Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To analyze the epidemiological characteristics and trend of hospitalization of the patients with herpes zoster in Beijing from 2017 to 2022.Methods:In this retrospective study, the information of hospitalization of herpes zoster patients were collected from all medical institutions at the first level and above in Xicheng, Changping, and Miyun districts of Beijing. The age and gender specific hospitalization rates and age-standardized hospitalization rates were calculated. Joinpoint regression model was used to explore the trend of the hospitalization rates, and the influencing factors of the hospital stay length and complications were analyzed.Results:The age-standardized hospitalization rate of the patients with herpes zoster was 10.82/100 000-18.43/100 000 in Beijing from 2017 to 2022 [annual percent change (APC) =5.86%, 95% CI: -2.80%-15.98%]. The age-standardized hospitalization rate of the cases with herpes zoster as the main diagnosis showed an upward trend (APC=11.35%, 95% CI: 7.21%-16.23%). The age-standardized hospitalization rate showed an upward trend in women (APC=14.34%, 95% CI: 7.95%-22.37%). The hospitalization rate showed a downward trend in age group 30-39 years (APC=-24.92%, 95% CI: -48.56% - -1.85%) and showed upward trends in age group 70-79 years and 80-109 years (APC=23.18%, 95% CI: 13.53%-35.58%; APC=4.90%, 95% CI: 1.18%-9.19%). Complications occurred in 66.28% (680/1 026) of the patients. The median hospital stay length was 9 (5,15) days, and the patients with high age (≥80 years) and two or more complications had longer hospital stay, which were 12 (6, 23) and 14 (7, 27) days respectively ( P<0.001). Conclusions:The hospitalization rate in women and the elderly aged ≥70 years with herpes zoster as the main diagnosis showed upward trends in Beijing in recent years. The elderly aged ≥80 years usually had longer hospital stay, showing a relatively disease burden level. More attention should be paid to development of intervention strategies, such as vaccine, for this population.

Objective To analyze changes in sleep,mood state,and brain function in healthy populations living in near-sea-level environments before and after exposure to high-altitude environment,and to explore the correlations between regional brain functional changes and variations in sleep and mood states.Methods A total of 45 healthy volunteers were enrolled.The participants came from regions of near-sea-level altitudes and were exposed to the high-altitude environment for a short period of time.The Pittsburgh Sleep Quality Index(PSQI),Zung Self-Rating Depression Scale(SDS),Patient Health Questionnaire-9(PHQ-9),Zung Self-Rating Anxiety Scale(SAS),and Generalized Anxiety Disorder-7(GAD-7)were administered to assess sleep quality as well as depressive and anxiety symptoms at 4 time points—prior to high-altitude exposure,immediately after exposure,one month after returning to low-altitude regions,and three months after returning to low-altitude regions.Resting-state functional magnetic resonance imaging(rs-fMRI)data were collected before and after high-altitude exposure,and regional brain functional parameters,including the amplitude of low-frequency fluctuations(ALFF)and functional connectivity strength,were analyzed.Statistical analyses were performed,including a linear mixed-effects model to evaluate longitudinal changes in scale scores,paired-sample t-tests to compare brain function differences before and after exposure,and Pearson correlation analyses to examine the relationship between brain functional changes and alterations in sleep and mood states.Results Compared with the pre-exposure findings,the participants exhibited significantly increased PSQI scores(8.89±4.41 vs.5.08±2.69,P<0.05)and PHQ-9 scores(3.60±4.19 vs.1.54±2.30,P<0.05)immediately after high-altitude exposure.One month after returning to the low-altitude environment,both sleep and depression scores decreased relative to the findings immediately after exposure(PSQI:3.88±2.13 vs.8.89±4.41,P<0.05;PHQ-9:1.50±2.25 vs.3.60±4.19,P<0.05)and showed no statistically significant difference compared with the pre-exposure findings(P>0.05).Three months after returning to near-sea-level environment,sleep,depression,and anxiety scores were all reduced compared with the findings immediately after exposure(PSQI:3.76±2.31 vs.8.89±4.41,P<0.05;PHQ-9:1.24±2.13 vs.3.60±4.19,P<0.05;SAS:23.84±5.93 vs.27.93±7.05,P<0.05),also showing no significant difference compared with the pre-exposure levels(P>0.05).Brain function analysis revealed that,relative to the pre-exposure levels,ALFF in the bilateral superior temporal gyrus,insula,and dorsolateral prefrontal cortex(DLPFC)increased after high-altitude exposure(P<0.05),and that functional connectivity strength in the DLPFC was also elevated(P<0.05).Furthermore,changes in DLPFC functional connectivity strength were positively correlated with changes in sleep and mood scores(P<0.05).Conclusion High-altitude exposure has a significant impact on the sleep,mood states,and brain function of populations from near-sea-level regions,and DLPFC,in particular,is closely associated with changes in sleep and mood states.The findings of this study provide a theoretical basis for health management and intervention strategies in high-altitude environments.

Objective:To investigate the clinical efficacy of vitamin D as an adjunct to low-dose aspirin on patients with early-onset severe preeclampsia.Methods:A retrospective analysis was conducted on the clinical data of 102 patients with early-onset severe preeclampsia admitted to Zhoushan Women and Children's Hospital from January 2020 to January 2024. Fifty patients admitted for treatment from January 2020 to January 2022 were included in the control group and received low-dose aspirin treatment. Fifty-two patients admitted between February 2022 and January 2024 were included in the observation group and received vitamin D as an adjunct to low-dose aspirin treatment. Both groups were treated for 7 days. Blood pressure, homocysteine, serum 25-hydroxyvitamin D 3, renal function indicators (glomerular filtration rate, 24-hour urine protein quantification), coagulation function indicators (prothrombin time, thrombin time, D-dimer, activated partial thromboplastin time), and pregnancy outcomes were compared between the two groups. Results:After treatment, systolic blood pressure, diastolic blood pressure, homocysteine, 24-hour urine protein quantification, and D-dimer in the observation group were (134.33 ± 6.28) mmHg (1 mmHg = 0.133 kPa), (88.57 ± 5.76) mmHg, (10.65 ± 3.15) μmol/L, (0.59 ± 0.31) g/24 hours, and (0.51 ± 0.32) mg/L, respectively. These values were significantly lower than those in the control group [(142.28 ± 6.04) mmHg, (93.85 ± 5.38) mmHg, (13.15 ± 2.89) μmol/L, (1.28 ± 0.47) g/24 hours, and (0.73 ± 0.49) mg/L, t = 4.17, -10.37, 4.17, 8.79, 2.69, all P < 0.05]. In the observation group, serum 25-hydroxyvitamin D 3 level and glomerular filtration rate were (27.94 ± 6.43) μg/L and (98.65 ± 14.72) mL·min?1·1.73 m?2, respectively. These values were significantly higher than those in the control group [(15.24 ± 5.92) μg/L, (90.19 ± 12.07) mL·min?1·1.73 m?2 ( t = -10.37, -3.18, both P < 0.05). Additionally, in the observation group, prothrombin time, thrombin time, and activated partial thromboplastin time were (12.19 ± 0.53) seconds, (12.19 ± 0.53) seconds, and (0.51 ± 0.32) seconds, respectively. These times were significantly shorter than those in the control group [(13.22 ± 1.15) seconds, (16.94 ± 2.07) seconds, and (34.21 ± 3.93) seconds ( t = 5.85, 4.27, 3.81, all P < 0.05). There was no statistically significant difference in incidence of pregnancy outcomes between the two groups ( P > 0.05). Conclusions:Vitamin D as an adjunct to low-dose aspirin for the treatment of early-onset severe preeclampsia significantly reduces blood pressure, enhances blood circulation, promotes metabolism, improves coagulation function, and aids in the recovery of renal function in patients.

Objective:Given the insufficiency of current clinical biomarkers for early diagnosis of esophageal adenocarcinoma(EAC),this study developed a panel of serum protein biomarker assays using liquid-phase chip technology and preliminarily validated the detection capabilities of these markers for EAC.Methods:Collected serum samples from 48 patients with EAC and 33 age-matched healthy controls(HC).The levels of squamous cell carcinoma antigen(SCCA),human cytokeratin 19 fragment(Cyfra21-1),hepatocyte growth factor(HGF)and IL-8 in serum were analyzed by liquid chip technology.The diagnostic efficacy of a single indicator and a combination of four indicators were evaluated by receiver operation characteristic(ROC)curve.Results:The detection ranges of SCCA,Cyfra21-1,HGF and IL-8 were 0.24～1 000 ng/ml,45.72～100 000 pg/ml,21.95～16 000 pg/ml,and 0.61～10 000 pg/ml,respectively.The liquid chip technology shows a strong correlation with clinical chemiluminescence immunoassay(r=0.949 4,P<0.000 1)and ELISA technology(r=0.955 1,P<0.000 1).Compared to the HC group,serum levels of SCCA and HGF were signifi-cantly elevated in the EAC group(P<0.01),IL-8 was significantly decreased(P<0.05),while Cyfra21-1 shows no significant difference(P>0.05).In the early EAC group,serum HGF was significantly higher than in the HC group(P<0.01).ROC curve analysis shows that among individual markers,HGF exhibits the best diagnostic efficacy for early EAC with an area under the curve(AUC)of 0.761,while the AUC for the combination of the four biomarkers was 0.857,both superior to the clinical biomarkers SCCA(AUC=0.604)and Cyfra21-1(AUC=0.515).Conclusion:Liquid chip technology is used to jointly detect SCCA,Cyfra21-1,HGF,and IL-8 in human serum.In the early diagnosis of EAC,the diagnostic efficacy of the combined use of these four biomarkers is superior to that of the commonly used clinical tumor markers SCCA and Cyfra21-1.This advantage stems from the high throughput and high sensitivity characteristics of liquid-phase chip technology.Consequently,this combined detection method holds significant clinical application and is expected to provide a more accurate and reliable tool for the early diagnosis of EAC.

The classification of stoma skin barriers varies based on their specific features. The curvature design of convex skin barriers provides a secure and effective seal for patients with flat, retracted stomas or peristomal skin folds. The secure sealing ability of convex skin barriers is attributed to several critical structural components. Although convex skin barriers offer many clinical advantages, there is currently no unified standard for measuring their characteristics, resulting in confusion among healthcare professionals when selecting stoma care products. To address this issue, the 2021 International Stoma Care Expert Meeting proposed the Expert Consensus on Characteristics of Convex Skin Barriers and Clinical Application, which clearly defines five essential properties and clinical application guidelines for convex barriers. However, as most consensus contributors are from Europe and North America, its applicability in Chinese healthcare settings may be limited. Therefore, this paper provides a detailed interpretation of the five characteristics and clinical application statements of convex skin barriers, aiming to offer practical guidance to clinical nurses in selecting appropriate convex products and managing stoma-related complications.

Objective:To investigate the clinical efficacy of vitamin D as an adjunct to low-dose aspirin on patients with early-onset severe preeclampsia.Methods:A retrospective analysis was conducted on the clinical data of 102 patients with early-onset severe preeclampsia admitted to Zhoushan Women and Children's Hospital from January 2020 to January 2024. Fifty patients admitted for treatment from January 2020 to January 2022 were included in the control group and received low-dose aspirin treatment. Fifty-two patients admitted between February 2022 and January 2024 were included in the observation group and received vitamin D as an adjunct to low-dose aspirin treatment. Both groups were treated for 7 days. Blood pressure, homocysteine, serum 25-hydroxyvitamin D 3, renal function indicators (glomerular filtration rate, 24-hour urine protein quantification), coagulation function indicators (prothrombin time, thrombin time, D-dimer, activated partial thromboplastin time), and pregnancy outcomes were compared between the two groups. Results:After treatment, systolic blood pressure, diastolic blood pressure, homocysteine, 24-hour urine protein quantification, and D-dimer in the observation group were (134.33 ± 6.28) mmHg (1 mmHg = 0.133 kPa), (88.57 ± 5.76) mmHg, (10.65 ± 3.15) μmol/L, (0.59 ± 0.31) g/24 hours, and (0.51 ± 0.32) mg/L, respectively. These values were significantly lower than those in the control group [(142.28 ± 6.04) mmHg, (93.85 ± 5.38) mmHg, (13.15 ± 2.89) μmol/L, (1.28 ± 0.47) g/24 hours, and (0.73 ± 0.49) mg/L, t = 4.17, -10.37, 4.17, 8.79, 2.69, all P < 0.05]. In the observation group, serum 25-hydroxyvitamin D 3 level and glomerular filtration rate were (27.94 ± 6.43) μg/L and (98.65 ± 14.72) mL·min?1·1.73 m?2, respectively. These values were significantly higher than those in the control group [(15.24 ± 5.92) μg/L, (90.19 ± 12.07) mL·min?1·1.73 m?2 ( t = -10.37, -3.18, both P < 0.05). Additionally, in the observation group, prothrombin time, thrombin time, and activated partial thromboplastin time were (12.19 ± 0.53) seconds, (12.19 ± 0.53) seconds, and (0.51 ± 0.32) seconds, respectively. These times were significantly shorter than those in the control group [(13.22 ± 1.15) seconds, (16.94 ± 2.07) seconds, and (34.21 ± 3.93) seconds ( t = 5.85, 4.27, 3.81, all P < 0.05). There was no statistically significant difference in incidence of pregnancy outcomes between the two groups ( P > 0.05). Conclusions:Vitamin D as an adjunct to low-dose aspirin for the treatment of early-onset severe preeclampsia significantly reduces blood pressure, enhances blood circulation, promotes metabolism, improves coagulation function, and aids in the recovery of renal function in patients.

Objective:Given the insufficiency of current clinical biomarkers for early diagnosis of esophageal adenocarcinoma(EAC),this study developed a panel of serum protein biomarker assays using liquid-phase chip technology and preliminarily validated the detection capabilities of these markers for EAC.Methods:Collected serum samples from 48 patients with EAC and 33 age-matched healthy controls(HC).The levels of squamous cell carcinoma antigen(SCCA),human cytokeratin 19 fragment(Cyfra21-1),hepatocyte growth factor(HGF)and IL-8 in serum were analyzed by liquid chip technology.The diagnostic efficacy of a single indicator and a combination of four indicators were evaluated by receiver operation characteristic(ROC)curve.Results:The detection ranges of SCCA,Cyfra21-1,HGF and IL-8 were 0.24～1 000 ng/ml,45.72～100 000 pg/ml,21.95～16 000 pg/ml,and 0.61～10 000 pg/ml,respectively.The liquid chip technology shows a strong correlation with clinical chemiluminescence immunoassay(r=0.949 4,P<0.000 1)and ELISA technology(r=0.955 1,P<0.000 1).Compared to the HC group,serum levels of SCCA and HGF were signifi-cantly elevated in the EAC group(P<0.01),IL-8 was significantly decreased(P<0.05),while Cyfra21-1 shows no significant difference(P>0.05).In the early EAC group,serum HGF was significantly higher than in the HC group(P<0.01).ROC curve analysis shows that among individual markers,HGF exhibits the best diagnostic efficacy for early EAC with an area under the curve(AUC)of 0.761,while the AUC for the combination of the four biomarkers was 0.857,both superior to the clinical biomarkers SCCA(AUC=0.604)and Cyfra21-1(AUC=0.515).Conclusion:Liquid chip technology is used to jointly detect SCCA,Cyfra21-1,HGF,and IL-8 in human serum.In the early diagnosis of EAC,the diagnostic efficacy of the combined use of these four biomarkers is superior to that of the commonly used clinical tumor markers SCCA and Cyfra21-1.This advantage stems from the high throughput and high sensitivity characteristics of liquid-phase chip technology.Consequently,this combined detection method holds significant clinical application and is expected to provide a more accurate and reliable tool for the early diagnosis of EAC.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.