Objective:To investigate the clinical features and prognosis of septic shock(SS) children with different basic diseases in pediatric intensive care unit (PICU).Methods:The medical records of SS children admitted to PICU at Beijing Children′s Hospital from January 1, 2017 to December 31, 2019 were collected retrospectively.They were grouped according to the presence or absence of basic diseases and types of basic diseases.The clinical characteristics, prognosis and pathogens of SS under different basic diseases were summarized.Results:A total of 218 children with SS were included during the study period, and the overall case fatality rate was 21.6%(47/218). There were 141 cases with basic diseases, accounting for 64.7%(141/218) and 24.1%(34/141) case fatality rate.The mortality rate was highest(37.5%, 17/45) in the malignant hematologic diseases and tumors patients with post-chemotherapy bone marrow suppression, and lowest(16.9%, 13/77) in patients with no underlying diseases.Gram-negative bacterial infection was more common in SS children with underlying diseases(63.1%, 41/65), and was highest in the malignant hematologic diseases and tumors patients with post-chemotherapy bone marrow suppression(80.0%, 20/25). Gram-positive bacteria accounted for the highest proportion in the group without underlying disease(52.1%, 25/48). The incidence of multiple organ dysfunction syndrome(MODS) was the highest(95.6%, 43/45) in the malignant hematologic diseases and tumors patients with post-chemotherapy bone marrow suppression, and the lowest(59.7%, 46/77) in the group without underlying disease.Conclusion:Gram-negative bacteria is the most common pathogen in SS children with underlying diseases, especially in malignant hematologic diseases and tumors patients with post-chemotherapy bone marrow suppression, and with high mortality and incidence of MODS.Gram-positive bacteria is the most common pathogen for those without underlying diseases, with a relatively low mortality and incidence of MODS.

Objective To find out the association between the indicators(pulse concussion lung function test index) of bronchial hyperresponsiveness (BHR) with fractional concentration of exhaled nitric oxide (FeNO) at different control periods among preschool asthmatic children.Methods Totally 74 asthmatic children in the pediatric department of our hospital from April 2015 to February 2017 were enrolled in this study,and 25 children undergoing the lung function and FeNO examination served as the controls,aged 3-5 years old.The cases were divided into three groups according to the standard in 2016 version of the Prevention and Treament Guide of Children Bronchial Asthma:asthma control group(n =26),asthma non-control;group(n =48) and control group (n=25).All data of FeNO,resistance of the respiratory system at 5 Hz(R5),resistance of the respiratory system at 5 Hz (R20),difference of R5 and R20(R5-20),reactance area (AX),reactance of the respiratory system at 5 Hz (X5) and resonant frequency of reactance (Fres) were collected.The FeNO,pulse concussion lung function test value and their association were analyzed.Results (1) The FeNO value of asthma the non-control group was significantly higher than that of the asthma control group and the control group,which were 34.00 ± 18.17,20.23± 11.07 and 28.00± 17.30 respectively.The AX detection value of the asthma non-control group was significantly higher than that of the control group(37.29 ± 15.27 vs.30.17 ± 9.50,P＜0.05).(2)R20 had weak correlation with FeNO in the control group(P＜0.05),while R20 had no correlation with FeNO in the non-control group and control group (P＞0.05).FeNO had no obvious correlation with R5,R520,AX,X5 and Fres in the asthma non-control group,asthma control group and control group(P＞0.05).Conclusion In preschool children with asthma,FeNO can reflect the airway eosinophilic inflammation control,and does not reflect the airway hyperresponsiveness.Thereforeit ie needed to combined with FeNO and IOS indicators (airway hyperresponsiveness index AX,etc.),which can more precisely judge whether asthma being controlled.

Objective To evaluate the effect of oxycodone on function of GABAA receptors in dor-sal root ganglion ( DRG ) neurons of rats with neuropathic pain ( NP ) . Methods Thirty-six adult male Sprague-Dawley rats, weighing 180-220 g, aged 10 weeks, were allocated into 3 groups ( n=12 each) u-sing a random number table method: sham operation group ( group S ) , group NP and oxycodone group ( group O) . The sciatic nerve was only isolated but not ligated in group S. NP was induced by chronic con-striction injury. The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 chromic catgut. Oxycodone 15μg∕kg was intraperitoneally injected once a day for 14 con-secutive days from ligating the sciatic nerve to satisfaction in group O. The thermal paw withdrawal latency( TWL) was measured at 1 day before establishing the model ( T0 ) and 3, 5, 7, 10 and 14 days after es-tablishing the model ( T1-5 ) . The rats were sacrificed after measurement of pain threshold at T5 , and DRG neurons were acutely isolated for recording the amplitude of GABAA receptors-activated currents using whole-cell patch-clamp technique. Results Compared with group S, the TWL was significantly shortened at T1-5, and the amplitude of GABAA receptors-activated currents in DRG neurons was decreased in NP and O groups (P<0. 05). Compared with group NP, the TWL was significantly prolonged at T1-5, and the ampli-tude of GABAA receptors-activated currents in DRG neurons was increased in group O ( P<0. 05) . Conclu-sion Oxycodone can enhance the function of GABAA receptors-activated currents in DRG neurons and thus enhance GABAA receptors-mediated presynaptic inhibition, which may be related to the mechanism of oxyc-odone-induced reduction of NP in rats.

Objective:To investigate the efficacy and safety of polatuzumab vedotin (pola) in treatment of patients with relapsed/refractory diffuse large B-cell lymphoma (DLBCL).Methods:The clinical data of 1 DLBCL patient receiving multiple treatments in Jiangsu Cancer Hospital in May 2016 were retrospectively analyzed, and the related literature was reviewed.Results:The patient, a 57-year-old male, was diagnosed with DLBCL in May 2016. Since June 2016, he had received treatments with four lines including anti-CD20 monoclonal antibody combined with chemotherapy, chemotherapy only and chimeric antigen receptor T cell (CAR-T). However, the disease relapsed or progressed after all treatments. Therefore, the patient had received 6 cycles of pola combined with rituximab since December 2019. Unexpected adverse events were not found during the treatment. The evaluation of clinical efficacy was complete remission after the end of treatment. The progression-free survival time was more than 13 months with follow-up until January 2021.Conclusion:Pola initially shows good efficacy and safety in treatment of patients with relapsed/refractory DLBCL.

Objective To explore the relationship between the serum levels of absent in melanoma 2(AIM2)and apolipoprotein J(Apo J)and the severity and prognosis of vascular dementia(VD).Methods From October 2020 to September 2022,128 patients with vascular dementia were collected as the study group in the Second People's Hospital of Binzhou.Based on the mini-mental state examination(MMSE)score,the severity of the condition was evaluated and the study group was separated into mild group(n=43),moderate group(n=54)and severe group(n=31).After 90 days of treatment,the prognosis of patients was evaluated based on the ability of daily living(ADL)scale score and grouped into grade Ⅰ group(n=66),grade Ⅱ group(n=40)and grade Ⅲgroup(n=22).Another 96 healthy individuals who underwent physical examination were selected as the control group.The expression levels of serum AIM2 and Apo J were detected by qRT-PCR and ELISA methods,and the differences between the groups were compared.Spearman method was applied to analyze the correlation between serum AIM2 and ApoJ expression levels and MMSE scores,and the relationship between serum AIM2 and Apo J expression levels and prognosis in patients with vascular dementia.Logistic regression was applied to analyze the prognostic factors of vascular dementia patients.Results The expression level of serum AIM2 mRNA(3.11±0.57 vs 1.06±0.23)in the study group was higher than those in the control group,while the expression level of serum ApoJ(68.83±12.24 ng/L vs 81.07±13.15 ng/L)was lower than that in the control group,with significant differences(t=32.054,7.174,all P＜0.001).The expression level of serum AIM2 mRNA in the severe group was higher than that in the mild and moderate groups(q=12.807,15.780),the expression level of serum Apo J and MMSE score in the severe group were lower than those in the mild and moderate groups(q=26.201,4.301;12.193,20.802),the expression level of serum AIM2 mRNA in the moderate group was higher than that in the mild group(q=14.688),and the expression level of serum Apo J and MMSE score in the moderate group were lower than those in the mild group(q=20.338,37.537),with significant differences(all P＜0.001).The expression level of serum AIM2 mRNA(5.27±0.60)in the grade Ⅲgroup of vascular dementia was higher than that in the grade Ⅱ(3.36±0.58)and grade Ⅰ groups(2.23±0.55),while the expression level of serum Apo J(51.22±12.21 ng/L)was lower than that in the grade Ⅱ(64.15±12.23 ng/L)and grade Ⅰgroups(77.53±12.25 ng/L).The expression level of serum AIM2 mRNA in the grade Ⅱ group was higher than that in the gradeⅠ group,while the expression level of serum Apo J in the grade Ⅱ group was lower than that in the grade Ⅰ group,with significant differences(q=5.630～30.740,all P＜0.001).The level of serum AIM2 in patients with vascular dementia was negatively correlated with MMSE score but positively correlated with prognosis(ADL grade)(r=-0.535,0.432,all P＜0.001),while the level of serum Apo J in patients with vascular dementia was positively correlated with MMSE score but negatively correlated with prognosis(ADL grade)(r=0.467,-0.496,all P＜0.001).Serum AIM2 mRNA[OR(95％CI):2.746(1.481～5.091)],Apo J[OR(95％CI):0.496(0.311～0.791)],and MMSE score[OR(95％CI):0.568(0.347～0.931)]were influencing factors for the prognosis of vascular dementia patients(all P＜0.05).Conclusion The increase of serum AIM2 mRNA expression level and the decrease of serum Apo J expression level in patients with vascular dementia were closely related to the clinical severity and prognosis of patients,which may effectively evaluate the prognosis of patients.

Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine

Objective:To establish an irritant contact dermatitis(ICD)model induced by sodium dodecyl sulfate(SDS)in mice,and explore its endotype to provide an experimental and theoretical basis for subsequent precise treatment.Methods:Mice were randomly divided into two groups(model group and control group),4%SDS was topically applied to induce ICD in mice,saline was used on control group,the dose and frequency were consistent with model group,and the skin lesions of mice were observed.Epidermal thickness and inflammatory cell infiltration were analyzed by HE staining,toluidine blue staining and immunofluorescence staining.Real-time quantitative PCR was performed to investigate mRNA expression levels of cytokines.Results:Compared with control group,mice in ICD model group showed epidermal thickness on the back of neck,and the numbers of inflammatory cells were increased in dermis.The number of neutrophils,macrophages and T cells were increased.Expressions of Il17a and Il17f mRNA levels were increased.Conclusion:SDS-induced ICD model is successfully established,with the elevated infiltration of neutrophils,macrophages and T cells,and secretion of type 17 cytokines.

Objective: To prepare peptide minotope-based recombinant diagnostic antigen of Epstein-Barr virus (EBV) infection and evaluate its antigenicity preliminarily. Methods: With Trx at the N-terminal and His tag at the C-terminal, the peptide minotope of EBV (GP125, F1, A2, A3C2) was expressed in Escherichia coli and purified by affinity and anion exchange chromatography (designated 'H58’); based on antigenicity of H58 identified by Western blotting (WB), we constructed and evaluated a novel early diagnostic ELISA for EBV infection. Results: The soluble H58 protein with high concentration (2.8 mg/ml) and purity (99.01) was obtained; WB analysis found that there was an obvious band (28 ×10³) on the NC membrane, using H58 anti-Trx monoclonal antibody or acute-phase sera of EBV infection as the first antibody. With the novel ELISA, 50 positive sera of EBV infection and 50 negative sera were detected, displaying that the grouping of OD value of positive serum (95%CI: 1.233-1.489) and negative serum (95%CI: 0.113-0.159) was different (P<0.05) with the sensitivity 98.0%, specificity 96.0% and kappa value 0.940. Conclusions By E. coli expression and affinity and ion exchange chromatography purification, the peptide minotope-based recombinant diagnostic antigen of EBV infection was obtained with excellent antigenicity, which could be applied for serological detection of EBV infection.