To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

BACKGROUND: The effect of the interval between previous Helicobacter pylori (H. pylori) eradication and rescue treatment on therapeutic outcomes remains unknown. The aim of this study was to investigate the association between eradication rates and treatment interval durations in H. pylori infections. METHODS: This prospective observational study was conducted from December 2021 to February 2023 at six tertiary hospitals in Shandong, China. We recruited patients who were positive for H. pylori infection and required rescue treatment. Demographic information, previous times of eradication therapy, last eradication therapy date, and history of antibiotic use data were collected. The patients were divided into four groups based on the rescue treatment interval length: Group A, ≥4 weeks and ≤3 months; Group B, >3 and ≤6 months; Group C, >6 and ≤12 months; and Group D, >12 months. The primary outcome was the eradication rate of H. pylori . Drug compliance and adverse events (AEs) were also assessed. Pearson's χ2 test or Fisher's exact test was used to compare eradication rates between groups. RESULTS: A total of 670 patients were enrolled in this study. The intention-to-treat (ITT) eradication rates were 88.3% (158/179) in Group A, 89.6% (120/134) in Group B, 89.1% (123/138) in Group C, and 87.7% (192/219) in Group D. The per-protocol (PP) eradication rates were 92.9% (156/168) in Group A, 94.5% (120/127) in Group B, 94.5% (121/128) in Group C, and 93.6% (190/203) in Group D. There was no statistically significant difference in the eradication rates between groups in either the ITT ( P = 0.949) or PP analysis ( P = 0.921). No significant differences were observed in the incidence of AEs ( P = 0.934) or drug compliance ( P = 0.849) between groups. CONCLUSION: The interval duration of rescue treatment had no significant effect on H. pylori eradication rates or the incidence of AEs. REGISTRATION ClinicalTrials.gov , NCT05173493.
Humans ; Helicobacter Infections/drug therapy* ; Helicobacter pylori/pathogenicity* ; Male ; Female ; Prospective Studies ; Middle Aged ; Anti-Bacterial Agents/adverse effects* ; Adult ; Aged ; Treatment Outcome ; Proton Pump Inhibitors/therapeutic use*

Humans ; Infant ; Biliary Atresia/diagnosis* ; Choledochal Cyst/diagnosis* ; RNA, Circular ; Diagnosis, Differential

Objective     To investigate the clinical characteristics and risk factors for perioperative lung surgery patients with SARS‐CoV‐2 Omicron variant infection. Methods     The clinical data of patients who underwent lung surgery at the Department of Thoracic Surgery, Renmin Hospital of Wuhan University from December 1, 2022 to January 9, 2023 were retrospectively analyzed. The patients were divided into an infection group and a non-infection group according to whether they were infected with SARS-CoV-2. And the clinical data of two groups were collected and compared. Multiple linear regression analysis was used to explore the risk factors affecting the time of hospitalization. Results     A total of 70 patients were enrolled in this study, including 36 (51.4%) males and 34 (48.6%) females at a median age of 61.0 (49.0, 66.8) years. There were 28 patients in the infection group and 42 patients in the non-infection group. The proportion of preoperative abnormal coagulation function and the risk of postoperative pulmonary infection in perioperative patients infected with SARS-CoV-2 were higher than those in the non-infection group (P<0.05). Subgroup analysis found that patients with preoperative SARS-CoV-2 infection were more likely to have pulmonary infection after surgery, but did not prolong the time of hospitalization or increase the risk of severe disease rate. The patients with postoperative SARS-CoV-2 infection had worse clinical prognosis, including longer time of hospitalization (P=0.004), higher ICU admission rate (P=0.000), higher lung infection rate (P=0.003) and respiratory failure rate (P=0.000). Multiple linear regression analysis showed that gender and extent of surgery were independent risk factors for prolonged hospitalization time. Conclusion     Preoperative infection with SARS-CoV-2 Omicron variant will increase the risk of pulmonary infection, but it will not affect the clinical prognosis. However, postoperative infection with SARS-CoV-2 Omicron variant will still prolong the time of hospitalization, increase the ICU rate, and the risk of pulmonary complications.

【Objective】 To develop a simple and practical diagnostic protocol to optimize patient screening for fractional flow reserve（FFR）. 【Methods】 Consecutive patients who underwent both invasive coronary angiography and lesion-specific FFR measurement from July 2013 to August 2018 were retrospectively screened， resulting in a total of 372 patients（390 lesions） for inclusion. Visual estimation（VE） of stenosis was obtained from experts while percent diameter stenosis（DS%）， percent area stenosis（AS%）， lesion length（LL）， minimal lumen diameter（MLD）， and the ratio of LL to the fourth power of MLD（LL/MLD⁴） were recorded by quantitative software. An FFR value of ≤0.80 was considered to indicate the physiological significance of stenosis. 【Results】 The median age（25^th-75^th percentiles） of the included patients was 66 years（59-74 years） and positive FFR results were identified in 77 lesions（19.7%）. The area under the receiver-operating characteristic curve was revealed as 0.711（95% CI 0.663-0.755） for VE， significantly greater than DS%（0.605）， AS%（0.608）， and LL（0.612； P<0.05 for all）， but without significant difference from that of MLD（0.667） and LL/MLD⁴（0.702）. The combination of VE with LL/MLD⁴ yielded a high sensitivity of 89.6%（95% confidence interval ［CI］， 80.6-95.4） and a negative predictive value of 94.4%（95% CI， 89.2-97.5）. 【Conclusion】 Overall， the combination of VE with LL/MLD⁴ can effectively identify those low-risk lesions for ischemia to avoid unnecessary FFR measurement， thus optimizing the use of pressure wire as well as reducing total medical expenditure and potential complications.

Objective To observe the effects of dexmedetomidine on intraoperative wake-up test in Brucellar spondylitis (BS) patients undergoing surgical operation.Methods Using the case control method,thirty-two patients undergoing BS surgical operation from January 2014 to December 2017 in Harbin the Fifth Hospital were enrolled in this study,the patients were randomly classified into the experimental group (n =16) and the control group (n =16).The two groups were anesthetized with midazolam,propofol,sufentanil and cisatracurium,then anesthesia was maintained with sevoffurane inhalation and a continuous intravenous infusion of remifentanil.In the experimental group,dexmedetomidine 0.4 μg· kg-1·h-1 was administered after tracheal intubation,while equal volume saline solution was given to control group.When the wake-up test was performed,the values of mean arterial pressure (MAP),heart rate (HR) and bispectral index (BIS) were recorded at the time points of preinduction (T0),just before wake-up (T1) and awakening (T2).The wake-up time,the amount of bleeding during the wake-up period,the wake-up quality rating and the sedation score were recorded.Results There was significant difference in HR and MAP at T0,T1 versus at T2 in control group [(98.8 ± 21.0) time/min vs (84.5 ± 8.1),(81.8 ± 1.7) time/min,(90.2 ± 7.5) mmHg vs (76.2 ± 5.7),(74.6 ± 8.5) mmHg,1 mmHg =0.133 kPa,P ＜ 0.05].In experimental group,HR and MAP were lower than those in control group at T2,and the difference between the two groups was statistically significant [(86.3 ± 12.3) time/min vs (98.8 ± 21.0) time/min,(77.9 ± 6.3) mmHg vs (90.2 ± 7.5) mmHg,t =-2.901,-4.995,P ＜ 0.05).The wake-up test quality was significantly better in test group than that in control group,the difference was statistically significant (excellent:13 cases vs 4 cases,good:2 cases vs 6 cases,poor:1 case vs 6 cases,x2 =4.571,P ＜ 0.05).The wake-up time and the amount of bleeding during wake-up period were less than that in control group,the difference was statistically significant [(14.5 ± 3.6) min vs (26.1 ± 4.5) min,(239.8 ± 53.9) ml vs (317.3 ± 54.8) ml,t =-7.980,-4.032,P ＜ 0.05].Conclusion Dexmedetomidine when continuous pumped at a rate of 0.4 μg· kg-1· h-1 could reduce the hemodynamic stress response during the wake-up test,improve the wakeup test quality,shorten the wake-up time and effectively improve the safety factor of operation during Brucellar surgical operation.

Objective:BALB/C mice model infected with different Dengue Type 2 virus (DV 2) strains was evaluated and dynamic patterns of humoral immune responses caused by the various viral strains were investigated.Methods:The BALB/C mice were multisubcutaneouly injected by different DV 2 strains. The dynamic changes of the anti-DV 2 IgM/IgG antibody were observed in the infected BALB/C mice by the indrect ELISA and the viremia was observed.Results:The productions of specific antibodies in the infected BALB/C mice after primary and secondary injection with different DV 2 strains showed some difference. Howerer, BALB/C mice infected by the DV 2 B strain from a DHF patient showed the prolonged viremia.Conclusion:The dynamic curve of specific antibody and the viremia induced by various DV 2 strain (DV 2 NGC strain.V.DV 2 B strain) were different in some existence.

Objective:BALB/C mice model infected with DV2 NGC strain was evaluated and dynamic patterns of cytokines were investigated.Methods:The dynamic changes of cytokines were observed in the infected BALB/C mice model by indirect ELISA was observed.Results:The dynamic levels of IL-4、IFN-? in the plasma of BALB/C mice after primary and secondary infected DV2 NGC strain were different.The results showed that the levels of IL-4 were significantly increasing in the early primary infected BALB/C mice while that of IFN-? were lower;In secondary infection ,IL-4 reached a peak on the first day( 4294 668 ? 349 038 pg/ml) and then the levels of IFN-? were higher on the forth and eleventh day.The concentrations of the cytokines produced infectious animals had closely relationship with the doses of DV2.Conclusion:Infection eliciting a dominant humonral immune response induced a high expression of Th2-related cytokines,whereas those show some appearance of Th1 cytokines.Th1 response,as described late for the disease infected with DV2 NGC strain,played a role while it inhibited Th2 response to some extend.Thus,it was responsible to recovery.Th cell and the cytokines were critic to the DV infective diseases.