 by trend test). The rate of missed diagnosis for not completely visible squamocolumnar junction(SCJ)was higher than that of the completely visible one(22.3％ vs 2.1％;χ2=19.680, P<0.01). The rate of missed diagnosis was higher for existing atypical vessels than those without(60.7％ vs 10.6％;χ2=48.279, P<0.01). The rate of missed diagnosis for visible lesion size≥40 mm2 was higher than that of<40 mm2(27.3％vs 4.2％;χ2=28.921, P<0.01). The rate of missed diagnosis for the proportion of visible lesion size in ectocervical size ≥0.75 was higher than that of <0.75 (83.3％ vs 14.1％;P<0.01). The rate of missed diagnosis for the maximum linear length of visible lesion≥10 mm was higher than that of<10 mm(46.9％vs 9.0％;χ2=44.473, P<0.01). But the different severity of cervical cytology before colposcopy was not associated with missed ICUM(P>0.05). Multivariable analysis found that visibility of SCJ, atypical vessels, visible lesion size and maximum linear length of visible lesion were associated with missed diagnosis of ICUM(all P<0.05). Conclusions The diagnostic value of HSIL by colposcopy is limited. Meanwhile, for the patients who are ≤50 years old with HSIL diagnosed by cervical biopsy, invisibility of SCJ, atypical vessels, visible lesion size and maximum linear length of visible lesion evaluated by colposcopy are the independent risk factors of missed ICUM. Thereby, it is necessary to take active intervention for HSIL with these risk factors.

Attention deficit/hyperactivity disorder (ADHD) is one of the most common neuro developmental disorders in childhood.The main symptoms of ADHD are inattention, hyperactivity, impulsivity, etc.ADHD causes lots of adverse effects on patients, families and the society.Currently, medication is a common treatment for ADHD.However, due to a series of adverse effects caused by drug treatment, and some concomitant problems, the effect of drug treatment alone is not ideal.Therefore, non-drug treatment of ADHD is becoming popular.This study aims to review the common non-drug treatment methods and efficacy of ADHD at home and abroad.

Objective:To explore the effects of mutual-aid training mode in nursing interns.Methods:From June 2017 to June 2019, this study selected 70 nursing interns as subjects in Department of Acupuncture and Rehabilitation of Jiangsu Provincial Hospital of Chinese Medicine by convenience sampling. Nursing interns from June 2017 to June 2018 were in control group ( n=35) , and those from July 2018 to June 2019 were in observation group ( n=35) . Nursing interns of control group received the traditional training mode, while observation group carried out the mutual-aid training mode. The study compared the theoretical achievement, operational performance, clinical thinking ability and teaching effect. Results:After training, the theoretical achievement and operational performance of nursing students in observation group were higher than those in control group with statistical differences ( P<0.05) ; the total score and dimension scores of clinical thinking ability of nursing students in observation group were higher than those in control group with statistical differences ( P<0.05) ; the teaching effect evaluation of nursing students in observation group was better than that in control group with a statistical difference ( P<0.05) . Conclusions:The mutual-aid training mode can improve nursing students' clinical nursing skills and nursing quality which is worthy of clinical popularization.

Objective: To establish a method for detection of chikungunya virus(CHIKV) antigen. Methods: CHIKV virus like particle(VLP), that contains all structural proteins, was prepared by baculovirus expression system. Mice and rabbits were immunized with the VLP to develop antibodies against CHIKV. A double antibody sandwich ELISA was established for detection of CHIKV antigens. The concentrations of the antibodies used and the reaction conditions were optimized. The detection limit and repeatability of the ELISA was evaluated. Results: The sensitivity and specificity was estimated by 10 mimicking CHIKV sera, 90 health person sera, 40 other virus infected sera. It was show that the specificity of DAS-ELISA was 100%, the detection limit was 10 TCID₅₀, the coefficients of variation (CV) within plate was <5%, the CV of different plates was <10%. Conclusions The double antibody sandwich ELISA established in this study can be used to detect the CHIKV antigen in acute phase serum of patient and provide a method for detection of CHIKV.

In order to understand the prevalence and genetic variation of H9N2 subtype avian influ-enza virus in Shandong,a total 492 tracheal and lung tissue samples collected from chicken farms with respiratory symptoms in partial areas in Shandong were detected by H9 subtype AIV real-time RT-PCR,and the positive samples were inoculated with chicken embryos for two generations.Whole genome sequences of the positive strains by applying Illumina Miaseq platform,and genetic evolution and mutation at positions associating with viral pathogenicity and transmissibility were analyzed.The results showed that there were 72 samples were positive for H9 subtype AIV among the 492 samples,with a positive rate of 14.63％.Thirty-four strains of H9 subtype AIV were ob-tained from the positive samples after passing through chicken embryo,meanwhile,the 34 isolates were all H9N2 subtype AIV by whole genome sequencing analysis.By analyzing the evolutionary tree of HA and NA genes,HA and NA genes of the 34 H9N2 AIV strains belonged to Y280-like branch and F/98-like branch,respectively.Meanwhile,based on above branches,there were obvious time node subbranch,which one was"isolates before 2013",another one was"isolates after 2013".The HA cleavage sites of thirty-four H9N2 strains were all 325PSRSSR↓GLF333,which met the se-quence characteristics of the lowly pathogenic avian influenza virus,and the HA receptor binding site 226 amino acid was leucine,which had the characteristics of blinding to a-2,6 mammalian sialic acid receptors.Among the internal amino acid sites that are key to mammalian adaptation,all strains had an I368V mutation in the PB1 gene that enhanced viral transmissibility in mammals and the PB2 genes of some strains were mutated to enhance the mammalian adaptation of I292 V and A588 V.The above results illustrated that the H9N2 subtype AIV gene segments in Shandong have different degrees of recombination and gene variation,so it is necessary to strengthen the monito-ring of virus variation.

Objective:To investigate the molecular testing of liquid-based cytology (LBC) specimens from advanced non-small cell lung cancer (NSCLC) patients and the reliability of guiding targeted therapy.Methods:The LBC specimens and clinical data of 412 advanced NSCLC patients from March 2015 to April 2017 in the Cancer Hospital, Chinese Academy of Medical Sciences were collected, of which 32 patients had postoperative or biopsy specimens. The real-time quantitative polymerase chain reaction was used to detect mutations of EGFR, KRAS and BRAF, and analyze the correlation between gene mutations and clinicopathological characteristics. The results of genetic testing of LBC specimens and histology specimens were examined for concordance. Clinical efficacy was evaluated in 142 patients treated with EGFR-tyrosine kinase inhibitor (TKI) drugs, and survival analysis was performed using the Kaplan-Meier method.Results:Of the 412 LBC specimens, 216 (52.4%) had EGFR mutations, 36 (8.7%) had KRAS gene mutations, and 3 (0.7%) had BRAF gene mutations. EGFR mutation was associated with gender, pathology type, and specimen source, with a higher EGFR mutation rate in female patients (63.0%) than in male patients (40.8%, P<0.001) and a higher EGFR mutation rate in adenocarcinoma (54.3%) than in non-adenocarcinoma (0.0%, P<0.001). KRAS mutation was related to gender, with a higher EGFR mutation rate in male patients (12.2%) than in female patients (5.6%, P=0.016). The three cases with multiple co-mutations were all stage Ⅳ male adenocarcinoma patients. Thirty-two patients with both LBC specimens and histology specimens had concordant genetic results between LBC specimens and histology specimens in 30 patients ( Kappa=0.91). Twelve patients with both histology and LBC specimens from metastases had identical genetic results ( Kappa=1.00). Nineteen patients with histology specimens from primary foci in lungs and LBC specimens from metastases had concordant genetic results between two specimens in 18 patients ( Kappa=0.92). The disease control rate (DCR) for EGFR mutation-positive patients treated with EGFR-TKI was 89.0% (89/100) and the progression-free survival time (PFS) was 13.8 months, both higher than those of EGFR mutation-negative patients [DCR of 30.8% (4/13) and median PFS of 1.4 months, P<0.01]. Conclusions:The results of molecular testing of LBC specimens and histological specimens are highly consistent, which demonstrates LBC specimens can be a crucial source of gene testing for advanced NSCLC. Molecular typing of advanced NSCLC based on the results of genetic testing of LBC specimens and guiding EGFR-TKI drug-targeted therapy can achieve high DCR and PFS, which has important clinical value.

Objective:To investigate the molecular testing of liquid-based cytology (LBC) specimens from advanced non-small cell lung cancer (NSCLC) patients and the reliability of guiding targeted therapy.Methods:The LBC specimens and clinical data of 412 advanced NSCLC patients from March 2015 to April 2017 in the Cancer Hospital, Chinese Academy of Medical Sciences were collected, of which 32 patients had postoperative or biopsy specimens. The real-time quantitative polymerase chain reaction was used to detect mutations of EGFR, KRAS and BRAF, and analyze the correlation between gene mutations and clinicopathological characteristics. The results of genetic testing of LBC specimens and histology specimens were examined for concordance. Clinical efficacy was evaluated in 142 patients treated with EGFR-tyrosine kinase inhibitor (TKI) drugs, and survival analysis was performed using the Kaplan-Meier method.Results:Of the 412 LBC specimens, 216 (52.4%) had EGFR mutations, 36 (8.7%) had KRAS gene mutations, and 3 (0.7%) had BRAF gene mutations. EGFR mutation was associated with gender, pathology type, and specimen source, with a higher EGFR mutation rate in female patients (63.0%) than in male patients (40.8%, P<0.001) and a higher EGFR mutation rate in adenocarcinoma (54.3%) than in non-adenocarcinoma (0.0%, P<0.001). KRAS mutation was related to gender, with a higher EGFR mutation rate in male patients (12.2%) than in female patients (5.6%, P=0.016). The three cases with multiple co-mutations were all stage Ⅳ male adenocarcinoma patients. Thirty-two patients with both LBC specimens and histology specimens had concordant genetic results between LBC specimens and histology specimens in 30 patients ( Kappa=0.91). Twelve patients with both histology and LBC specimens from metastases had identical genetic results ( Kappa=1.00). Nineteen patients with histology specimens from primary foci in lungs and LBC specimens from metastases had concordant genetic results between two specimens in 18 patients ( Kappa=0.92). The disease control rate (DCR) for EGFR mutation-positive patients treated with EGFR-TKI was 89.0% (89/100) and the progression-free survival time (PFS) was 13.8 months, both higher than those of EGFR mutation-negative patients [DCR of 30.8% (4/13) and median PFS of 1.4 months, P<0.01]. Conclusions:The results of molecular testing of LBC specimens and histological specimens are highly consistent, which demonstrates LBC specimens can be a crucial source of gene testing for advanced NSCLC. Molecular typing of advanced NSCLC based on the results of genetic testing of LBC specimens and guiding EGFR-TKI drug-targeted therapy can achieve high DCR and PFS, which has important clinical value.