Objective To set up an antibody-capture ELISA method to detect the Japanese encephalitis virus(JEV)antibody.Methods ELISA plate was coated with the monoclonal antibody which was specific to the envelope protein epitope E39 of JEV,JEV SA14-14-2 strain as the source of antigen was used to absorb the monoclonal antibody,the absorbed virus used to capture the JEV'S antibody.The antibody that captured ELISA was established.The indirect ELISA method using the virus particles from cell culture was compared with coating ELISA plate,105 clinical serum were checked.Results The background in indirect ELISA assay could not be abscised,positive and negative serum diluted in a ratio of 1:10,1:100,1:1000,the relative value of A posative/A negative were 1.02,0.99,1.13,all<2.1.But the antibody-captured ELISA method when the serum dilution was 1:10,1:100,the A posative/A negative were 3.57,2.94,all>2.1;when the dilution was 1:1000,the A posative/A negative was 1.42,<2.1,it meant the method could distinguish the positive and negative serum efficiently when the dilution Was 1:100,the background problem in indirect ELISA assay could be solved.Antibody-capture method was used to check 105 serum samples,the A posative/A negative over a range of 0.257～0.321(0.262±0.050),all<2.1,no positive sample found.Conclusion The antibody-capture ELISA method has been preliminary set up with a high specificity,capable of quickly identifying JEV from other virus.

Objective To explore economic valuation of the minimally invasive esophagectomy.Methods The cases undergone esophagectomy were studied retrospectively.Monofactorial analysis was done to evaluate the impact of various factors on the overall hospitalization costs.The correlated factors then were used as independent variables to carry out the multiple linear regression aimed at the hospitalization costs.Results The results of monofactorial analysis showed that the accompanying diseases,days during hospitalization,modes of esophagectomy,postoperative complications were correlated factors of the overall hospitalization costs.According to the multiple linear regression,it were sorted based on descending order duing to its effect of largenning the hospitalization costs:postoperative complications (β =0.439,P =0.000),days during hospitalization (β =0.397,P =0.000),accompanying diseases(β =0.257,P =0.000),modes of esophagectomy(β =0.132,P =0.000).The expenses caused immediately by the operations were statistically more of the minimally invasive surgery than that of the open procedures whatever the numerous or proportion of the overall costs (P =0.000).Conclusion Combined thoracoscopic and laparoscopic esophagectomy was more expensive than open operation.Nevertheless,impact of the other factors,such as postoperative complications,were more significant than that of the different procedure.h was a strategy to minimally invasive osophagectomy that hospitalization costs could be reduced by means of controlling complications and diminishing hospitalization days.

Objective The aim of this study was to investigate the diagnostic value of the intravoxel incoherent motion (IVIM) for discriminating nonmetastatic from metastatic mediastinal lymph nodes in lung cancer.Methods IVIM was performed preoperatively on 66 patients with lung cancer during October 2015 to June 2016 in Capital Medical University Affiliated Beijing Chaoyang Hospital.Preoperative examination including the chest MRI scan,chest CT,head MRI,bone scan-ning and the cardiopulmonary function.None of the patients was undergone any kind of treatment about the cancer before the examinations,the neoadjuvant chemotherapy or radiation therapy,immunotherapy and gene targeted therapy were included.Measured the short axis diameter and the IVIM parameters of the mediastinal lymph nodes,apparent diffusion coefficient(ADC),diffusion coefficient(D),pseudo-diffusion coefficient(D *),and perfusion fraction(f).All the patients underwent the lobectomy or wedge resection and lymph node dissection or systematic sampling.By comparing the IVIM parameters of the nonmetastatic and metastatic mediastinal lymph nodes according to the pathological examination,draw the ROC curves to find the best cutoff value for diagnosis.And 42 cases were used to comparing the value of the MRI and CT in differential diagnosis.Results MRI measured 184 groups mediastinal lymph nodes,including 164 groups without metastasis,20 groups with metastasis.The ADC,D,D *,f values and short axis diameter of the nonmetastatic lymph nodes(n =164) were(2.880 ± 0.785) × 10-3 mm2/s,(0.670 ± 0.179) × 10-3 mm2/s,(0.383 ± 0.123) × 10-3 mm2/s,0.422 ± 0.119,(6.546 ± 1.932) mm,respectively,and (1.897 ±0.657) × 10-3 mm2/s,(0.472 ±0.210) × 10-3 mm2/s,(0.354 ±0.130) × 10-3 mm2/s,0.412 ±0.090,(7.510 ± 2.773) mm respectively for the metastatic lymph nodes (n =20).The ADC and D value of the nonmetastatic lymph nodes were significantly higher than the metastatic lymph nodes(P ＜0.01).While the other paraneters(D *,f,and short axis diameter) between the two groups did not show significantly different.Optimal cutoff values(area under the curve,sensitivity,and specificity) for distinguishing metastatic from nonmetastatic lymph nodes were as follows:ADC =1.890 × 10-3 mm2/s (0.871,92.7％,80.0％);and D =0.648 × 10-3mm2/s(0.740,70.0％,84.1％).Conclusion IVIM is useful to distinguish metastatic from nonmetastatic lymph nodes in lung cancer.The ADC and the D values are significant higher in metastatic lymph nodes,which more sensitive than the other parameters(D *,f,and short axis diameter).As a result,IVIM can be used in the N-stage diagnosis of lung cancer.

The genetic structure of small-scale landscape groups of Oncomelania hupensis in Songzi City ,Hubei Province was identify in this study .O .hupensis snails were collected from 10 habitats in Songzi City ,of which 6 polymorphic microsat-ellite DNA loci (T6-17 ,P101 ,D11 ,B14 ,T4-33 ,and C22) were carried out with GeneScan .The number of alleles (Na) ,het-erozygosity (H) ,fixation index (FST) of snails in each group ,genetic distance between groups ,and the polymorphic informa-tion content (PIC) were calculated .Cluster analysis was then carried out based on genetic distance ,and hierarchical AMOVA calculation was conducted .By certified the shells of snails ,10 groups were divided into light and ribbed shell (including shallow rib and deep rib) .There were 141 alleles in total detection on 10 populations and 20-34 alleles in each locus ,which were detec-ted for 23 .5 on average .The average number of alleles in 6 loci was 1 .575 and the number of alleles in each locus was uneven , showing large numerical differences ranged from 0 .445 to 3 .060 .The average observed heterozygosity (Ho) ranged from 0 .438 ue between paired populations was from -0 .015 64 to 0 .252 47 ,and the polymorphic information content in the population ranged from 0 .528 -0 .857 ,showing a high polymorphism .Hierarchical AMOVA calculations showed that inter-individual variation of the snails occupied 88 .4% of the total variations .Cluster analysis revealed that the three ribbed shell population in Munu Kou Village ,Hengti Village and Yixing Village first clustered to the three light shell population in Mashizizu Village , Mingzhu Village and Tuqiao Village ,then clustered to the light shell population in Tuanshan Village and Jiama Cao Village with the two shallow rib population in Desheng Village and Tianmu He Village .Under the different landscape environment of Songzi Area ,there were different shells presenting on the morphology of O .hupensis .Although there was a rich diversity on O .hupensis of Songzi City ,the genetic differences mostly present in individuals .Different groups didn’t show the significant genetic differentiation among the different shell morphology of O .hupensis .

Objective To investigate early and midterm outcomes after minimally invasive1-stop or 2-stage MICSCABG and PCI hybrid surgery.Methods Between June 2011 and February 2015, 28 patients(19 males and 9 females) with multiple coronary artery disease underwent MICS-CABG and PCI hybrid surgery, 4 cases of which were1-stop hybrid surgery and 24 cases were 2-stage.All patients were followed up to major adverse events(recurrent angina, secondary myocardial infarction, congestive heart failure, serious arrhythmias, secondary bypass graft) as the end point event.Results All patients were no perioperative deaths, shock, infection and myocardial infarction.ICU stay time was(22.9 ± 12.5) h, hospital stay time was(6.75 ±2.10) days, intubation time was(21.1 ± 17.7) h.Perioperative1-stop and2-stages hybrid surgery had significant statistical differences in ICU stay time and intubation time[(41.8 ±32.9) h vs.(16.5 ± 8.5) h;(40.0 ± 34.2) h vs.(18.8 ± 8.5) h;P ＜ 0.05].Patients were followed up for 1-44 months, averaged (17.9 ± 12.8) months.The postoperative major adverse event rate was 7.14％, one patient died of cerebral hemorrhage after 6 months, and one patient underwent coronary angiography because angina.Conclusion l-stop or 2-stage MICS-CABG and PCI hybrid surgery provides a safe,effective and cosmetic revascularized method to patients with multiple coronary artery disease.Meanwhile,the procedure does not destroy the way of re-operation.The perioperative and early results of follow-up are satisfactory.

Objective To investigate the eftect of VEGF gene transtection on endothelial progenitor cell derived from murine bone marrow.Methods Wistar rat's bone marrow was obtained, mononuclear cell isolated,and endothelial progenitor cells(EPS)were cultured in EGM-2MV.EPCs were identified by immunocytochemistry and electron microscope.EPCs were transfected by liposome mediated pcDNA3.0-hVEGF165.VEGF protein level was determined in the cultural medium supernatant after VEGF transfection by ELISA.Cultural medium supernatant was used to co-culture with ECV304,VEGF protein activity was evaluated by MTT.EPCs expression of vWF,VEGF,FLK-1 was detected by immunocytochemistry.Results EPCs were effectively enriched by EGM-2MV,and the EPCs obtained express the typical cell surface markers such as CD34,CD133,FLK-1.The concentration of VEGF protein in supernatant reaches 1280 pg/ml in the 7th day after pcDNA3.0-hVEGF transfection.No influence of EPCs proliferation could be found after transfeetion.The cell surface marker expression of VEGF,FLK-1, vWF became higher with time,and the ratios of positive cell were 88.52%,82.65% and 95.97% respectively.Conclusions pcDNA3.0-hVEGF165 transfeet EPCS mediated by liposome could excrete a high concentration of functional VEGF protein.It is helpful for EPC to maintain the characters of endothelial cell after VEGF gene transfection and differentiate to mature endothelial cell.

Objective To explore the causes of congenital hypothyroidism by single photon emission computed tomography(SPECT) thyroid imaging.Methods Nineteen cases with congenital hypothyroidism were collected,and SPECT thyroid imaging was performed 15 minutes late after injected 99m TcO_ 4- 18.5-37.0 MBq.Results Five cases were found with normal glands,1 case with enlarged thyroid, 2 cases with enlarged thyroid and increased radiation,4 cases with ectopic thyroid,4 with a hypogenetic thyroid,and the last 3 cases showed no thyroid imaging.Conclusions The causes of congenital hypothyroidism can be found by SPECT thyroid imaging,which is important to the diagnosis of congenital hypothyroidism combined with detection of T_3,T_4,TSH.

Objective To verify the accuracy of the mathematical model of gastrointestinal decompression after esophagectomy of esophageal cancer and explore predictive value of the mathematical model in the postoperative complications.Methods The retrospective case-control study was conducted.The clinicopatholo gical data of 192 patients with esophageal cancer who underwent esophagectomy in the Beijing Chaoyang Hospital of Capital Medical University between October 2013 and October 2016 were collected.Among 192 patients,160 didn't have postoperative complications and 32 had postoperative complications (7 with postoperative anastomotic leakage,9 with pulmonary infection and 16 with dysfunction of gastralintestinal tract).Patients selected the appropriate surgical procedures according to individual conditions,and then volume of gastrointestinal decompression was recorded daily.According to the regression equation of influencing factors of volume of postoperative gastrointestinal decompression:average daily drainage volume within 5 days (mL)=262.287 + 132.873 × tubular stomach-72.160 × smoking history-27.904 × pathological type of tumor-36.368 × age,predictive value of postoperative gastrointestinal decompression was calculated and compared with real volume of gastrointestinal decompression.Observation indicators:(1) comparison between predictive value and real volume of postoperative gastrointestinal decompression in patients without complications;(2) comparison between predictive value and real volume of postoperative gastrointestinal decompression in patients with complications.Measurement data with normal distribution were represented as (x)±s and comparison was analyzed using the pairedsamples t test.Measurement data with skewed distribution were described as M (range),and comparison was analyzed using the Wilcoxon signed rank tests.Results (1) Comparison between predictive value and real volume of postoperative gastrointestinal decompression in patients without complications:predictive value and real volume of postoperative gastrointestinal decompression in 160 patients without complications were respectively 187 mL (range,58-392 mL) and 207 mL (range,20-570 mL),with no statistically significant difference (Z=-1.106,P＞0.05).(2) Comparison between predictive value and real volume of postoperative gastrointestinal decompression in patients with complications:7 patients had postoperative anastomotic leakage,including 1 with cervical anastomotic leakage and 6 with chest anastomotic leakage.The predictive value and real volume of postoperative gastrointestinal decompression in 7 patients with anastomotic leakage were respectively (215±58)mL and (338± 106)mL,with a statistically significant difference (t=-3.139,P＜0.05).The predictive value and real volume of postoperative gastrointestinal decompression in 9 patients with postoperative pulmonary infection were respectively (176±61) mL and (239± 111) mL,with no statistically significant difference (t =-1.805,P＞0.05).The predictive value and real volume of postoperative gastrointestinal decompression in 16 patients with dysfunction of gastralintestinal tract were respectively (236 ± 60) mL and (357 ± 107) mL,with a statistically significant difference (t =-4.716,P＜ 0.05).Conclusions The mathematical model of gastrointestinal decompression after esophagectomy of esophageal cancer is correct and feasible.There is a predictive value for patients with postoperative anastomotic leakage and dysfunction of gastralintestinal tract.

Objective To investigate the clinical value of preoperative serum carcinoembryonic antigen (CEA) detection in the prediction of esophageal cancer lymph node metastasis.Methods The clinical data of 111 patients with esophageal cancer who were admitted to the Chaoyang Hospital of Capital Medical University between December 2010 and January 2014 were retrospectively analyzed.Patients received preoperative serum CEA examination and enhanced CT of the chest.The surgical procedures were selected according to the condition of patients, including radical resection of esophageal cancer via left thoracic approach, transabdominal right thoracic approach (open and laparoscopic surgeries), cervico-thoracic-abdominal triple incision (open and laparoscopic surgeries) and transabdominal incision.The international standard was used for tumor location and TNM stage of esophageal cancer.The count data and comparison of ordinal data in the univariate analysis were analyzed using the chi-square test, Fisher exact probability and rank-sum test, respectively.The multivariate analysis was done using the stepwise logistic regression.The ROC curve was used for evaluating diagnostic value of serum CEA examination and enhanced CT of the chest.All the 111 patients were divided into 4 groups according to the interquartile range results of the CEA examination, and the lymph node metastasis rates of 4 groups were compared by the chi-square test.Results All the 111 patients underwent successful radical resection of esophageal cancer after preoperative serum CEA detection and enhanced CT of the chest, including 40 via left thoracic approach, 56 via transabdominal right thoracic approach, 8 via cervico-thoracic-abdominal triple incision and 7 via transabdominal incision.There were 3 patients with upper thoracic esophageal cancer, 52 with middle thoracic esophageal cancer, 36 with lower thoracic esophageal cancer and 20 with cancer of gastro-esophageal junction.The postoperative pathological type included 84 squamous cell carcinomas, 23 adenocarcinomas and 4 other carcinomas.There were 44 patients with negative lymph node metastases and 67 with positive lymph node metastases.The positive rate of elevated serum CEA in the 111 patients was 36.04％ (40/111).Tumor location, pathological type and N stage of tumor were clinical pathological factors affecting the positive rate of serum CEA of patients (Z =6.815, 6.608, 16.928, P ＜0.05).N stage of tumor was an independent risk factor affecting the positive rate of serum CEA of patients by multivariate analysis [OR =2.206, 95％ confidence interval (CI) :1.370-3.552, P ＜ 0.05].The T stage of tumor and serum CEA level were risk factors affecting lymph node metastasis of esophageal cancer by univariate analysis (Z =18.971, x2=10.081, P ＜0.05), and those were also independent risk factors affecting lymph node metastasis of esophageal cancer by multivariate analysis (OR =3.558, 3.936, 95％ CI: 1.798-7.041, 1.480-10.469, P ＜0.05).The lymph node metastasis rates of esophageal cancer were 46.43％, 48.28％ , 55.56％ and 92.59％ when CEA level≤ 1.75 μg/L, 1.75 μg/L ＜ CEA level ≤ 2.68 μg/L, 2.68 μg/L ＜ CEA level ≤4.21 μg/L and CEA level ＞ 4.21 μg/L by the stratified analysis, respectively, with a significant difference among the 4 groups (x2=16.026, P ＜ 0.05).The areas under the curve of CEA level and enhanced CT of the chest for lymph node metastasis were 0.687 (95％ CI: 0.590-0.785) and 0.689 (95％ CI: 0.591-0.788) by ROC curve, which were significantly different from the area under the guides (P ＜0.05).The areas under the curve of CEA level and enhanced CT of the chest for lymph node metastasis were 0.785 (95％ CI: 0.697-0.873, P ＜ 0.05).Conclusions Serum CEA detection not only has certain predictive value for lymph node metastasis of esophageal cancer, but has a higher predictive value combined with enhanced CT of the chest.There is a risk of lymph node metastasis for patients with deep tumor invasion and elevated CEA level, and the range of lymph node dissection should be expanded.

Objective To extract the loci of murine MHC gene and construct plasmids.Methods The RNA of mice was extracted and reversely transcribed into cDNA.By using nested PCR,the products were connected with T vector,cloned,and sequenced.Subsequently,the genes were digested by endonucleases,connected with expression vector,and sequenced again to choose the correct clones.Results After the nested PCR,the products were approved by sequencing.After being connected with the vectors,they were approved again by sequencing and the correct clones were chosen.Conclusion All of the loci of the MHC gene can be obtained by nested PCR.The plasmids from the correct clone can be used in the further experiments of transferring the gene to mitigate the transplantation rejection.