Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

Objective:To discuss the suitable concentration of D-galactose(D-gal)and modeling period,and establish its induced aging-related cognitive dysfunction model in the mice,and perform a comprehensive evaluation.Methods:Fifty C57BL/6J mice were randomly divided into control group and 100,200,400,and 800 mg·kg-1 D-gal groups,with 10 mice in each group.The mice in various D-gal groups were subcutaneously injected with the corresponding concentration of D-gal once daily;the mice in control group were injected with an equal volume of normal saline.The body mass and water consumption of the mice in various groups were monitored;forelimb grip strength test and experiment on the ability of pole climbing sports were used to evaluate the motor coordination ability of the mice in various groups;novel object recognition test,Y maze test,and Morris water maze test were used to evaluate the cognitive function of the mice in various groups;HE staining and Nissl staining were used to observe the pathomorphology of brain tissue of the mice in various groups;immunohistochemistry method was used to detect the expression of β-galactosidase(β-gal)protein in brain tissue of the mice in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the mRNA expression levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-18,and IL-4 in hippocampus tissue of the mice in various groups;Western blotting method was used to detect the expression levels of β-gal,p53,and p16 proteins in hippocampus tissue of the mice in various groups.Results:The body mass growth trends of the mice in control group and various D-gal groups were consistent and there was no statistically significant difference(P>0.05),and there was no statistically significant difference in water consumption(P>0.05).After 8 weeks of subcutaneous injection of D-gal,compared with control group,the forelimb grip strength values of the mice in 200 and 400 mg·kg?1 D-gal groups were significantly decreased(P<0.05 or P<0.01);the pole-climbing time of the mice in 200 mg·kg?1 D-gal group was significantly prolonged(P<0.05);the recognition indexes of the mice in 200 and 400 mg·kg?1 D-gal groups were significantly decreased(P<0.01);the spontaneous alternation rate of the mice in 100,200,400,and 800 mg·kg?1 D-gal group was significantly decreased(P<0.05 or P<0.01),the escape latency was significantly increased(P<0.05).Spatial probe test showed that compared with control group,the escape latency of the mice in 200 mg·kg?1 D-gal group was significantly increased(P<0.05).The HE staining and Nissl staining results showed that compared with control group,the hippocampus neurons of the mice in 200 mg·kg-1 D-gal group were arranged disorderly,with obvious nuclear pyknosis,nuclear condensation,and abnormal morphology and structure,and the number of Nissl staining positive cells was significantly decreased.The immunohistochemistry results showed that compared with control group,the β-gal expressions in CA1 region,CA3 region,and cortex region of hippocampus tissue of the mice in 200 mg·kg?1 D-gal group were strongly positive.The RT-qPCR results showed that compared with control group,the expression levels of IL-1β,IL-18,and TNF-α mRNA in hippocampus tissue of the mice in 200 mg·kg?1 D-gal group were significantly increased(P<0.05 or P<0.01),and the expression level of IL-4 mRNA was significantly decreased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of β-gal,p53,and p16 proteins in hippocampus tissue of the mice in 200 mg·kg?1 D-gal group were significantly increased(P<0.05 or P<0.01).Conclusion:The aging-related cognitive dysfunction model in the mice can be established by subcutaneous injection of 200 mg·kg?1 D-gal daily for 8 weeks.

From July 23, 2018, to December 31, 2019, a total of 299 219 children in Shandong Province received the fifth dose of the diphtheria, tetanus, and acellular pertussis combined vaccine (DTaP). Among these recipients, the distribution by age was as follows: 20 children under 2 years old (0.01%), 273 996 children aged 2 years (91.57%), 20 242 children aged 3 years (6.76%), 3 932 children aged 4 years (1.31%), 963 children aged 5 years (0.32%), and 66 children aged 6 years and above (0.02%). In total, 1 972 cases of adverse events following immunization (AEFI) were reported after the administration of the fifth dose of DTaP, resulting in an incidence rate of 659.05 per 100 000 doses. Among these, 1 718 cases were classified as common vaccine reactions, with an incidence rate of 574.16 per 100 000 doses, while 247 cases were identified as rare reactions, yielding an incidence rate of 82.55 per 100 000 doses. The incidence of AEFIs, as well as the rates of common and rare reactions, exhibited a significant increasing trend with the number of doses administered (all P<0.001). Among the rare reactions, there were 10 cases classified as severe, resulting in a reported incidence of 3.34 per 100 000 doses.

From July 23, 2018, to December 31, 2019, a total of 299 219 children in Shandong Province received the fifth dose of the diphtheria, tetanus, and acellular pertussis combined vaccine (DTaP). Among these recipients, the distribution by age was as follows: 20 children under 2 years old (0.01%), 273 996 children aged 2 years (91.57%), 20 242 children aged 3 years (6.76%), 3 932 children aged 4 years (1.31%), 963 children aged 5 years (0.32%), and 66 children aged 6 years and above (0.02%). In total, 1 972 cases of adverse events following immunization (AEFI) were reported after the administration of the fifth dose of DTaP, resulting in an incidence rate of 659.05 per 100 000 doses. Among these, 1 718 cases were classified as common vaccine reactions, with an incidence rate of 574.16 per 100 000 doses, while 247 cases were identified as rare reactions, yielding an incidence rate of 82.55 per 100 000 doses. The incidence of AEFIs, as well as the rates of common and rare reactions, exhibited a significant increasing trend with the number of doses administered (all P<0.001). Among the rare reactions, there were 10 cases classified as severe, resulting in a reported incidence of 3.34 per 100 000 doses.

Objective:To explore the immunological characteristics of peripheral blood and genetic variations of 11 immunodeficiency virus(HIV)-negative children with Talaromyces marneffei(TM) infection, thus enhancing the diagnostic and therapeutic levels of TM infection in children. Methods:Clinical data of 11 HIV-negative children with TM infection who presented to Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from January 2010 to December 2022 were retrospectively analyzed, including clinical characteristics, peripheral immune profile and genetic test results.Results:A total of 11 HIV-negative children with TM infections were recruited, involving 9 males and 2 females with a median age of 19 months.The main clinical manifestations were fever (10/11, 90.91%), cough (10/11, 90.91%) and hepatomegaly (7/11, 63.64%). Common severe complications included acute respiratory distress syndrome (7/11, 63.64%) and septic shock (5/11, 45.45%). Finally, 2 children died.Transient neutropenia occurred in 6 cases (6/11, 54.55%), and lymphocytopenia combined with serum immunoglobulin (Ig) G decrease was observed in 4 cases (4/11, 36.36%). IgA decrease, IgM decrease, IgE decrease, IgM increase and IgE increase were observed in 6 cases, 3 cases, 5 cases, 3 cases, and 2 cases, respectively.Both T-lymphocyte and B-lymphocyte counts decreases was observed in 1 case.Genetic testing was performed in all recruited children, and genetic variations were detected in all of them.Inborn errors of immunity (IEIs) were diagnosed in 8 cases, including 4 diagnosed as CD 40 ligand deficiency with CD40LG variation, 1 of severe combined immunodeficiency with IL2RG variation, 1 of Signal transduction and activator of transcription 3(STAT3)-hyper-IgE syndrome with STAT3 variation and 1 of familial candidiasis type 2 with CARD9 compound heterozygous mutations.In the other 3 cases, 2 carried genetic variations that were likely pathogenic, and 1 case was considered uncertain. Conclusions:The clinical manifestations of HIV-negative children with TM infection are atypical, which is characterized as serious complications and high mortality.Early identification and gene testing to detect potential IEIs can improve the prognosis of TM infection.

Objective:To analyze the risk factors for hospital acquired pulmonary infections in elderly lung cancer patients during chemotherapy, and construct a prediction model to provide personalized guidance for clinical nursing work.Methods:Using convenience sampling, clinical case data of 405 elderly lung cancer patients who underwent chemotherapy at the First Affiliated Hospital of Zhengzhou University from January 2021 to December 2022 were collected. A total of 226 patients from January to December 2021 were selected as the modeling group, and 179 patients from January to December 2022 were selected as the validation group. In the modeling group, the patients were divided into two groups based on whether or not they had hospital acquired pulmonary infection, including the hospital acquired pulmonary infection group and the non-hospital acquired pulmonary infection group. Single factor analysis and multivariate Logistic regression analysis were used to determine the risk factors for hospital acquired pulmonary infection in elderly lung cancer patients during chemotherapy. A risk prediction model for hospital acquired pulmonary infection in elderly lung cancer patients during chemotherapy was constructed based on the results of multivariate analysis. The area under the receiver operating characteristic (ROC) curve was used to evaluate the predictive performance of the prediction model, and a calibration map was drawn for consistency testing. The prediction model was validated in the validation group to evaluate its predictive performance.Results:The incidence of hospital acquired pulmonary infection in the modeling group was 14.6% (33/226). The results of multivariate Logistic regression analysis showed that basic diseases, tumor-node-metastasis (TNM) staging, neutropenia, and invasive procedures were risk factors for hospital acquired pulmonary infections in elderly lung cancer patients undergoing chemotherapy ( P<0.05). Based on this result, a prediction model was constructed, with an area under the ROC curve of 0.832 [95% CI (0.764, 0.900) ] in the modeling group and 0.759 [95% CI (0.695, 0.824) ] in the validation group. The calibration curves of the modeling and validation groups showed a good coincidence between the predicted results and the actual results. Conclusions:Basic diseases, neutropenia, invasive procedures, and advanced stages of tumors are risk factors for hospital acquired pulmonary infections in elderly lung cancer patients undergoing chemotherapy. For high-risk patients, individualized intervention should be carried out as soon as possible to reduce the incidence of hospital acquired pulmonary infection.

Penicillium marneffei (PM) is the only thermally dimorphic species as a Penicillium that causes the transmitted penicilliosis marneffei (PSM). PM infection is more common in immunodeficient children.Due to the lack of specific clinical manifestations, PSM is easily misdiagnosed as pulmonary plasmacycosis, tuberculosis or pulmonary aspergillosis.Because of the rapid progress and multiple complications of transmitted PSM, its mortality remains high without timely and effective treatment.Therefore, early diagnosis and treatment are of great significance, and the diagnosis of PSM mainly relies on the immune function examination, measurement of serum markers, pathogen detection and imaging examinations.There is no standard antifungal regimen, and Amphoterus B and Itraconazole are used as the first-line treatment.This study aims to review the progress of diagnosis and treatment of PM infection in children.

Objective:To observe the therapeutic timing and dosage of intravenous immunoglobulin (IVIG) in children with severe adenovirus pneumonia.Methods:Clinical data of children with severe adenovirus pneumonia treated with IVIG at the Department of Respiratory, Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from January 2019 to January 2020 were retrospectively analyzed.Participants were classified as early presenters (5-10 days of illness course) and later presenters (11-15 days of illness course) according to the timing of IVIG treatment.They were further subdivided into plan 1 group[1 g/(kg·d) IVIG for 2 days] and plan 2 group [0.4-0.5 g/(kg·d) IVIG for 3-5 days]. Continuous variables and categorical variables between groups were analyzed by the nonparametric Mann- Whitney U test and the Fisher′ s exact test, respectively. Results:A total of 202 patients with the median age of 12 (12, 36) months were recruited, involving 128 early presenters (63.37%) and 74 later presen-ters (36.63%). Later presenters had a longer duration of fever [18.00(14.00, 23.25) days vs.11.00(9.00, 14.00) days], more demands for mechanical ventilation (33.78% vs.20.31%), and higher incidence of bronchiectasis (9.46% vs.1.56%) than those of early presenters (all P<0.05). For early presenters, no significant differences were detected in the demand for advanced life support, outcomes and sequelae between plan 1 group and plan 2 group (all P>0.05). For later presenters, a shorter duration of fever [18.00(14.00, 21.00) days vs.21.00(15.50, 30.75) days] and lower usage of extracorporeal membrane oxygenation (ECMO, 2.13% vs.18.52%) were observed in the plan 1 group than that of plan 2 group (all P<0.05). The incidence of post-infectious bronchiolitis obliterans and bronchiectasis as pulmonary sequelae was comparable between plan 1 group and plan 2 group ( P>0.05). The incidence of adverse events was 5.77% during IVIG infusion, showing no significant difference between plan 1 group and plan 2 group ( P>0.05). Conclusions:Early treatment of IVIG are very important to improve the prognosis of children with severe adenovirus pneumonia.For later presenters, a high dosage of IVIG is effective in reducing the ECMO use and shortening the duration of fever, thus providing clinical benefits.

Objective:To explore the technical feasibility of CT feature tracking (CT-FT) technique in evaluating left ventricular myocardial strain and evaluate the change of myocardial strain in patients with coronary heart disease.Methods:Eighty-one patients with coronary heart disease (lesion group) and 33 patients with normal coronary artery (control group) matched with age and sex were collected retrospectively from the Second Affiliated Hospital of Nanchang University from April 2019 to October 2020. The lesion group was first divided into single vessel stenosis group (42 cases) and multi vessel stenosis group (39 cases) according to the number of coronary artery stenosis branches, and the global myocardial strains of the left ventricle between the groups were analyzed. Lesion site included the left anterior descending branch (LAD), right coronary artery (RCA) and left circumflex branch (LCX), respectively. According to the degree of vascular stenosis, the lesion groups were divided into normal group, mild stenosis group, moderate stenosis group and severe stenosis group. The segmental myocardial strains of the branch segment of LAD, RCA or LCX were analyzed between groups. All CCTA examinations were performed with retrospective electrocardiogram gating. CVI 42 cardiac postprocessing software was used to obtain myocardial strain parameters, including global peak longitudinal strain (GPLS), global peak circumferential strain (GPCS), global peak radial strain (GPRS), and the segmental myocardial strains of the branch segment of LAD, RCA or LCX. The segmental myocardial strains included the peak longitudinal strain (PLS), peak circumferential strain (PCS) and peak radial strain (PRS). One way ANOVA or Kruskal Wallis H test were used for multi group analysis. Results:With the increased number of coronary artery stenosis branches, the absolute value of GPLS gradually decreased. The GPLS of the control group, single vessel stenosis group and multi vessel stenosis group were -14.1%±2.7%, -11.5%±2.3% and -8.8%±2.0%, respectively. The difference of GPLS between the 3 groups or any 2 groups was statistically significant (all P<0.001). The absolute values of GPRS and GPRS in multi vessel stenosis group were significantly lower than those in control group and single vessel stenosis group (all P<0.001). There was no significant difference in GPRS or GPRS between single vessel stenosis group and control group ( P=0.083, 0.118). And there were significant differences in the segmental myocardial strains of the branch segment of LAD, RCA or LCX among 3 groups ( P<0.001). In severe stenosis group, the absolute values of PRS, PCS and PLS in LAD, RCA or LCX were significantly lower than those in moderate stenosis group, mild stenosis group and normal group (all P<0.05). In the moderate stenosis group, the absolute value of PLS in each branch segment was lower than that of the mild stenosis and normal group (all P<0.05), and there was no significant difference in any 2 other myocardial strain parameters of each branch (all P>0.05). Conclusions:CT-FT technique was feasible to evaluate left ventricular myocardial function. With the increased number or degree of coronary artery stenosis, the global and segmental myocardial strain parameters of left ventricle gradually decreased, and the longitudinal strain was more sensitive.

Objective:To evaluate the efficacy and safety of nimotuzumab combined with definitive radiotherapy in the treatment of inoperable locally advanced oral and maxillofacial squamous cell carcinoma.Methods:A total of 33 patients with inoperable locally advanced oral and maxillofacial squamous cell carcinoma admitted to the Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from March 2015 to December 2019 were retrospectively selected as the research objects. The treatment regimen was all targeted therapy (nimotuzumab) combined with definitive radiotherapy, with or without chemotherapy, and the efficacy and safety of the treatment were analyzed. The primary endpoints were optimal response and overall survival (OS) , and the secondary endpoints were optimal duration of response (DOR) and progression-free survival (PFS) . The survival curve was drawn using the Kaplan-Meier method, the survival rate of the patients was analyzed, and the related adverse reactions were counted.Results:Of the 33 patients, there were 20 cases of complete remission (CR) , 5 cases of partial remission (PR) , 5 cases of stable disease (SD) , 2 cases of progressive disease (PD) , and 1 case could not be evaluated. The objective response rate was 75.8% (25/33) , and the disease control rate was 90.9% (30/33) . The mean OS of all cases was 54.5 months, and the 5-year OS rate was 57.0%. The mean DOR of the overall cases was 57.2 months, and the 5-year DOR rate was 64.4%. The mean PFS of the overall cases was 54.4 months, and the 5-year PFS rate was 59.8%. The 5-year OS rates of CR, PR and SD patients were 83.6%, 20.0% and 0 ( χ2=20.07, P<0.001) , the 5-year DOR rates were 85.0%, 20.0% and 0 ( χ2=16.89, P<0.001) , and the 5-year PFS rates were 84.0%, 20.0% and 0 ( χ2=15.91, P<0.001) . The OS, DOR and PFS of patients with CR were significantly better than those of patients with PR and SD (all P<0.05) . The 5-year OS rates of patients with oropharyngeal cancer and oral cancer were 62.5% and 40.6% ( χ2=1.67, P=0.197) , the 5-year DOR rates were 73.3% and 44.0% ( χ2=1.34, P=0.247) , and the 5-year PFS rates were 68.8% and 40.9% ( χ2=1.13, P=0.289) , with no statistically significant differences, but oropharyngeal cancer patients still showed a certain advantage. Common adverse reactions included oral mucositis and hematological toxicity, most of which were grade 1-2. Two (6.1%) patients had rash, and two (6.1%) patients had nausea and vomiting, which were considered to be related to nimotuzumab. All adverse reactions were relieved after symptomatic treatments. Conclusion:For patients with locally advanced oral and maxillofacial squamous cell carcinoma who are not suitable for surgery, the choice of nimotuzumab combined with definitive radiotherapy has a relatively satisfactory efficacy and survival rate, with good safety and high clinical value.