Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

Objective To investigate the antitumor effect of genetically engineered bacterial vesicles loaded with the photosensitizer Ce6.Methods Genetic engineering techniques were used to overexpress interferon α(IFNα)in Escherichia coli Nissle 1917(EcN),and the corresponding bacterial vesicles(BVs-IFNα)were extracted.The photosensitizer Ce6 was loaded onto the surface of the bacterial vesicles via ultrasonication to obtain BVs-IFNα@Ce6 nanoparticles.The BVs-IFNα@Ce6 complex was characterized using dynamic light scattering,nanoparticle tracking technology,confocal imaging,and ultraviolet absorption spectroscopy.The obtained BVs-IFNα@Ce6 nanoparticles were co-cultured with mouse melanoma cells(B16F10)and mouse bone marrow-derived dendritic cells(BMDC).The efficacy of photodynamic therapy was assessed using the CCK-8 method,and its ability to induce immune activation was verified using flow cytometry.Results The BVs-IFNα@Ce6 nanoparticles were successfully prepared with a stable particle size of around 180 nm.Under red light irradiation,BVs-IFNα@Ce6 exhibited significant cytotoxicity against B16F10 tumor cells and effectively promoted the expressions of immune-related molecules MHC-II,CD80/CD86 on the surface of BMDC cells,demonstrating its remarkable ability to induce immune activation.Conclusion The BVs-IFNα@Ce6 nanoparticles possess dual functions of enhancing photodynamic therapy efficacy and activating antitumor immunity,showing potential for clinical translation.

Objective To investigate the antitumor effect of genetically engineered bacterial vesicles loaded with the photosensitizer Ce6.Methods Genetic engineering techniques were used to overexpress interferon α(IFNα)in Escherichia coli Nissle 1917(EcN),and the corresponding bacterial vesicles(BVs-IFNα)were extracted.The photosensitizer Ce6 was loaded onto the surface of the bacterial vesicles via ultrasonication to obtain BVs-IFNα@Ce6 nanoparticles.The BVs-IFNα@Ce6 complex was characterized using dynamic light scattering,nanoparticle tracking technology,confocal imaging,and ultraviolet absorption spectroscopy.The obtained BVs-IFNα@Ce6 nanoparticles were co-cultured with mouse melanoma cells(B16F10)and mouse bone marrow-derived dendritic cells(BMDC).The efficacy of photodynamic therapy was assessed using the CCK-8 method,and its ability to induce immune activation was verified using flow cytometry.Results The BVs-IFNα@Ce6 nanoparticles were successfully prepared with a stable particle size of around 180 nm.Under red light irradiation,BVs-IFNα@Ce6 exhibited significant cytotoxicity against B16F10 tumor cells and effectively promoted the expressions of immune-related molecules MHC-II,CD80/CD86 on the surface of BMDC cells,demonstrating its remarkable ability to induce immune activation.Conclusion The BVs-IFNα@Ce6 nanoparticles possess dual functions of enhancing photodynamic therapy efficacy and activating antitumor immunity,showing potential for clinical translation.

Research into human drug metabolism,liver disease modeling as well as drug screening heavily depend on cell cultures for in vitro validation and experimental animal models for in vivo analysis.However,these methods are not without limitations.Cell stability is often compromised in vitro,and the species-specific differences between experimental animals and humans restrict the accuracy of these models in replicating in vivo drug metabolism and the pathophysiological features of human liver diseases.Humanized liver chimeric mouse models,developed through the transplantation and expan-sion of human liver cells into the livers of live mice,replicate the specific functionalities of the human liver.This replication is instrumental in fulfilling a variety of research objectives,including the replication of hepatitis viruses within mice,the modeling of liver metabolic disorders,the screening of liver cell-targeted therapeutic agents,and the evaluation of drug-induced hepatotoxicity.As such,these models are regarded as optimal for the preclinical validation of drug efficacy and safety assessment.This paper presents a comprehensive review and a vision of three representative humanized liver chimeric mouse models in terms of classification,construction principles,limitations,and current applications,thereby offering reference for the selection of appropriate models in research.

Objective:To assess respondents' understanding of the items in the Neck Dissection Impairment Index (NDII) through cognitive interviews and to revise the items accordingly.Methods:Totally 30 postoperative neck dissection patients returning for follow-up at Department of Otolaryngology-Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University from November 2023 to February 2024, were selected by purposive sampling for two rounds of cognitive interviews. Based on the interview results, the Chinese version of the NDII was revised.Results:The Chinese version of the NDII included ten items. The first round of interviews identified issues such as "incomplete expression, abstract language, and written form, " leading to revisions of five items. The second round of interviews showed that all respondents understood the revised items, and no new suggestions for modification were made.Conclusions:Cognitive interviews can improve respondents' comprehension of the Chinese version of the NDII, identify cognitive biases caused by cultural differences and improper linguistic expressions during the translation process, and enhance the understanding and acceptance of the scale's content among the target population.

For a long time, macrolides have been the first choice for the antibacterial treatment for pertussis.However, in the past decade, resistance to macrolide antimicrobials has been common in clinically isolated Bordetella pertussis in China, which is in contradiction with the recommended macrolide treatment.Therefore, Trimethoprim-Sulfamethoxazole (TMP-SMZ) is suggested as the first choice for antibacterial treatment for pertussis in China, with a dosage determined according to age and body weight, lasting 14 days.If TMP-SMZ cannot be used, full-dose and full-course β-lactam antimicrobials may be used, of which the effects should be assessed carefully.The impact of other antibacterial drugs, such as quinolones and tetracyclines, on the elimination of Bordetella pertussis should also be evaluated as soon as possible to treat adult pertussis and potential cases caused by drug-resistant bacteria in future.

Objective:To determine the erythromycin resistance of Bordetella pertussis isolates and their ptxP1 and ptxP3 phenotypic composition and compare clinical manifestations of children with pertussis caused by the two types of strains. Methods:This was a cross-sectional study, the pertussis cases diagnosed using bacterial culture from January 2019 to December 2022 in Beijing Children′s Hospital and the First People′s Hospital of Wuhu were collected.Any suspected Bordetella pertussis colonies were identified by the slide agglutination test.The susceptibility of isolates to erythromycin was detected by the E-test and K-B test.The ptxP gene was amplified by polymerase chain reaction and sequenced to determine its genotype. t-test, Mann-Whitney U-test, Chi-square test and Fisher′s exact test were use to statistical analysis. Results:A total of 192 strains of Bordetella pertussis were identified, including 188 (97.9%) erythromycin-resistant strains.Among the 188 strains, 30.3%(57/188) belonged to the ptxP1 genotype and 69.7%(131/188) belonged to the ptxP3 genotype.In children aged below 1 year old, the incidence of paroxysmal cough caused by infection with the ptxP3 strain was higher than that with the ptxP1 strain (57.1% vs.29.4%, P<0.05), and children infected with the ptxP3 strain were more likely to develop apnea or asphyxia (23.8% vs.17.6%), post-tussive vomiting (44.4% vs.32.4%), whooping cough (72.0% vs.50.0%) and pneumonia or bronchitis (85.7% vs.73.5%) compared to those infected with the ptxP1 strain, but the differences were not statistically significant(all P>0.05). In children aged 1 year old and above, the white blood cell count of children infected with the ptxP1 strain was higher than that of infections with the ptxP3 strain [13.5(9.9, 24.5)×10 9/L, 10.3 (7.0, 16.4)×10 9/L, P<0.05], and children infected with the ptxP1 strain were more likely to contract other pathogen infections than those infected with the ptxP3 strain (17.4% vs.4.4%, P>0.05). Conclusions:ptxP3 erythromycin-resistant Bordetella pertussis has become the main pathogen of pertussis.Infants with pertussis caused by the ptxP3 erythromycin-resistant strain show more significant manifestations and a higher possibility of severe symptoms than those infected with the ptxP1 erythromycin-resistant strain.

Objective:Preliminary explore the safety and efficacy of using only vitamin K antagonists without antiplatelet therapy after left ventricular assist devices (LVAD) implantation.Methods:This is a cohort study. Patients who underwent HeartCon LVAD implantation in TEDA International Cardiovascular Hospital from September 2020 to September 2022 were included. Oral warfarin sodium was given on postoperative days 1 to 2, with the target international standardized ratio (INR) of 2.0 to 2.5. Follow-up until September 2022, survival, INR level and occurrence of bleeding and thrombosis were recorded. Survival analysis was performed by the Kaplan-Meier method (censored for heart transplantation).Results:A total of 22 patients, including 16 male patients (72.7%), aged (51.0±13.3) years, were included. The duration of HeartCon LVAD support was (458±166) days and the INR during support was 2.28±0.26. One patient underwent the heart transplant at 307 d after implantation. One patient (4.5%) occured cardiac tamponade, two patients (9.1%) occured hemorrhagic stroke, five patients (22.7%) occured gastrointestinal bleeding, four patients (18.2%) occured gingival hemorrhage, two patients (9.1%) occured epistaxis, one patient (4.5%) occurred ischemic stroke, one patient (4.5%) occured pump thrombosis, and one patient (4.5%) occured aortic valve thrombosis. The survival rates were 100%, 95%, 95%, and 95% at 3 months, 6 months, 1 year, 2 years after implantation respectively.Conclusion:The single antithrombotic strategy using warfarin (target INR 2.0-2.5) without antiplatelet for patients with implantations of HeartCon type LVAD may be safe and effective.

The long-acting cabotegravir and rilpivirine injection regimen（CAB+RPV regimen）is the first approved long-acting antiretroviral therapy（ART）for HIV in China，administered once every two months. This regimen provides an innovative alternative to daily oral ART，benefiting virologically suppressed patients. Several large clinical-studies have shown that the CAB+RPV regimen achieves comparable virologic suppression and safety to daily oral regimens，while significantly enhancing patient satisfaction. Based on international and domestic HIV/AIDs guidelines and clinical evidence，this consensus offers expert recommendations on patient selection，clinical management，and key communication strategies for healthcare providers to support the effective use of this regimen，aiming to improve quality of life for people living with HIV and accumulate domestic clinical experience with this advanced treatment approach.

The long-acting cabotegravir and rilpivirine injection regimen（CAB+RPV regimen）is the first approved long-acting antiretroviral therapy（ART）for HIV in China，administered once every two months. This regimen provides an innovative alternative to daily oral ART，benefiting virologically suppressed patients. Several large clinical-studies have shown that the CAB+RPV regimen achieves comparable virologic suppression and safety to daily oral regimens，while significantly enhancing patient satisfaction. Based on international and domestic HIV/AIDs guidelines and clinical evidence，this consensus offers expert recommendations on patient selection，clinical management，and key communication strategies for healthcare providers to support the effective use of this regimen，aiming to improve quality of life for people living with HIV and accumulate domestic clinical experience with this advanced treatment approach.