OBJECTIVE To investigate the impact of vesicular stomatitis virus envelope glycopro-tein-G(VSV-G)modification on the mucosal immune efficacy of antigen-loaded engineered exosome vaccines.METHODS In vitro experiments:Dendritic cells(DCs)were divided into three groups:cell-control(treated with culture medium),receptor binding domain(RBD)(transfected with plasmid RBD),and RBD+VSV-G(co-transfected with plasmids RBD and VSV-G).Expression levels of RBD and VSV-G were assessed using Western blotting,flow cytometry,and immunofluorescence.Exosomes were extracted via ultracentrifugation,whose morphology,size distribution,and marker proteins were analyzed using transmission electron microscopy,nanoparticle tracking analysis,and Western blotting that confirmed the expressions of RBD and VSV-G in the exosomes.In vivo experiments:① Female BALB/c mice were divided into the control group Mock exosomes(Mock-Exo)(derived from the supernatant of cell-control),RBD decorated exosomes(RBD-Exo)(derived from the RBD cell supernatant),and RBD and VSV-G decorated exosomes(RBD+VSV-G-Exo)(derived from RBD+VSV-G cell supernatant).Follow-ing intranasal immunization with the respective vaccines,the nasal retention effects were evaluated using in vivo imaging.Flow cytometry was used to assess the ability to recruit immune cells to the nasal tissue.Serum RBD-specific immunoglobulin G(IgG)and mucosal immunoglobulin A(IgA)(bronchoal-veolar lavage fluid/nasal wash)were quantified at 7 and 21 d post-immunization by enzyme-linked immuno-sorbent assay.Body weight changes were monitored and key serum biochemical parameters along with histopathological damage to major organs were analyzed following immunization.② Female BALB/c mice were divided into the Mock-Exo group(intranasally inoculated with Mock-Exo),RBD+VSV-G-Exo group(intranasally inoculated with RBD+VSV-G-Exo),and RBD+VSV-G-Exo(im)group(intramus-cularly injected with RBD+VSV-G-Exo).RESULTS In vitro experiments:RBD and VSV-G were successfully expressed in cells,with positive rates of RBD+and VSV-G+cells at 64.4%and 31.2%,respectively.The extracted exosomes exhibited regular morphology and qualified purity,with a particle size of approximately 138 nm and successfully loaded RBD and VSV-G proteins.In vivo experiments:Compared to Mock-Exo and RBD-Exo,RBD+VSV-G-Exo prolonged nasal retention time to 96 h and markedly increased the numbers of CD49B+natural killer cells,CD11c+dendritic cells,and F4/80+macrophages in nasal tissues.RBD+VSV-G-Exo induced robust RBD-specific immune responses,with serum IgG titers,BALF IgA titers,and nasal wash IgA titers reaching 1∶5 215,1∶2 560,1∶1 114,respec-tively.In contrast,no RBD-specific IgA antibody titers were detected in the BALF and nasal wash of mice treated with RBD+VSV-G-Exo(im).Mice showed stable body weight gain during 30 d post-immu-nization.Major serum biochemical indices were within normal reference ranges,and no obvious patho-logical changes were observed in major organs or olfactory bulbs 7 d after immunization.CONCLU-SION VSV-G modification extends the retention time of engineered exosome vaccines in nasal tissues,enhance their ability to recruit immune cells,and induce a high-level antigen-specific respiratory mucosal immune response.

OBJECTIVE To investigate the impact of vesicular stomatitis virus envelope glycopro-tein-G(VSV-G)modification on the mucosal immune efficacy of antigen-loaded engineered exosome vaccines.METHODS In vitro experiments:Dendritic cells(DCs)were divided into three groups:cell-control(treated with culture medium),receptor binding domain(RBD)(transfected with plasmid RBD),and RBD+VSV-G(co-transfected with plasmids RBD and VSV-G).Expression levels of RBD and VSV-G were assessed using Western blotting,flow cytometry,and immunofluorescence.Exosomes were extracted via ultracentrifugation,whose morphology,size distribution,and marker proteins were analyzed using transmission electron microscopy,nanoparticle tracking analysis,and Western blotting that confirmed the expressions of RBD and VSV-G in the exosomes.In vivo experiments:① Female BALB/c mice were divided into the control group Mock exosomes(Mock-Exo)(derived from the supernatant of cell-control),RBD decorated exosomes(RBD-Exo)(derived from the RBD cell supernatant),and RBD and VSV-G decorated exosomes(RBD+VSV-G-Exo)(derived from RBD+VSV-G cell supernatant).Follow-ing intranasal immunization with the respective vaccines,the nasal retention effects were evaluated using in vivo imaging.Flow cytometry was used to assess the ability to recruit immune cells to the nasal tissue.Serum RBD-specific immunoglobulin G(IgG)and mucosal immunoglobulin A(IgA)(bronchoal-veolar lavage fluid/nasal wash)were quantified at 7 and 21 d post-immunization by enzyme-linked immuno-sorbent assay.Body weight changes were monitored and key serum biochemical parameters along with histopathological damage to major organs were analyzed following immunization.② Female BALB/c mice were divided into the Mock-Exo group(intranasally inoculated with Mock-Exo),RBD+VSV-G-Exo group(intranasally inoculated with RBD+VSV-G-Exo),and RBD+VSV-G-Exo(im)group(intramus-cularly injected with RBD+VSV-G-Exo).RESULTS In vitro experiments:RBD and VSV-G were successfully expressed in cells,with positive rates of RBD+and VSV-G+cells at 64.4%and 31.2%,respectively.The extracted exosomes exhibited regular morphology and qualified purity,with a particle size of approximately 138 nm and successfully loaded RBD and VSV-G proteins.In vivo experiments:Compared to Mock-Exo and RBD-Exo,RBD+VSV-G-Exo prolonged nasal retention time to 96 h and markedly increased the numbers of CD49B+natural killer cells,CD11c+dendritic cells,and F4/80+macrophages in nasal tissues.RBD+VSV-G-Exo induced robust RBD-specific immune responses,with serum IgG titers,BALF IgA titers,and nasal wash IgA titers reaching 1∶5 215,1∶2 560,1∶1 114,respec-tively.In contrast,no RBD-specific IgA antibody titers were detected in the BALF and nasal wash of mice treated with RBD+VSV-G-Exo(im).Mice showed stable body weight gain during 30 d post-immu-nization.Major serum biochemical indices were within normal reference ranges,and no obvious patho-logical changes were observed in major organs or olfactory bulbs 7 d after immunization.CONCLU-SION VSV-G modification extends the retention time of engineered exosome vaccines in nasal tissues,enhance their ability to recruit immune cells,and induce a high-level antigen-specific respiratory mucosal immune response.

Objective:To compare the histopathological features and treatment efficacy of different methods for metachronous early gastric cancer (MEGC) in the remnant stomach.Methods:A total of 66 patients [38 endoscopic submucosal dissection (ESD) and 28 gastrectomy] with MEGC in the remnant stomach from January 2014 to December 2020 in Drum Tower Hospital were divided into the ESD group and the gastrectomy group. The baseline characteristics, histopathological features, treatment efficacy, and cost differences of the two groups were analyzed.Results:The MEGC in the remnant stomach mostly occurred in elderly male patients, with the mean age of 69.7±8.5 years. The mean interval of the occurrence of MEGC in the remnant stomach was 6 years. As for the tumor location, the gastric body (31.6%) was the main location in the ESD group and gastric cardia (53.6%) in the gastrectomy group with significant difference ( χ2=11.07, P=0.026). The mean operation time, hospital stay, postoperative fasting time, and total treatment cost were 80.0 min, 6.0 d, 1.5 d, ￥19 436 in the ESD group and 215.0 min, 19.0 d, 6.5 d, and ￥68 665 in the gastrectomy group, respectively, with significant differences between the two groups ( P<0.05). The overall survival rate during follow-up was 76.3% in the ESD group and 71.4% in the gastrectomy group with no significant difference between the two groups ( χ2=0.736, P=0.778). In terms of postoperative complications, the incidences of bleeding and infection were 7.9% and 5.3% in the ESD group, and those of obstruction and infection were both 14.3% in the gastrectomy group. There was significant difference in the incidences of postoperative obstruction between the two groups ( P<0.05). Conclusion:ESD is safe and effective for MEGC in the remnant stomach and is better than gastrectomy in terms of the treatment cost and operation time, but the long-term efficacy still needs to be validated by large-scale prospective studies.

Objective:To analyze the status of the dyadic coping in spouses of young and middle-age cervical cancer patients undergoing synchronous radiotherapy and chemotherapy, and to explore the influencing factors in bi-directional of patients and their spouses.Methods:With the convenience sampling method, a total of 150 cervical cancer patients undergoing synchronous radiotherapy and chemotherapy who were hospitalized in the Radiotherapy Department of Tianjin Medical University Cancer Institute and Hospital from March 2021 to February 2022 and their spouses were selected. A cross-sectional study was conducted by the Dyadic Coping Inventory, the Lock-Wallace Marital Adjustment Test, the General Self-Efficacy Scale, the Hospital Anxiety and Depression Scale, etc. Besides, multiple linear regression was used to identify predictors of the dyadic coping in spouses of cervical cancer patients undergoing synchronous radiotherapy and chemotherapy.Results:The total score of dyadic coping, marital quality, general self-efficacy, anxiety and depression in spouses of cervical cancer patients undergoing synchronous radiotherapy and chemotherapy were (121.69 ± 19.67), (97.23 ± 25.05), (25.13 ± 5.19), (9.98 ± 3.46), (8.19 ± 4.06) points. The scores of anxiety and depression of cervical cancer patients undergoing synchronous radiotherapy and chemotherapy were (10.57 ± 3.60), (9.10 ± 4.12) points. Multiple linear regression analysis showed that factors of the patients′ anxiety and depression, spouse′s perception of marital quality, spouse′s general self-efficacy, changes in family relationship, family income, and period of radiotherapy were the main influencing factors ( P<0.01), which accounted for 55.7% of total variation. Conclusions:The level of dyadic coping in spouses of cervical cancer patients undergoing synchronous radiotherapy and chemotherapy was medium, the marriage quality remains to be further improved,and its influence factors involved in the bi-directional of patient and spouse, including patients′ anxiety and depression, the changes of the family relationship, period of radiotherapy, spouses perception of marital quality and self-efficacy and family income. Clinical medical staff can improve the level of dyadic coping in spouses of cervical cancer patients undergoing synchronous radiotherapy and chemotherapy by improving their cognition of disease, reducing the incongruence of dyadic illness appraisals, and taking multiple measures to reduce the economic burden felt of the spouses.

Objective To study the current status of catheter - related thrombosis(CRT)in Chinese children through a retrospective analysis of the inpatients in the Department of Medicine,Beijing Children's Hospital Affiliated to Capital Medical University. Methods The clinical data of the inpatients with CRT from November 2010 to November 2013 were collected retrospectively,and the causes,clinical symptoms,diagnosis,treatment and prognosis were ana-lyzed. Results There were 42 cases of children with CRT in Beijing Children's Hospital Affiliated to Capital Medical University. Among the cases,the male to female ratio was 1. 0:0. 5;the median age of onset was 88(2 - 186)months with ﹤ 1 year old counted for 16. 7%(7 / 42 cases)and 13 - 14 years old counted for 11. 9%(5 / 42 cases);the distri-bution differences between the male and the female age were not significant(P = 0. 826). The median time from cathe-terization to CRT onset was 9(1 - 81)days,0 - 10 days after catheterization was the peak of onset(52. 5% ,21 / 40 ca-ses)followed by 10 - 20 days(35. 0% ,14 / 40 cases). The protopathy was usually hematologic tumor,kidney disease or deep fungal infection. Slightly more cases developed CRT on the right side(57. 1% ,24 / 42 cases)than on the left side (38. 1% ,18 / 42 cases). All cases were diagnosed by using B - ultrasound,of whom 28. 6%(12 / 42 cases)were symp-tom - free. After being diagnosed,7. 1%(3 / 42 cases)were treated with conservative methods such as immobilization of the affected limbs and hot compress;7. 1%(3 / 42 cases)had catheter removed;anticoagulant and/ or thrombolytics after catheter removal used in 33. 3% patients(14 / 42 cases). After 1 week,22 cases were reviewed,of whom 54. 5%(12 / 22 cases)had thrombosis reduced(all with intervention),thrombosis growing in 22. 7% patients(5 / 22 cases), and thrombosis did not change in 22. 7% patients(5 / 22 cases). Three cases needed re - catheterization after catheter removal,and all of 3 cases had CRT recurrences(100% ). Conclusions CRT is more common among infants and senior children. CRT usually develops within 20 days after catheterization. Children with hematologic tumor,kidney disease or deep fungal infection are more likely to have CRT. Routine ultrasound test should be conducted to monitor CRT in catheterized children. Once CRT is diagnosed,patients need to be treated with anticoagulants and/ or thrombo-lytics. Catheter should also be removed if necessary. Recatheterization can result in CRT recurrence.

Aim To study the mechanism of DXM and NAC inhibiting the expression of IL-8 and ICAM-1 in A549 cells. Methods The expression of IL-8 and ICAM-1 was detected by ELISA and flow cytometry re-spectively; the expression of GR,HDAC,AP-1,NF-κB was detected by Western blot, while the activity of HDAC was detected by spectrophotometry. ResultsThe increasing expression of IL-8 and ICAM-1 induced by TNF-α could be inhibited by DXM and NAC in A549 cells. DXM could inhibit the transcribed activa-tion of AP-1,NF-κB, and the expression of HDAC and its activity induced by TNF-α and LPS; NAC only in-hibited the transcribed activation of NF-κB, while it had no affection on the transcribed activation of AP-1 and the expression of HDAC and its activity. Conclu-sions DXM and NAC both have the anti-inflammatory effect. DXM plays the role of anti-inflammation through increasing the expression and activation of HDAC, in-hibiting the transcribed activation of AP-1 and NF-κB, while NAC has no effect on the expression and activa-tion of HDAC, which shows that NAC does not exert anti-inflammatory effect through acetylation signal.

Objective To investigate the effect of thrombospondin 1(TSP-1) on the cell proliferation of cultured rat pulmonary artery smooth cell (PASMCs) in vitro .Methods Rat PASMCs were cultured in vitro ,and treated with different concentrations (10-12 、10-11 、10-10 mol/L) of TSP-1 for 12 ,24 ,48 h .The cell proliferation was quantified by MTT assay .The cell cycle of PASMCs was measured by flow cytometric(FCM) analysis .Results MTT assay showed that TSP-1 promoted the proliferation of PASMCs significantly ,and the effect was concentration-dependent and time-dependent .FCM analysis indicated that TSP-1 increased the percentage of S phase .The percentage of S phase of PASMCs were increased after treated with thrombospondin-1 for 12 h , slight down after 24 h ,while reached a maximal level at 48 h .Conclusion The TSP-1 promotes rat PASMCs proliferation in a con-centration-dependent and time-dependent manner .

Objective To investigate current status of cognitive behavior on chronic obstructive pulmonary disease (COPD) to provide evidence for targeted intervention.Methods A questionnaire survey on COPD was conducted at 12 settings of 4 prefectures that were sampled from Ningxia Province using multistage stratified cluster random sampling method,and 4268 residents at least 40 years old were interviewed.Multivariate linear regression method was used for data analysis.Results A total of 4056 valid questionnaires were returned from 4200 participants with an effective response rate of 96.57％.Only 6.51％(264/4056) had an idea of COPD,13.88％ (563/4056) regarded cigarette smoking as a risk factor of chronic bronchitis and emphysema,and 6.39％ (259/4056) knew pulmonary function tests.Home income,living area,education level and ethnicity were main factors influencing COPD awareness.The percentage of current cigarette smoking was 28.80％ (1168/4056) with 20.40％ in Hui (Muslim) ethnic group and 34.63％ in Han ethnic group (P ＜ 0.05 ) ; smoking index and severe smokers were not significantly different between the two groups ( P ＞ 0.05 ).Conclusion Our data suggest a considerably poor knowledge and prevention awareness of COPD in Ningxia Province.In spite of relatively lower cigarette smoking rate,the problem that Hui (Muslim) ethnic people are lack of COPD awareness should not to be neglected.