Background Research comfirmed that second mitochondrial activator of caspase (Smac) is a promoting tumor cell apoptosis protein.Our previous study showed that the expression level of Smac in LECs is obviously higher in cataract than that in normal eyes.We assumed that silencing Smac gene in LECs can inhibit the apoptosis of LECs.The way to transfect Smac siRNA into LECs is a key step.Objective This study was to construct siRNA lentiviral vector of Smac and identify its silencing efficiency in human lens epithelial B3 cell line (HLE-B3) and establish low-expressed Smac HLE-B3 line.Methods Based on the genebank and our previous study,siRNA sequence of Smac was designed and composed.The synthetic double-stranded DNA was linked to the lentiviral vector GVll8 by T4 DNA ligase and then transformed DH5α competent cells.The plasmids were transformed into the DH5α competent cells.Recombinant colonies were screened by PCR and sequenced.Recombinant plasmids and two other auxiliary plasmids were used to infect 293T cells.Cell culture supernatant was collected for the measurement of viral titer.Recombinant lentiviral vector was used to infect HLE-B3 cells to calculate the viral multiplicity of infection (MOI) under the fluorescence microscope.Transfection efficiency was examined by calculating the GFP-positive cells.HLE-B3 cells were divided into negative control group,siRNA plasmid tranfected group and GV118-Smac-siRNA1 tranfected group.The relative expression levels of Smac mRNA in the cells were detected and compared among the three groups by real-time fluorescent quantitative PCR.Results GV118-Smac-siRNA was successfully constructed with the positive colonies 340 bp and blank vector colonies 299 bp,and viral titer was 3.0× 108 TU/ml.At a MOI of 100,the infecting efficiency of the vector on HLE-B3 cells was about 82％ and the cytotoxicity was low.The relative expression levels of Smac mRNA were (101.290±8.349)％,(92.330±6.320)％ and (32.540±4.221)％ in the negative control group,siRNA plasmid tranfected group and GV118-Smac-siRNA1 tranfected group,respectively,showing a significant difference among the three groups(F =32.871,P＜0.01),and the relative expression level of Smac mRNA was significantly lower in the GV118-Smac-siRNA1 tranfected group than that in the negative control group (P =0.000).However,no significant difference was found in the Smac mRNA expression between the blank plasmid group and the negative control group (P=0.535).Conclusions GV118-Smac-siRNA lentiviral vector is successfully constructed.Smac-siRNA can effectively inhibit the expression of Smac mRNA in human LECs.

Objective To analyze relevant clinical and dosimetric factors associated with radiation pneumonitis in patients with stage Ⅲ non-small cell lung cancer after they received radical radiotherapy.Methods A total of 126 patients with stage Ⅲ non-small cell lung cancer who received precision radiotherapy in Peking University Cancer Hospital were analyzed from January 2013 to December 2014.Data were collected including various clinical factors (including sex,age,histological type,tumor location,history of diabetes,history of hypertension,history of smoking,the season patients received treatment,ECOG performance status before treatment,chemotherapy before radiotherapy,concurrent chemotherapy and the classification of radiation pneumonitis),as well as related dosimetric parameters [including GTV,lung volume (LV),bilateral V5,V10,V20,V30 and MLD].SPSS 19.0 software was used to analyze the relation between correlation factors and radiation pneumonitis (RP≥2).Results Among the patients,31 cases (24.6％) had occurrance of radiation pneumonitis ≥ 2.Univariate analysis showed that age,ECOG performance status before treatment,concurrent chemotherapy and GTV/LV ratio were significantly correlated with RP ≥ 2 (R =0.157-0.222,P ＜ 0.05).Further multivariate Logistic regression showed that age,concurrent chemotherapy and GTV/LV ratio were significantly correlated (Wald =4.754,6.422,14.79,P ＜ 0.05).Conclusions In patients with stage Ⅲ non-small cell lung cancer after receiving thoracic radical radiotherapy,increasing age and GTV/LV ratio≥3.2％ are risk factors of RP≥2.The concurrent chemotherapy with low-dose paclitaxel might also increase the risk of RP≥2.

Objective:To understand the patients'satisfaction on catastrophic medical insurance of New Rural Cooperative Medical Scheme ( NRCMS ) , and provide references for effective management and policy improvement. Methods:According to the principles of representative sampling, a questionnaire survey conducted. 484 patients got compensation for catastrophic insurance in 2013 were surveyed, and information of medical costs was analyzed. Re-sults:The average medical cost accounts for 84. 57% of total household expenditure. Mandatory medical expenditure is still high and economic burden of diseases is heavy. This policy has increased the compensation rate, and the actu-al compensation rate increases 4. 79% in three districts. The average score of patients' attitude toward catastrophic medical insurance is (3.95±1.05) points, 88. 43% of patients continue to participate in medical insurance. Con-clusions and suggestions:Patients' satisfaction on catastrophic medical insurance of NRCMS is high. It is suggested that the existing financing modes should maintain stability in the short term, and the compensatory rate should be im-proved reasonably.

Objective To compare the dosimetric difference between RapidArc and fixed gantry angle dynamic IMRT (dIMRT) for central-type lung cancer radiotherapy. Methods Therapy for 10 patients previously treated with dIMRT was replanned with RapidArc. Dose prescription was 66 Gy/33 fraction. Comparative endpoints were planning target volume (PTV) dose, doses to surrounding structures,number of monitor units, and treatment delivery time. Results There was no significant dosimetric difference between RapidArc and dIMRT. Compared with dIMRT, RapidArc slightly elevated target volume dose, lung V5, V10. The average values of lung V20, V30 and heart V30 were larger in dIMRT than those in RapidArc. The number of monitor units was reduced by 32% and the treatment time by 66% in RapidArc.Conclusions Both RapidArc and dIMRT plans could meet the clinical therapy needs. RapidArc could achieve similar target coverage and sparing of organs at risk, with fewer monitor units and shorter delivery time than dIMRT.

Objective To explore the directively meaning of Hans Selye ’ s discovery of biological stress to ethical issues facing donor families after cardiac death , and to alleviate the anxiety of donor families and get more organs for transplantation .Methods The data from 54 patients with potential donors in the First Hospital of Jilin University were collected , and the sources of psychological pressure in the donor families were analyzed , and the effect of Hans Selye ’ s discovery of biological stress on relieving the pressure and strengthening the coping with the ethical issues facing donor families were explored .Results Thirty-five patients with potential donors in 54 cases agreed with donors , and 22 patients with potential donors were successful donation at last . Conclusions Hans Selye’s discovery of biological stress can be used in the management of donor families after cardiac death in order to provide the security for striving for more transplantable organs .

Objective To investigate the consensus and controversies on the delineation of radiotherapy target volume for patients with locally advanced non-small cell lung cancer (LA-NSCLC).Methods Questionnaires including 15 questions on the delineation of radiotherapy target volume of NSCLC were sent to 12 radiation departments in China in November 2015.A patient with LA-NSCLC was selected by Fudan University Shanghai Cancer Center, and simulation CT images and medical history data were sent to the 12 radiation departments.Twelve radiation oncologists from the 12 radiation departments showed and explained the delineation of radiotherapy target volume of their own, and the patient was discussed by all experts in the sixth multidisciplinary summit forum of precise radiotherapy and chemotherapy for tumor and lung cancer.Results All receivers of the questionnaire answered the questions.The standard lung window width/level for the delineation of lung cancer was 800-1600/-600 to-750 HU, and the mediastinum window was 350-400/20-40 HU.Respiratory movement was measured by stimulator, 4D-CT, and stimulator+4D-CT with 2-5 mm expansion based on experience.The primary clinical target volume (CTV) was defined as gross target volume (GTV) plus 5-6 mm for squamous carcinoma/5-8 mm for adenocarcinoma.The metastatic lesion of mediastinal lymph nodes was delineated as 5 mm plus primary lesion in 6 departments and as primary lesion in another 6 departments.Of the 12 departments, 10 applied 5 mm of set-up error, 1 applied 3 mm, and 1 applied 4-6 mm.For V20 of the lungs, 10 departments defined it as<30%, 1 as<35%, and 1 as 28%.Nine departments defined the radiation dose of concurrent chemoradiotherapy (CCRT) for LA-NSCLC as 60 Gy in 30 fractions, 62.7 Gy in 33 fractions in 1 department, 50-60 Gy in 25-30 fractions in 1 department, and 60-70 Gy in 25-30 fractions in 1 department.For the delineation of target volume for the LA-NSCLC patient treated with CCRT, the primary planning target volume (PTV) was defined as GTV plus organ movement (IGTV) and set-up error (GTV→IGTV→PTV) in 3 departments, as CTV plus organ movement (ITV) and set-up error (GTV→CTV→ITV→PTV) in 8 departments, and as CTV plus set-up error/IGTV plus 5-6 mm for squamous carcinoma/5-8 mm for adenocarcinoma (CTV) and set-up error (GTV→CTV→PTV/GTV→IGTV→CTV→PTV) in 1 department.For the delineation of PTV in the mediastinal lymph node, GTV→IGTV→PTV was performed in 3 departments, GTV→CTV→ITV→PTV in 8 departments, and GTV→CTV→PTV in 1 department.For 10%-100% patients with LA-NSCLC, the radiation field needed to be replanned when 38-50 Gy was completed.There was no unified standard for the optimal standardized uptake value (SUV) of positron emission tomography (PET)-computed tomography (CT) simulation and delineation.Seven departments had applied magnetic resonance imaging (MRI) simulation and 10 departments had applied stereotactic body radiation therapy (SBRT) for the treatment of early-stage NSCLC.For the delineation of PTV for early-stage NSCLC (T1-2N0M0), GTV→IGTV→PTV was performed in 5 departments, IGTV→PTV in 3 departments, and GTV→CTV→ITV→PTV in 2 departments.In all the 12 departments, peripheral early-stage NSCLC was given 6.0-12.5 Gy/fraction, 3-12 fractions and central early-stage NSCLC was given 4.6-10.0 Gy/fraction, 5-10 fractions.The results of discussion on the delineation of target volume for the patient were as follows:respiratory movements should be measured by 4D-CT or simulator;the lung window width/level is 1600/-600 HU and the mediastinal window width/level is 400/20 HU;the primary controversy is whether the involved-field irradiation or elective nodal irradiation should be used for the delineation of CTVnd in the mediastinal lymph node.Conclusions Basic consensus is reached for the delineation of target volume in LANSCLC in these aspects:lung window width/level, respiratory movements and set-up error, primary lesion delineation, the radiation dose in CCRT, and the optimal time for replanning the radiation field.There are controversies on the optimal SUV in the delineation of target volume based on PET-CT simulation, the optimal dose fractionation in SBRT for early-stage NSCLC, and the delineation of CTVnd.

Objective To explore chromobox protein homolog 2 (CBX2) expressions in relation to clinical features of patients and elucidate its role in the progression of hepatocellular carcinoma.Methods Using the Cancer Genome Atlas (TCGA) database,R language was used to analyze the distribution of differentially expressed mRNA in hepatocellular carcinoma.The different expression of CBX2 in HCC and adjacent tissues and its relationship with survival and clinical characteristics of patients were further analyzed.The expression of CBX2 in liver tissues,liver cancer tissue,and L02,HepG2 and SMMC-7721 cell lines was detected by real time-PCR and western blot.The expression of CBX2 was interfered by siRNA in hepatoma cell line.MTT,colony formation,transwell assays,and flow cytometry were used to identify the proliferation,apoptosis,invasion and clone-formation ability of HepG2 and SMMC-7721 cells after CBX2 down-regulation.According to the different data,t-test,ANOVA,chi-square test,and COX regression model were used for statistical analysis.Survival curve was plotted through Kaplan-Meier method.Results TCGA public database analysis showed that the expression of CBX2 mRNA in hepatocellular carcinoma tissues (7.296 ± 1.6115) was significantly higher than normal liver tissues (4.706 ± 0.940) (P =0.000).In addition,the overall survival time of patients with low CBX2 mRNA expression was significantly longer than that of patients with high CBX2 mRNA expression [(5.971 ± 0.411) years vs.(4.650 ± 0.503) years,P =0.001].The expression level of CBX2 mRNA was correlated with the pathological TNM stage (P =0.025) and differentiation degree (P ＜ 0.001) of liver cancer.COX regression analysis showed that CBX2 mRNA expression was an independent predictor of patient survival (P =0.013).siRNA was transfected and compared with the blank control group.The transgenic ability of HepG2 and SMMC-77221 cells decreased significantly at 72h (P ＜ 0.05) and 96h (P ＜ 0.05),and the apoptosis rate (11.430％ ± 0.215％) was higher than blank control group (6.6 00％ ± 0.170％) (P =0.003).The number of invasive cells ((both P ＜ 0.05) and relative colony forming cells ((both P ＜ 0.001) were significantly decreased.In 20 cases of tissue samples,the expression of CBX2 protein (relative expression level 3.020 ±0.269) in liver cancer was higher than that in adjacent tissues (relative expression level 0.886±0.065) (P ＜ 0.001).The overall survival time of patients with low CBX2 expression in liver cancer was longer than that of patients with high expression [(3.670 + 0.576) years vs.(0.834 + 0.153) years,P =0.004].Conclusion An evident high expression of CBX2 is an independent poor prognostic factor in hepatoma.Down-regulation of CBX2 expression can inhibit the progression of liver cancer.Therefore,CBX2 may be a prognostic biomarker and a new target for HCC treatment.

Background and objective Twice-daily radiation concurrent with chemotherapy is one of the standard methods for limited-stage small cell lung cancer. hTe study was to evaluate the feasibility of chemotherapy concurrent with dose-escalating twice-daily radiotherapy by simultaneous integrated boost intensity-modulated radiation therapy (SIB-IMRT) approach in patients with limited-stage small cell lung cancer. Methods Patients with limited-stage small cell lung cancer were included, treated with twice-daily radiotherapy by SIB-IMRT concurrent with chemotherapy of etoposide plus cisplatin. Dose escalation was conducted by“classical”3+3 methods with three patients enrolled in each dose level. The therapeutic gross tumor volume (GTV) was treated according to three consecutive dose levels i.e., 45 Gy at 1.5 Gy twice daily, 50 Gy at 1.67 Gy twice daily and 54 Gy at 1.8 Gy twice daily. hTe planning target volume (PTV) received a dose of 45 Gy delivered in 30 fractions of 1.5 Gy. hTe primary endpoints were acute toxicities. hTe secondary endpoints included overall survival (OS), pro-gression-free survival (PFS) and loco-regional failure-free survival (LRFFS) at 1-year of follow-up. Results Twenty men and six women were included. hTe median age was 52 (30-68) months. 12 patients experienced grade 2 acute esophagitis, and 1 patient developed grade 3 acute esophagitis. Only 3 patients developed Grade 2 pneumonitis. Grade 3 or higher radiation-related pneumonia was not observed. None died of treatment-related causes. With median follow-up of 11.2 months (3.2-36.2 months), 1-year OS, PFS and LRFFS were 89.0%, 51.0%and 85.0%, respectively. Conclusion Dose escalation for twice-daily radiation concurrent with chemotherapy in LS-SCLC has been safely achieved up to 54 Gy for GTV using SIB-IMRT technique.

Background and objective Concurrent twice-daily radiotherapy with chemotherapy of EP regimen is one of the current standard treatments for limited-stage small cell lung cancer. However, the safely tolerated dose of standard chemotherapy for Chinese patients is not decided. hTis study was to evaluate the toxicity and the maximum tolerated dose (MTD) of etoposide and cisplatin concurrent with thoracic radiation therapy for patients with limited-stage small cell lung cancer. Methods Patients with histologically proven limited-stage small cell lung cancer (LS-SCLC) were eligible. hTe pa-tients underwent thoracic radiotherapy (45 Gy, 1.5 Gy bid, 30 fractions for 3 weeks) delivered concurrently with etoposide (100 mg/m2 iv, days 1-3) and cisplatin dose escalating from the two levels ( 70 mg/m2 and 75 mg/m2 on d1). hTe primary end-points were hematologic toxicities during treatment. hTe secondary endpoints were non-hematologic toxicities, overall survival (OS) and progression-free survival (PFS). According to Common Terminology Criteria for Adverse Events 4.0 (CTC-AE 4.0), maximum tolerant dosage (MTD) was deifned as the highest safely tolerated dose at which no more than one patient out of six experiences dose-limiting toxicity (Grades 4 hematologic), with the next higher dose having at least two out of six patients experience dose-limiting toxicity. Results From January 2013 to August 2016, 20 patients were enrolled in this study. hTe median age was 49.5 (30-68). Atfer the ifrst 6 patients were enrolled in Arm 1 (70 mg/m2 on d1), one patient had Grade 4 neu-tropenia. Another 14 patients were enrolled in Arm 2 (75 mg/m2 on d1), one patient had Grade 4 neutropenia. hTe MTD was determined to be etoposide (100 mg/m2 iv, d1-d3) and cisplatin dose (75 mg/m2 on d1). 4 patients had≥Grade 3 neutropenia and 1 patients had≥Grade 3 acute esophagitis in Arm 1. 10 patients had≥Grade 3 neutropenia and no patient had≥Grade 3 acute esophagitis in Arm 2. All patients with a median follow-up time was 9.0 months, median OS and PFS were not achieved, 1-year OS and PFS were 91%and 61%, respectively. Conclusion hTe MTD of RT with concurrent chemotherapy of EP regi-men for patients with LS-SCLC was etoposide (100 mg/m2 iv, d1-d3) and cisplatin dose (75 mg/m2 on d1).

The 2013 National Practitioners Qualification Examination Work Conference proposed to reform the traditional one-stop test into two stage. In May 2017, Fujian University of Traditional Chinese Medicine participated in this empirical study as a pilot for the first time. This paper made a preliminary summary and analysis of the results of the phased examination. Because of the absence and deficiency of clinical skills of students in our school, we carried out a series of reform measures for clinical skill courses: enhancing teaching facilities, enriching teaching contents and methods, reforming assessment mode and strengthening the monitoring of the teaching quality of clinical skills. According to the students, reform measures are conducive to consolidating theoretical knowledge and improving clinical skills, laying a good foundation for successfully passing the national examination of medical practitioners and becoming a qualified medical talent.