Objective To screen the anti-CD22-specific nanobodies to provide a basis for immunotherapy agents. Methods The naive phage nanobody library was constructed and its diversity was analyzed. Three rounds of biotinylated streptavidin liquid phase screening were performed by using biotinylated CD22 antigen as the target, and the sequence of nanobodies against CD22 were identified by ELISA and gene sequencing. Results The capacity of the constructed naive phage nanobody library was 3.89×10⁹ CFU/mL, and the insertion of effective fragments was higher than 85%. Based on this library, seven anti-human CD22 nanobodies were screened, and the amino acid sequence comparison results showed that the overall similarity was 70.34%, and all of them were hydrophilic proteins. The results of protein-protein complex docking prediction showed that the mimetic proteins of the five nanobody sequences could be paired and linked to CD22, and the main forces were hydrophobic interaction and hydrogen bonding. Conclusion This study provided a basis for the study of chimeric antigen receptor T cells targeting CD22, successfully constructed the natural phage nanobody library and obtaining five anti-CD22-specific nanobodies.
Camelus/immunology* ; Single-Domain Antibodies/chemistry* ; Peptide Library ; Humans ; Animals ; Sialic Acid Binding Ig-like Lectin 2/genetics* ; Amino Acid Sequence ; Molecular Docking Simulation

BACKGROUND: Changes in N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels may not fully translate into patient-reported health status in patients with heart failure (HF). We aimed to evaluate the correlation between NT-proBNP levels and patient-reported health status changes at one month after discharge of patients, and their associations with risk of death and rehospitalization in patients with acute HF. METHODS: We used data from the China Patient-centered Evaluative Assessment of Cardiac Events Prospective Heart Failure Study (PEACE 5p-HF Study). Patient-reported health status was measured by the 12-item Kansas City Cardiomyopathy Questionnaire (KCCQ-12). Patients who were hospitalized for HF and completed the KCCQ-12 and NT-proBNP tests before and one month after discharge were eligible in our study. We stratified patients into different groups based on NT-proBNP levels (i.e., improved, stable, and deteriorated) and KCCQ-12 scores (i.e., not deteriorated and deteriorated). We also examined the associations of the joint NT-proBNP and KCCQ-12 change with the risk of one-year and four-year clinical outcomes. RESULTS: A total of 2461 patients were included in the analysis. The mean age was 64.06 ± 13.51 years, and 36.37% (895/2461) of the study population were female. Among patients with improved NT-proBNP levels, 115 (10.95%) patients had deteriorated KCCQ-12 scores. The correlation between the change in the KCCQ-12 score and NT-proBNP level was weak ( r2 = 0.002, P = 0.013). Stratification by changes in the KCCQ-12 score revealed subgroups with distinctive risks, such that patients with deteriorated KCCQ-12 scores in any of the NT-proBNP change groups exhibited an increased risk of one-year all-cause death than participants with not deteriorated KCCQ-12 scores in any of the NT-proBNP change groups. Patients with improved NT-proBNP levels and deteriorated KCCQ-12 scores presented greater risks of one-year all-cause death (hazard ratio [HR]: 2.45, 95% confidence interval [CI]: 1.34-4.48) than patients with stable NT-proBNP levels and not deteriorated KCCQ-12 scores (HR [95% CI], 1.77 [1.25-2.53]). CONCLUSIONS: A discrepancy between changes in NT-proBNP levels and KCCQ-12 scores was common. The change in NT-proBNP levels was not sufficient to characterize critical aspects related to HF during one month after discharge of patients. Changes in the KCCQ-12 score exhibit complementary information to NT-proBNP levels for the prediction of clinical outcomes in patients with acute HF. REGISTRATION www.clinicaltrials.gov (No. NCT02878811).
Aged ; Female ; Humans ; Male ; Middle Aged ; Health Status ; Heart Failure/metabolism* ; Natriuretic Peptide, Brain/metabolism* ; Peptide Fragments/metabolism* ; Prospective Studies

Objective:To investigate the impacts of total salvianolic acid(TSA)on macrophage polarization and mesenchymal macrophage infiltration in liver cancer bearing mice by regulating the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)pathway.Methods:Mice were randomly separated into liver cancer group,TSA low concentra-tion group,TSA medium concentration group,TSA high concentration group,TSA high concentration+TLR4 inhibitor(TAK242)group and control group,with 12 mice in each group.Except for control group,mice in all other groups were injected subcutaneously into the right axilla to construct liver cancer models by intraperitoneal transmission of third-generation H22 cell suspension.After suc-cessful modeling,administration was carried out once a day for 2 weeks.Changes in tumor weight,tumor volume,spleen index and thymus index were detected.Immunofluorescence was applied to detect proportions of CD86 and CD206 positive cells in tumor tissue.ELISA was applied to detect levels of inducible nitric oxide synthase(iNOS),IL-6 and IL-10 in tumor tissues.Flow cytometry was ap-plied to detect proportion of CD11b+F4/80+in tumor tissue.Western blot was applied to detect TLR4,p-NF-κB P65 and MyD88 pro-teins in tumor tissue.The life quality and survival rate of mice were observed.Results:Compared with control group,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were reduced in liver cancer group,while proportion of CD206 positive cells and level of IL-10 in tumor tissue were increased(P<0.05).Compared with liver cancer group,tumor weight and volume of mice in TSA low,medium,high concentration groups were decreased,while survival rate,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were increased,proportion of CD206 positive cells,level of IL-10,and proportion of CD11b+F4/80+in tumor tissue decreased(P<0.05).TAK242 reversed the effects of high concentration TSA on macrophage polarization and myeloid macrophage infiltration in liver cancer bearing mice.Conclusion:TSA may promote M1 polari-zation of macrophages in liver cancer bearing mice and inhibit myeloid macrophage infiltration by activating the TLR4/MyD88/NF-κB pathway.

Objective:To investigate the impacts of total salvianolic acid(TSA)on macrophage polarization and mesenchymal macrophage infiltration in liver cancer bearing mice by regulating the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)pathway.Methods:Mice were randomly separated into liver cancer group,TSA low concentra-tion group,TSA medium concentration group,TSA high concentration group,TSA high concentration+TLR4 inhibitor(TAK242)group and control group,with 12 mice in each group.Except for control group,mice in all other groups were injected subcutaneously into the right axilla to construct liver cancer models by intraperitoneal transmission of third-generation H22 cell suspension.After suc-cessful modeling,administration was carried out once a day for 2 weeks.Changes in tumor weight,tumor volume,spleen index and thymus index were detected.Immunofluorescence was applied to detect proportions of CD86 and CD206 positive cells in tumor tissue.ELISA was applied to detect levels of inducible nitric oxide synthase(iNOS),IL-6 and IL-10 in tumor tissues.Flow cytometry was ap-plied to detect proportion of CD11b+F4/80+in tumor tissue.Western blot was applied to detect TLR4,p-NF-κB P65 and MyD88 pro-teins in tumor tissue.The life quality and survival rate of mice were observed.Results:Compared with control group,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were reduced in liver cancer group,while proportion of CD206 positive cells and level of IL-10 in tumor tissue were increased(P<0.05).Compared with liver cancer group,tumor weight and volume of mice in TSA low,medium,high concentration groups were decreased,while survival rate,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were increased,proportion of CD206 positive cells,level of IL-10,and proportion of CD11b+F4/80+in tumor tissue decreased(P<0.05).TAK242 reversed the effects of high concentration TSA on macrophage polarization and myeloid macrophage infiltration in liver cancer bearing mice.Conclusion:TSA may promote M1 polari-zation of macrophages in liver cancer bearing mice and inhibit myeloid macrophage infiltration by activating the TLR4/MyD88/NF-κB pathway.

Humans ; Aged ; Lung Diseases ; Pneumonia/drug therapy* ; Adrenal Cortex Hormones/therapeutic use* ; Pulmonary Disease, Chronic Obstructive/drug therapy* ; Administration, Inhalation ; Community-Acquired Infections/drug therapy*

AIM: To analyze the causes of blindness and low vision in patients with visual disability in Yangpu District of Shanghai from 2019 to 2022.METHODS:Cross-sectional study. A total of 1 604 patients who participated in the evaluation of visual disability in Shanghai Yangpu District Kongjiang Hospital, from April 2019 to December 2022 were selected for the study. The grade of visual disability and the main causes of blindness and low vision were determined by trained doctors.RESULTS:A total of 804 patients with visual disabilities were identified, with 87.31% aged 60 and above. The causes of visual disability were high myopic retinopathy(30.47%), age-related macular degeneration(23.26%), glaucoma(17.04%), and diabetic retinopathy(11.07%). Glaucoma(36.96%)is the leading cause of blindness.CONCLUSION: The majority of patients with visual disability are aged 60 years and above. More attention should be paid to the elderly population. Comprehensive prevention, treatment and rehabilitation measures should be applied in different diseases based on classification, so as to early reduce the occurrence of visual disability.

Objective:To investigate the effect of water intake on bladder filling time before embryo transfer.Methods:A total of 189 patients were collected from February to June 2023 who were to undergo embryo transfer in Guangdong Women and Children Hospital. The patients were divided into group A ( n=61), group B ( n=64) and group C ( n=64) using a random number table and they were respectively given 300 mL, 500 mL and 700 mL water to drink. Abdominal ultrasound was performed every 15 min, a total of 1-5 times, from 45 min after drinking water until the bladder filling. The bladder filling time and bladder volume were collected. Kaplan-Meier method was used to compare the difference of bladder filling time among the three groups. The multivariate Cox regression was used to analyze factors of bladder filling time. Results:The cumulative bladder filling rates of group A, group B and group C at 105 min after drinking water were 57.4% (35/61), 90.6% (58/64) and 98.4% (63/64), respectively, and the median survival time (95% CI) of bladder filling was 105.0 (89.9-120.1) min, 60.0 (55.4-64.7) min and 60.0 (55.4-64.6) min, respectively. Pairwise comparison of Kaplan-Meier analysis revealed that the bladder filling time of group A was longer than that of group B and group C ( P<0.001), and there was no statistically significant difference between group B and group C ( P>0.05). The results of age-stratification analysis showed that the bladder filling time of younger patients in group A [90.0 (75.2-104.8) min] was longer than that in group B [60.0 (55.8-64.2) min, P<0.001] and group C [60.0 (55.1-64.8) min, P<0.001], and there was no statistical significance between group B and group C ( P>0.05); the bladder filling time of older patients in group C [60.0 (59.1-70.9) min] was shorter than that in group A [105.0 (89.9-120.1) min, P<0.001] and group B [75.0 (64.3-85.7) min, P=0.027], there was no statistical significance between group A and group B ( P>0.05). Multivariate Cox regression analysis showed that taking group A as reference, the hazard ratio ( HR, 95% CI) of groups B and C were 2.71 (1.78-4.21) and 3.23 (2.10-4.96), both P<0.001. The HR (95% CI) of the elderly patients was 0.69 (0.49-0.99), P=0.044. Conclusion:Water intake and age are independent factors affecting bladder filling time in embryo transfer patients. Patients are recommended to drink 500 mL of water 75 min before embryo transfer and appropriately increase the amount of water or extend the bladder preparation time after drinking water for elderly patients.

Objective:To investigate the effect of water intake on bladder filling time before embryo transfer.Methods:A total of 189 patients were collected from February to June 2023 who were to undergo embryo transfer in Guangdong Women and Children Hospital. The patients were divided into group A ( n=61), group B ( n=64) and group C ( n=64) using a random number table and they were respectively given 300 mL, 500 mL and 700 mL water to drink. Abdominal ultrasound was performed every 15 min, a total of 1-5 times, from 45 min after drinking water until the bladder filling. The bladder filling time and bladder volume were collected. Kaplan-Meier method was used to compare the difference of bladder filling time among the three groups. The multivariate Cox regression was used to analyze factors of bladder filling time. Results:The cumulative bladder filling rates of group A, group B and group C at 105 min after drinking water were 57.4% (35/61), 90.6% (58/64) and 98.4% (63/64), respectively, and the median survival time (95% CI) of bladder filling was 105.0 (89.9-120.1) min, 60.0 (55.4-64.7) min and 60.0 (55.4-64.6) min, respectively. Pairwise comparison of Kaplan-Meier analysis revealed that the bladder filling time of group A was longer than that of group B and group C ( P<0.001), and there was no statistically significant difference between group B and group C ( P>0.05). The results of age-stratification analysis showed that the bladder filling time of younger patients in group A [90.0 (75.2-104.8) min] was longer than that in group B [60.0 (55.8-64.2) min, P<0.001] and group C [60.0 (55.1-64.8) min, P<0.001], and there was no statistical significance between group B and group C ( P>0.05); the bladder filling time of older patients in group C [60.0 (59.1-70.9) min] was shorter than that in group A [105.0 (89.9-120.1) min, P<0.001] and group B [75.0 (64.3-85.7) min, P=0.027], there was no statistical significance between group A and group B ( P>0.05). Multivariate Cox regression analysis showed that taking group A as reference, the hazard ratio ( HR, 95% CI) of groups B and C were 2.71 (1.78-4.21) and 3.23 (2.10-4.96), both P<0.001. The HR (95% CI) of the elderly patients was 0.69 (0.49-0.99), P=0.044. Conclusion:Water intake and age are independent factors affecting bladder filling time in embryo transfer patients. Patients are recommended to drink 500 mL of water 75 min before embryo transfer and appropriately increase the amount of water or extend the bladder preparation time after drinking water for elderly patients.

Objective:To explore the selection strategy of blastocyst transfer number in freeze-thaw cycle for women over 40 years old, so as to provide reference for reducing twin rate and improving perinatal clinical outcome.Methods:A retrospective analysis was made of 377 patients who underwent freeze-thaw blastocyst transplantation in the reproductive center of Guangdong Maternal and Child Health Hospital from January 2017 to December 2019. They were divided into single blastocyst and double blastocyst transplantatio groups according to the number of blastocyst transplantation. The clinical pregnancy rate, implantation rate, abortion rate, live birth rate, premature delivery rate, twin rate and singleton delivery rate were compared between the two groups.Results:⑴There was no significant difference between two groups regarding the majority of baseline characteristics, including age at retrieval, age at transfer, body mass index (BMI), antral follicle count (AFC), basal follicle stimulating hormone (FSH), anti Mullerian hormone (AMH), endometrial thickness at transfer day, number of oocytes retrieved, Gn starting dose, Gn days, Gn dosage, embryos at cleavage stage and top-quality embryos ( P>0.05). ⑵ There was no significant difference in the rate of implantation, early pregnancy loss, late pregnancy loss and live birth between two groups ( P>0.05). ⑶ The preterm birth rate was higher in the double blastocyst transplantation group compared with the single blastocyst transplantation group, albeit not reaching significant difference (31.7% vs 12.5%, P=0.083). ⑷ The clinical pregnancy rate and the twin pregnancy rate was significantly higher in the double blastocyst transplantation group compared with the single blastocyst transplantation group ( P<0.05). ⑸ The singleton birth rate was significantly lower in the double blastocyst transplantation group compared with the single blastocyst transplantation group (75.61% vs 95.83%, P<0.05). Conclusions:In women ≥40 years old, transferring a single blastocyst can result in live birth rate that is similar as transferring two blastocysts while dramatically reducing the risk of twin pregnancy rate and increasing singleton birth rate.

Objective:To evaluate the impact of short-term low-medium dose of corticosteroids on the clinical outcomes of patients with community-acquired pneumonia due to influenza A (FluA-CAP).Methods:This was a multicenter, retrospective study, including 693 patients hospitalized with FluA-CAP from Beijing Jishuitan Hospital, Qingdao Municipal Hospital, Beijing Huimin Hospital, Beijing Chao-Yang Hospital and the 2nd People′s Hospital of Yunnan Province during January 1, 2013 to December 31, 2018. The clinical characteristics of patients with or without corticosteroids administration were compared. The first dose of corticosteroids was administrated within 72 hours after admission, with the average dose of methylprednisolone (0.6±0.3) mg/(kg·d) and duration of (4.0±1.2) days. An adjusted logistic regression model was performed to assess the impact of corticosteroids treatment on the clinical outcomes (noninvasive ventilation, invasive ventilation, vasopressor use, admittance to intensive care unit (ICU), 30-day mortality, hyperglycemia needing insulin treatment and gastrointestinal bleeding). Mann-Whitney test and χ2 test were used for the statistical analysis. Results:Among the 693 patients, 132 patients received corticosteroids. Logistic regression analysis revealed that asthma (odd ratios ( OR)=15.528, 95% confidence interval ( CI) 1.953-123.484, P=0.01), chronic obstructive pulmonary disease ( OR=21.904, 95% CI 4.548-105.504, P<0.01) and arterial partial pressure of oxygen (PaO 2)/fraction of inspired oxygen (FiO 2)<300 mmHg (1 mmHg=0.133 kPa, OR=2.701, 95% CI 1.513-4.822, P<0.01) were independent risk factors for corticosteroids use in the FluA-CAP patients. An adjusted logistic regression model showed that low-medium dose corticosteroids administration was associated with decreased risks for early (defined as zero to three days after the first dose of corticosteroids) noninvasive ventilation ( OR=0.342, 95% CI 0.156-0.750, P<0.01), and increased risk for late (defined as four to 14 days after the first dose of corticosteroids) vasopressor use ( OR=2.651, 95% CI 1.913-6.306, P<0.01), late hyperglycemia which needed insulin treatment ( OR=9.739, 95% CI 2.174-21.769, P=0.019), ICU admission ( OR=3.075, 95% CI 1.166-8.143, P<0.01) and the 30-day mortality ( OR=2.372, 95% CI 1.337-4.549, P<0.01). In patients with asthma or chronic obstructive pulmonary disease ( OR=2.343, 95% CI 1.145-4.073, P<0.01) and PaO 2/FiO 2<300 mmHg ( OR=1.961, 95% CI 1.029-4.212, P<0.01), corticosteroids administration increased the risk of 30-day mortality. Conclusion:Low-medium corticosteroids treatment is associated with poor outcomes of FluA-CAP patients, and is not recommended to be used routinely.