Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

AIM: To analyze the difference of endonuclease domain containing 1 (ENDOD1) expression be-tween benign prostatic hyperplasia ( BPH ) tissues and prostate cancer ( PCa ) tissues and to investigate the effect of ENDOD1 on the biological function of human prostate cancer cells .METHODS: The BPH samples ( n=20 ) and PCa samples (n=21) were processed and analyzed according to the instruction of immunohistochemical (IHC) staining.The mRNA and protein levels of ENDOD 1 in the normal prostate epithelial cells and prostate cancer cells were evaluated by RT -qPCR and Western blot , respectively .The recombinant plasmids pCMV-N-Flag-ENDOD1 was constructed and was trans-fected into the human prostate cancer cells .The proliferation , apoptosis , migration and invasion abilities of the prostate cancer cells were evaluated by MTT assay , flow cytometry, Transwell migration and Matrigel invasion assays , respectively. RESULTS:The analysis of variance of the immunoreactivity score showed that PCa tissues with high Gleason score dis -played significantly lower ENDOD1expression than that with low Gleason score and BPH (P<0.05).The expression of ENDOD1 at mRNA and protein levels in PC3 cells and DU145 cells was significantly lower than that in the LNCap cells (P<0.05).The proliferation of DU145 transfected with ENDOD1 was inhibited.The flow cytometry indicated that ENDOD1 over-expression in the DU145 cells resulted in a notable increase in G0/G1 phase arrest (P<0.05), but the ap-optotic rates showed no statistical difference .The results of Transwell assay showed that migration and invasion abilities of the cells were also inhibited after transfection with over-expressing ENDOD1 plasmid (P<0.05).CONCLUSION: The expression of ENDOD1 significantly decreased in prostate cancer with high Gleaon score .Meanwhile, the ENDOD1 is spe-cifically down-regulated in androgen independent prostate cancer (AIPC) cell lines.Over-expression of ENDOD1 remark-ably inhibits the proliferation , migration and invasion abilities of AIPC .

Objective:To evaluate the efficacy and adverse reaction caused by Capecitabine compared with S-1 as maintenance treat-ments for patients with advanced gastric cancer (AGC) after first-line induction chemotherapy. Methods:A total of 130 AGC patients who did not suffer disease progression after first-line chemotherapies, including XELOX (four to six cycles), SOX (four to six cycles), and mFOLFOX6 regimen (six to eight cycles), were randomized into three groups. The Capecitabine group (Cap) received maintenance che-motherapy with Capecitabine (1 000 mg/m2 twice daily for 14 days, 21 days/cycle), while the S-1 group (S1) received S-1 (40, 50, or 60 mg according to the body surface area and orally administered twice a day for 14 days, 21 days/cycle). The control group was consid-ered as the observation group. Patients with maintenance treatments received drugs until disease progression or observation of intol-erant toxicity. Results:A total of 44, 33, and 53 patients received XELOX, SOX, and mFOLFOX6 regimens, respectively. The overall DCR was 63.1%. Among the 82 patients, 35, 28, and 19 belonged to the Cap, S1, and observation groups, respectively. The comparison be-tween the efficacy of treatments in the Cap and S1 groups did not show statistically significant differences (P=0.678). The median time of progression was 8.5 months in the Cap group and 9.0 months in the S1 group (P>0.05). Both groups showed better responses than the observation group, which demonstrated a median progression of 6.0 months (P<0.001). The median overall survivals were 14.5, 15.0, and 14.0 months in the Cap, S-1, and observation groups, respectively (P=0.188). The most common adverse effects observed among the patients with maintenance treatments included myelo-suppression, gastrointestinal reaction, fatigue, hand-foot syndrome, and stomatitis. No death occurred in relation to the therapy. Conclusion:The effectiveness of Capecitabine and S-1 as maintenance chemotherapies in AGC patients after the first-line induction chemotherapy are similar, and both can prolong the time of disease pro-gression with low toxicity.

Objective:To explore a medical big data algorithm to screen the core indicators in clinical database that can be used to evaluate the prognosis of elderly patients with pneumonia.Methods:Based on the clinical database of a Beijing Chaoyang Hospital Consortium Chaoyang Emergency Ward in Beijing Chaoyang Hospital, Capital Medical University, patients with pulmonary infection were selected through the big data retrieval technology. According to the prognosis at the time of discharge, they were divided into death group and survival group. The general data of patients were collected, including gender, age, blood gas and laboratory indices. A computer language called Python was used to make batch calculations of key indicators that affect mortality in elderly patients with pneumonia. Logistic regression analysis was used to analyze the relationship between laboratory indicators and patients' prognosis. Receiver operating characteristic curve (ROC curve) was drawn to analyze the predictive value of screening method for patients' prognosis.Results:A total of 265 patients were included in the study, 64 died and 201 survived. The data of the first detection indexes of each patient after admission were collected, and 23 key indicators with significant differences were selected from 472 indicators: blood routine indicators ( n = 7), blood gas indicators ( n = 3), tumor markers indicators ( n = 3),coagulation related indicators ( n = 4), and nutrition and organ function indicators ( n = 6). ① The key indicators of blood gas in patients died of pneumonia: Cl - was 97-111 mmol/L in 51.6% (33 cases) of patients, lactic acid (Lac) was 0.5-2.5 mmol/L in 81.2% (52 cases) of patients, and H + was 0-46 mmol/L in 87.5% (56 cases) of patients. ② The key indicators of blood routine of patients died of pneumonia: hemoglobin count (Hb) of 46.9% (30 cases) patients was 80-109 g/L, the eosinophils proportions (EOS%) in 67.2% (43 cases) patients was 0.000-0.009, the lymphocytes proportions (LYM%) in 51.6% (33 cases) patients was 0.00-0.09, the red blood cell count (RBC) in 50.0% (32 cases) patients was (3.0-3.9)×10 12/L, the white blood cell count (WBC) in 54.7% (35 cases) patients was (0.0-9.9)×10 9/L, and the red blood cell volume distribution width coefficientof variability (RDW-CV) in 48.4% (31 cases) patients was 10.0%-14.9%, serum C-reactive protein (CRP) was 0.0-49.9 mg/L in 48.4% (31 cases) patients. ③ The key indicators of tumor markers in patients died of pneumonia: 76.6% (49 cases) of patients had negative free prostate specific antigen/total prostate specific antigen (FPSA/TPSA, the ratio was 0), 92.2% (59 cases) had cytokeratin 19 fragment (CYFRA21-1) between 0.0-11.0 μg/L, and 75.0% (48 cases) had carbohydrate antigen 125 (CA125) between 0-104 kU/L.④ The key coagulation indexes of patients died of pneumonia: 68.8% (44 cases) of patients had activated partial thromboplastin time (APTT) of 57-96 s, 73.4% (47 cases) of patients had D-dimer of 0-6 mg/L, 93.8% (60 cases) of patients had thrombin time (TT) of 14-22 s, and 89.1% (57 cases) of patients had adenosine diphosphate (ADP) inhibition rate of 0%-53%. ⑤ Nutrition and organ function key indicatorsin patients died of pneumonia: 92.2% (59 cases) of brain natriuretic peptide (BNP) in patients with 0, 46.9% (30 cases) of patients had prealbumin (PA) of 71-140 mg/L, 90.6% (58 cases) of the patients with uric acid (UA) for 21-41 μmol/L, 75.0% (48 cases) of the patients with albumin (Alb) to 10-20 g/L, 93.5% (60 cases) of patients had albumin/globulin ratio (A/G ratio) of 0-0.9, 84.4% (54 cases) of the patients with lactate dehydrogenase (LDH) from 0-6.68 μmol/L·s -1·L -1. ⑥ Logistic regression analysis and ROC curve analysis: Logistic regression analysis showed that PA and Lac were the prognostic factors. PA could reduce the risk of death by 0.9%, Lac could increase the risk of death by 69.4%; the area under ROC curve (AUC) between laboratory indicators and the prediction effect of death prediction model for patients' prognosis was 0.80, which showed that the classification effect was better, and this study model could better predict the prognosis of elderly patients with pneumonia. Conclusion:By using big data technology, 23 core indicators for evaluating the prognosis of elderly patients with pneumonia can be screened from the clinical database of emergency ward, which provides a new perspective and method for clinical evaluation of the prognosis of elderly patients with pneumonia.

Objective:To monitor iodized salt consumption and evaluate iodine deficiency status in 2014 in China.Materials and Methods:In 2014,a nationwide cross-sectional survey was conducted in 31 provinces (in this study,provinces,autonomous regions and municipalities in Mainland China were named as provinces).Probability proportional to size sampling method was adopted to recruit children ages 8-10 and pregnant women.47,467 children's and 18,994 pregnant women's urine samples were collected and 47,706 children's thyroid volumes were examined.Iodine content in salt was determined with 46,900 edible salt samples from children's households;urinary iodine concentration (UIC) was tested from children and pregnant women's urine samples;thyroid volume of children was assessed by ultrasound.Results:The national coverage rate of iodized salt and consumption rate of qualified iodized salt were 96.3％ and 91.5％,respectively.Median iodine content in iodized salt was 25.2 mg/kg.In 22 of 31 provinces,the provincial coverage rates of iodized salt were over 95％.And consumption rates of qualified iodized salt were more than 90％ in 21 provinces.In this study,the national median urinary iodine concentration (MUIC) of children in China was 197.9 μg/L.At the provincial level,MUIC of children in 19 provinces was 100-199 μg/L,which in 12 provinces was 200-299 μg/L.The national MUIC of pregnant women in 2014 was 154.6 μg/L,slightly higher than the lower limit of the WHO criteria for adequate (150-249 μg/L).At the provincial level,MUIC of pregnant women in 18 provinces was 100-149 μg/L,which in 13 provinces was 150-249 μg/L.The national prevalence of goiter among children in 2014 was 2.6％,of which only Shandong province (5.6％) exceeded the national standard (5％).Conclusions:In China,iodine deficiency disorders (IDD) has been eliminated since 2005.And in 2014,the IDD-free status still remained.

Objective To investigate the medium- and long-term effects and safty of Pipeline embolization device ( PED) for the treatment of large and giant intracranial anterior circulation aneurysms. Methods From December 2014 to December 2016,the data of 36 consecutive patients with large and giant aneurysm ( 36 large and giant intracranial anterior circulation aneurysms ) treated with PED in Donglei Brain Doctor Group were analyzed retrospectively. The diameter of the aneurysms was 12 -33 mm, (mean16.6±4.5mm),andthenecksizewas4-10mm(mean6.1±1.5mm).Eightaneurysmswere located in the carotid cavernous sinus segment,22 in the ophthalmic artery segment,5 in the internal carotid artery posterior communicating segment, and 1 in the M1 segment of middle cerebral artery. Seven aneurysms were only treated by PED,28 aneurysms were treated by PED in combination with coil embolization,and 1 aneurysm was treated by double PEDs. The modified Rankin scale ( mRS) score was used to evaluate the prognosis of the patients. Results (1) The clinical prognosis of the patients was followed up by telephone and outpatient department for 6-33 months. Twenty-five patients were followed up by DSA,23 aneurysms (92%) were occluded totally (Raymond gradeⅠ) and 2 (8％) were occluded near totally ( Raymond grade Ⅱ) . ( 2 ) Seven patients were treated with PED alone. Four patients were cured totally after 6 months follow-up,1 was occluded subtotally,2 were not cured;6 were cured in the last follow-up (33 months),and the other aneurysm was gradually reduced;17 of 28 patients treated with PED in combination with coils received DSA follow-up. They were followed up for 6-8 months. All the aneurysms were totally occluded ( Raymond grade Ⅰ) . 1 aneurysm was treated by 2 PEDs, DSA revealed micro-aneurysm-like development at 8 months after procedure. The aneurysms were basically occluded after 15-month follow-up. (3) MRI confirmed after operation that 10 patients had asymptomatic scattered spotted ischemic foci,4 had cerebral parenchymal hemorrhage,1 of them died,1 recovered well after treatment (mRS 1),and the other 2 were asymptomatic cerebral hemorrhage. The occupying effect of 24 cases disappeared, 8 had obvious improvement,and 3 did not have any obvious change. Conclusions The occlusion rate of the treatment of large and giant intracranial aneurysms with PED was high. The results of medium-term follow-up showed that the occlusion rate of PED in combination with coils in the treatment of aneurysms was higher than that of PED alone. The long-term follow-up results showed that the occlusion rate of patients treated with PED alone (including one or more) was gradually increased with time. The safety of the surgery needs to be further confirmed by a large sample study.

Objective:This observational study was aimed to analyze the clinical characteristics of infective endocarditis (IE) in patients with hypertrophic cardiomyopathy (HCM).Methods:A total of 668 patients with IE, and 7 427 patients with HCM were treated in Fuwai Hospital from August 2006 to December 2018. Among them, 14 patients were diagnosed with HCM and IE. The clinical characteristics of these patients including clinical manifestations, pathogen distribution, echocardiography features, in-hospital treatment and outcomes were analyzed retrospectively.Results:The proportion of HCM patients with IE was 0.19%,with the estimated incidence of 0.15/1 000 person-years in HCM patients. Of the 14 patients, 11 patients were male. The most common clinical manifestations were fever and heart murmur, and the main complications were heart failure (12/14) and bacterial embolism (8/14). There were 8 cases (8/14) with positive blood culture, and all causative bacteria were gram positive coccus, in which 5/8 were Streptococcus. The median interventricular septum thickness was (21.2±2.7) mm, and left ventricular outflow obstruction was severe based on echocardiography (Echo) examination. The Echo showed that vegetation was found in all 14 patients and most of the vegetation attached at the anterior leaflet of mitral valve (12/14). The proportions of patients with circulatory embolism (8/14) and valve lesions (12/14) were relatively high. Most cases (10/14) were cured, especially those underwent cardiac surgery (8 cases). The rest 4 cases died with 2 in hospital and 2 after auto-discharge. Conclusions:HCM patients complicated with IE are rare. Septic embolization and valve lesions are common in these patients. IE patients with HCM might have a poor prognosis compared to those without HCM and should receive cardiac surgery as early as possible.

Objective: To investigate the clinical value of Bayesian network in predicting survival of patients with advanced gallbladder cancer(GBC)who underwent curative intent surgery. Methods: The clinical data of patients with advanced GBC who underwent curative intent surgery in 9 institutions from January 2010 to December 2015 were analyzed retrospectively.A median survival time model based on a tree augmented naïve Bayes algorithm was established by Bayesia Lab software.The survival time, number of metastatic lymph nodes(NMLN), T stage, pathological grade, margin, jaundice, liver invasion, age, sex and tumor morphology were included in this model.Confusion matrix, the receiver operating characteristic curve and area under the curve were used to evaluate the accuracy of the model.A priori statistical analysis of these 10 variables and a posterior analysis(survival time as the target variable, the remaining factors as the attribute variables)was performed.The importance rankings of each variable was calculated with the polymorphic Birnbaum importance calculation based on the posterior analysis results.The survival probability forecast table was constructed based on the top 4 prognosis factors. The survival curve was drawn by the Kaplan-Meier method, and differences in survival curves were compared using the Log-rank test. Results: A total of 316 patients were enrolled, including 109 males and 207 females.The ratio of male to female was 1.0∶1.9, the age was (62.0±10.8)years.There was 298 cases(94.3%) R0 resection and 18 cases(5.7%) R1 resection.T staging: 287 cases(90.8%) T3 and 29 cases(9.2%) T4.The median survival time(MST) was 23.77 months, and the 1, 3, 5-year survival rates were 67.4%, 40.8%, 32.0%, respectively.For the Bayesian model, the number of correctly predicted cases was 121(≤23.77 months) and 115(>23.77 months) respectively, leading to a 74.86% accuracy of this model.The prior probability of survival time was 0.503 2(≤23.77 months) and 0.496 8(>23.77 months), the importance ranking showed that NMLN(0.366 6), margin(0.350 1), T stage(0.319 2) and pathological grade(0.258 9) were the top 4 prognosis factors influencing the postoperative MST.These four factors were taken as observation variables to get the probability of patients in different survival periods.Basing on these results, a survival prediction score system including NMLN, margin, T stage and pathological grade was designed, the median survival time(month) of 4-9 points were 66.8, 42.4, 26.0, 9.0, 7.5 and 2.3, respectively, there was a statistically significant difference in the different points(P<0.01). Conclusions The survival prediction model of GBC based on Bayesian network has high accuracy.NMLN, margin, T staging and pathological grade are the top 4 risk factors affecting the survival of patients with advanced GBC who underwent curative resection.The survival prediction score system based on these four factors could be used to predict the survival and to guide the decision making of patients with advanced GBC.

Objective To evaluate the efficacy and safety of Willis covered stent in the treatment of traumatic carotid cavernous fistulae (tCCF).Methods The imaging and clinical data of 7 patients with tCCF treated with Willis covered stent in Shanghai Punan Hospital from November 2015 to June 2018 were analyzed retrospectively.Results Seven Willis covered stent were used in 7 patients.Immediately after stent release,angiography showed that the fistulae completely disappeared in 6 cases.One patient had a small amount of endoleak,and there was still a small amount of endoleaks after balloon dilatation.It was not further treated.There were no operative complications.During 3-12 months follow-up,no new neurological deficits were found in all patients.Angiographic follow-up of 2 patients showed that the fistulae completely disappeared without recurrence.The internal carotid arteries were patent,no in-stent stenosis and stent displacement.Conclusions Wills covered stent can be used as a treatment for tCCF.Its efficacy is satisfactory and the safety is good.