Objective To express glycoprotein H(gH)of pseudorabies virus(PRV)in mammalian cells and detect its immunogenicity.Methods The gH gene fragment of PRV-XiangA strain was amplified by PCR and inserted into mammalian cell expression vector pIRES-neo3 to construct recombinant expression plasmid pIRES-gH,which was transfected to HEK-293F cells and cultured in suspension for 5 d. The cell culture supernatant was identified by Western blot and purified by nickel ion chromatography column. The purified gH was emulsified with ISA 201 VG adjuvant to immunize 8 female ICR mice,and 8 mice in control group were immunized with the same amount of adjuvant,which was strengthened at 5 and 8weeks after the first dose respectively. The blood samples were collected at 4,7 and 10 weeks after the first dose and detected for the titer of specific antibody and neutralizing antibody in serum of mice;The mice were challenged with PRVXiangA strain(1. 5 × 104TCID50)by nasal drops 2 d after the third blood collection,and observed for the morbidity and mortality daily.Results The recombinant expression plasmid pIRES-gH was constructed correctly as identified by sequencing. The gH protein was successfully expressed and modified by glycosylation in mammalian cells with good reactivity,and about 625 μg purified protein was obtained under 100 mL culture volume. After three times of immunization,mice produced high level of specific antibody and showed the effect of neutralizing PRV,and the titer of neutralizing antibody reached 1∶256. In the challenge test,all the mice in control group became ill and died,while half of the mice in gH immunized group did not get sick with a survival rate of 50%.Conclusion PRV gH was successfully expressed in mammalian cells,and its immune protection was confirmed for the first time,which provided experimental basis for the further research and application of gH,and also provided a new idea for the development of PRV subunit vaccine.

Objective:To investigate the relationship between preoperative high-sensitivity C-reactive protein (Hs-CRP) levels and clinical outcomes of patients undergoing off-pump coronary artery bypass graft (OPCABG) surgery.Methods:We prospectively selected 123 patients who received OPCABG at Beijing Anzhen Hospital from January 2019 to October 2019, and collected relevant preoperative and postoperative data. Patients were divided into a normal Hs-CRP group(78 cases) and an elevated Hs-CRP group(45 cases)according to the cutoff value (2 mg/L) of Hs-CRP level. The data of the two groups were compared, and regression analysis was performed on the postoperative data with differences to define independent factors.Results:The leukocyte count in the Hs-CRP group was significantly higher than that in the normal Hs-CRP group[(6.5±1.6)×10 9/ml vs. (7.4±2.1) ×10 9/ml, t=-2.839, P=0.005]. In the elevated Hs-CRP group, proportion of patients with atrial post-CABG atrial fibrillation (38% vs. 19%, χ2=5.100, P=0.024), duration of hospitalization[(21.2±7.1)days vs.(16.0±4.6)days, t=-4.469, P=0.000], hospital costs[(143.1±30.7)×10 3 yuan vs. (123.7±21.8)×10 3 yuan, t=-4.090, P=0.000]were significantly higher than those in the normal Hs-CRP level group. Smoking ( OR=1.660, 95% CI: 1.186-1.993, P=0.031) and Hs-CRP ( OR=1.170, 95% CI: 1.050-1.294, P=0.007) were independent risk factors for post-CABG atrial fibrillation. Hs-CRP ( B=0.436, 95% CI: 0.197-0.675, P=0.000) and left ventricular ejection fraction (LVEF, B=-0.180, 95% CI: -0.289--0.071, P=0.001) were independent influencing factors of duration of hospitalization. Hypertension ( B=-11.256, 95% CI: -20.670--1.842, P=0.020), Hs-CRP( B=1.235, 95% CI: 0.217-2.254, P=0.018) and LVEF ( B=-1.168, 95% CI: -1.634--0.702, P=0.000) were independent influencing factors of hospital costs. Conclusion:The preoperative Hs-CRP level of OPCABG is an independent influencing factor of post-CABG atrial fibrillation, duration of hospitalization and hospital costs. This finding lays the foundation for Hs-CRP combined with other indicators to accurately predict the prognosis of OPCABG and screen high-risk patients.

Objective:To clone human anti-ricin antibodies from large phage antibody library.Methods:Panning for a large phage library against ricin toxin was conducted to select specific antibodies against ricin. The binding activities and specificities were tested by ELISA method. Soluble ScFvs were prepared through infecting E coli. HB2151 with the selected phage antibodies and induction with IPTG. Results:Forty positive clones were obtained after 5 rounds of panning, and 12 clones had specific binding ability to ricin toxin. DNA fingerprinting showed 7 different band patterns indicating 7 different positive clones. DNA sequencing showed that variable regions of these ScFvs belonged to different subgroups.Conclusion:Human anti-ricin antibodies were successfully obtained from large phage antibody library.

This paper established a qualitative and semi-quantitative analysis detection method of the micro-arc oxidation titanium and titanium alloy layer of Na, Si, P, S and other impurity elements by X - ray fluorescence. At the same time, from the angle of technological process, we analyzed the impurity element types and sources, and verify the reliability of this method. The locking plates superficial layer contained P element, the average was 1.01%. The occipital plate superficial layer contained Si element, the average was 1.39%. The advantages of this detection method were convenient and fast, and the sample without pretreatment and so forth. So this detection method was the ideal method of qualitative and semi-quantitative analysis to the micro-arc oxidation of surgical implants.

Objective To investigate the effect of fetoscopic photocoagulation of communicating placental vessels in twin-twin transfusion syndrome(TTTS)(selective or non-selective) on the perinatal outcomes.Methods Six cases of TTTS admitted in our department from Dec.2006 to Jun.2008 underwent fetoscopic photocoagulation of communicating vessels.Under direct real-time sonographic guidance,a 3-mm-diameter fetoscope was percutaneously inserted through the maternal abdominal wall into the amniotic cavity of the recipient twin.A combination of ultrasonographic and fetoscopic vision was used to identify the crossing vessels which were systematically coagulated using Nd:YAG laser fiber or bipolar electrocoagulation.Results All the 6 mothers tolerated the procedure without major complications.Two fetal survival rate was 33.33%.Conclusion Fetoscopic photocoagulation of communicating placental vessels in TTTS can effectively improve perinatal outcomes.

Objective:To investigate the predictive value of preoperative left atrial diameter in postoperative atrial fibrillation (POAF) after coronary artery bypass grafting (CABG) in patients with coronary artery disease (CAD) and its influencing factors.Methods:This study was a prospective observational study. A total of 113 patients with CABG surgery were enrolled in Beijing Anzhen Hospital affiliated to Capital Medical University from January 2020 to December 2020. Preoperative coronary angiography, echocardiography and blood test were finished. The number of graft vessel was counted during the operation. The occurrence of POAF was confirmed by electrocardiogram or electrocardiograph monitoring after surgery. The preoperative baseline data, blood test, perioperative complications and other indicators of patients with or without POAF were compared.Results:According to the inclusion criteria and exclusion criteria, 90 patients undergoing CABG were enrolled in the study[71 males and 19 females, aged from 42 to 75, mean age (62±8)years old]. The incidence of POAF was 27.8%(25 cases). The left atrial diameter[40(36-43)mm vs. 35(33-37)mm, P=0.00]and troponin I[4.76(0.87-13.60)ng/ml vs. 1.48(0.56-4.52)ng/ml, P=0.04] in patients with POAF were significantly higher than that in patients without POAF. Compared to patients without POAF, POAF significantly increased the incidence of stroke[4(16%) vs. 0(0), P=0.01], ICU stay[67(24-96) days vs. 22(19-41) days, P=0.00] and hospital stay[21(19-24) days vs. 16(14-24) days, P=0.05]. Binary logistic regression showed that left atrial diameter was significantly correlated with the occurrence of POAF ( OR=1.9, 95% CI: 1.39-2.79, P<0.001). ROC curve analysis showed that 40 mm of left atrial diameter was a predictor of POAF( AUC=0.82, sensitivity 52%, specificity 100%, P<0.001). Linear regression analysis showed that left atrial diameter was significantly positively correlated with hypersensitive C-reactive protein ( Beta=0.24, 95% CI: 0.00-0.26, P=0.043). Conclusion:POAF significantly increased the incidence of stroke after CABG. Preoperative left atrial diameter is an independent predictor for POAF, which is closely related to the systemic inflammatory response.

The aim of this study is to establish a simple and accurate method for vaccine immune e-valuation of porcine pseudorabies virus.In this research,PRV-gD recombinant protein was ex-pressed from mammalian cell HEK-293F as coating antigen,and then the reaction conditions of gD-iELISA were optimized according to checkerboard titration method.The gD-iELISA was used to detect the antibody levels of 211 clinical pig serum samples and the consistency with the neu-tralizing antibody levels wasanalyzed.The results showed that the antigen coating concentration was 0.90 mg/L;the serum to be detected was diluted 1∶100 and incubated at 37 ℃ for 30 min;goat anti-pig IgG-HRP antibody was diluted 1∶55 000 and incubated at 37 ℃ for 30 min;TMB sub-strate was developed at 37 ℃ for 20 min.The method could detect 1∶6 400 diluted PRV positive serum.The results of CSFV,PRRSV,PCV-2,PEDV and FMDV positive sera were all negative by gD-iELISA,and there was no cross-reaction between the method and the above positive sera.The coincidence rate of gD-iELISA and commercial kits was 95.26％,and the intra-and inter-batch co-efficients of variation were both less than 10％.Correlation analysis showed that the correlation coefficient(r)between gD antibody level and neutralizing antibody level was significantly greater than that of gB antibody level,and the gD antibody level had a good linear relationship with the neutralizing antibody level.The results indicated that gD-iELISA was more suitable for vaccine im-mune evaluation of PRV than gB-iELISA.Therefore,the method will have a good prospect of ap-plication in the immunization control of the PRV.

Objective To explore the correlation between serum UA expression levels and disease activity in patients with idiopathic inflammatory demyelinating disease.Methods To test serum UA level of the observation group（40 MS patients in observation group，27 NMO patients in observation group） and 67 controls，compare the difference of UA expression between observation group and control group；The relationship between serum UA level and sex，age of onset and EDSS score was also analyzed.Results（1） The serum UA expression levels in the MS and NMO group was lower than that in the control group；（2）The UA level of male MS patients in the observation group was higher than that of women，all differences were statistically significant（P<0.05）；There was no significant difference in UA between men and women in the observation group（P>0.05）；（3）There was no correlation between UA value，onset age and EDSS score of MS and NMO patients in observation group（P>0.05）.Conclusions The changes of serum UA play an important role in MS and NMO disease activity and pathological injury；Sex has certain influence on the regulation of ua in MS and pathogenesis；The level of serum UA in MS and NMO patients was not related to age and EDSS score.

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).