This study aims to establish a high-performance size-exclusion chromatography (HPSEC) method for determining the content of the classical swine fever virus (CSFV) E2 protein and screen the optimal stabilizer to enhance the stability of this protein. The optimal detection conditions were determined by optimizing the composition of the mobile phase, and characteristic chromatographic peaks were identified by SDS-PAGE and Western blotting. The specificity, repeatability, precision, linearity, limit of detection (LOD), and limit of quantitation (LOQ) of the method were assessed. The method established was used to determine the content of CSFV E2 protein antigen and vaccine. Differential scanning fluorimetry (DSF) was employed to screen the buffer system, pH, and salt ion concentrations, and sugar, amino acid, and alcohol stabilizers were further screened. The results showed that using a 200 mmol/L phosphate buffer provided the best column efficiency. An antigen-specific chromatographic peak appeared at the retention time of 18 min, which was identified as the CSFV E2 protein by SDS-PAGE and Western blotting. The method exhibited high specificity for detecting the CSFV E2 protein, with no absorbance peak observed in the blank control. The relative standard deviation (RSD) of the peak area for six repeated injections of the CSFV E2 protein was 0.74%, indicating good repeatability of the method. The RSD for repeated detection of two different concentrations of CSFV E2 protein samples by different operators at different time points was less than 2%, suggesting good intermediate precision of the method. The peak area of the CSFV E2 protein was linearly related to its concentration, with the regression equation showing R² of 1.000. The LOD and LOQ of the method were 14.88 μg/mL and 29.75 μg/mL, respectively. Application of the developed method in the detection of three batches of CSFV E2 protein antigen and three batches of vaccine demonstrated results consistent with those from the bicinchoninic acid (BCA) assay, which meant that the method could accurately determine the content of CSFV E2 protein antigen and vaccine. The DSF method identified 50 mmol/L Tris-HCl at pH 8.0 as the optimal buffer, and the addition of sugar and alcohol stabilizers further improved the stability of the CSFV E2 protein. The HPSEC method established in this study is simple, fast, and exhibits good accuracy and repeatability, enabling precise measurement of the CSFV E2 protein content. It is expected to play a crucial role in the quality control of the CSFV E2 vaccine. Furthermore, the strategy for improving the CSFV E2 protein stability, identified through DSF screening, has significant implications for enhancing the stability of the CSFV E2 vaccine.
Classical Swine Fever Virus/chemistry* ; Chromatography, Gel/methods* ; Animals ; Swine ; Viral Envelope Proteins/immunology*

Objective To explore the distribution and drug resistance changes of pathogenic bacteria in adult acute myeloid leukemia(AML)with bloodstream infection,and to analyze risk factors of death of patients.Methods Changes of detection rate of pathogenic bacteria and drug resistance rate of main pathogenic bacteria of 85 patients with AML and bloodstream infection 30 months before confirmed diagnosis(pathogenic bacteria detected from January 2017 to June 2019)and 30 months after diagnosis(from July 2019 to December 2021)were compared.According to the prognosis at 6 months after bloodstream infection,patients were divided into the death group(33 cases)and the survival group(52 cases).Logistic regression analysis was used to analyze risk factors of death in patients with AML complicated with bloodstream infection.Results A total of 98 strains of pathogenic bacteria were detected in 85 patients with AML complicated with bloodstream infection,mainly gram-negative bacteria(65/98,66.33%),followed by Gram-positive bacteria(29/98,29.59%)and fungi(4/98,4.08%).The proportion of fungi(all were candida)detected in the last 30 months was more than that in the first 30 months(P<0.05).There were no significant differences in proportions of gram-negative bacteria and gram-positive bacteria and drug resistance rates of Escherichia coli and Staphylococcus aureus between the late 30 months and the first 30 months(P>0.05).Logistic regression analysis showed that the history of antibiotic use within 1 month before confirmed diagnosis and septic shock were independent risk factors for death in patients with AML complicated with bloodstream infection(P<0.05).Conclusion The main pathogens of adults with AML combined with bloodstream infection are gram-negative bacteria.However,candida infection rate has increased in recent years,and patients with antibiotic use before bloodstream infection and complicated with septic shock are prone to poor prognosis.

Objective To investigate the risk factors of infection of Pseudomonas aeruginosa combined with Elizabethkingia anophelis in patients with cerebral hemorrhage and the antimicrobial treatment plan.Methods Clinical pharmacists participated in the treatment of pulmonary infection caused by Pseudomonas aeruginosa combined with Elizabethkingia anophelis in a patient with cerebral hemorrhage.The risk factors of Elizabethkingia anophelis infection and antimicrobial treatment plan were analyzed by referring to literature and combining the patient's condition,medical history,drug use history and related examination results.Results Based on the infection site,the characteristics of mixed bacterial infection,and the metabolic/pharmacodynamic characteristics of antimicrobial agents,clinical pharmacists made drug recommendations for clinicians in the adjustment of anti-infection protocols,and patients'systemic infections were effectively controlled.Conclusion Elizabethkingia anophelis is a conditional pathogen with low virulence and is not easy to infect healthy people.When the patient's immunity is low,it is easy to transform into pathogenic bacteria,which should be paid attention to.

Objective:To summarize the clinical and prognostic features of children with opsoclonus-myoclonus-ataxia syndrome (OMAS).Methods:A total of 46 patients who met the diagnostic criteria of OMAS in the Department of Neurology, Beijing Children′s Hospital from June 2015 to June 2023 were retrospectively analyzed. Centralized online consultations or telephone visits were conducted between June and August 2023. The data of the children during hospitalization and follow-up were collected, including clinical manifestations, assistant examination, treatment and prognosis. According to the presence or absence of tumor, the patients were divided into two groups. The chi-square test or Mann-Whitney U test was used to compare the differences between the two groups. Univariate Logistic regression was used to analyze the factors related to OMAS recurrence and prognosis. Results:There were 46 patients, with 25 males and the onset age of 1.5 (1.2, 2.4) years. Twenty-six (57%) patients were diagnosed with neuroblastoma during the course of the disease, and no patients were categorized into the high-risk group. A total of 36 patients (78%) were followed up for≥6 months, and all of them were treated with first-line therapy with glucocorticoids, gammaglobulin and (or) adrenocorticotrophic hormone. Among the 36 patients, 9 patients (25%) were treated with second-line therapy for ≥3 months, including rituximab or cyclophosphamide, and 17 patients (47%) received chemotherapy related to neuroblastoma. At the follow-up time of 4.2 (2.2, 5.5) years, 10 patients (28%) had relapsed of OMAS. The Mitchell and Pike OMS rating scale score at the final follow-up was 0.5 (0, 2.0). Seven patients (19%) were mildly cognitively behind their peers and 6 patients (17%) were severely behind. Only 1 patient had tumor recurrence during follow-up. The history of vaccination or infection before onset was more common in the non-tumor group than in the tumor group (55%(11/20) vs. 23%(6/26), χ2=4.95, P=0.026). Myoclonus occurred more frequently in the non-tumor group (40%(8/20) vs. 4%(1/26), χ2=7.23, P=0.007) as the onset symptom. Univariate Logistic regression analysis showed that the tumor group had less recurrence ( OR=0.19 (0.04-0.93), P=0.041). The use of second-line therapy or chemotherapy within 6 months of the disease course had a better prognosis ( OR=11.64 (1.27-106.72), P=0.030). Conclusions:OMAS in children mostly starts in early childhood, and about half are combined with neuroblastoma. Neuroblastoma in combination with OMAS usually has a low risk classification and good prognosis. When comparing patients with OMAS with and without tumors, the latter have a more common infection or vaccination triggers, and myoclonus, as the onset symptom, is more common. Early addition of second-line therapy is associated with better prognosis in OMAS.

Objective To introduces drug treatment and individualized pharmaceutical care for a patient with dilated cardiomyopathy digoxin poisoning and provides ideas for pharmaceutical care.Methods The pharmacist used therapeutic drug management to analyze the course of drug treatment before and after hospitalization,combined with therapeutic drug monitoring and drug-gene detection,to analyze the causes of poisoning in digoxin from the perspectives of underlying diseases,polymor-phism,drug dosage,combination of drugs,physiological and other pathological factors,and to assist in clinical drug reformula-tion and optimization of drug treatment regimens.Results The clinician accepted the clinical pharmacist's suggestion.The pa-tient had a good prognosis,and digoxin poisoning did not occur in the later period.Conclusion This case provides a feasible treatment for dilated cardiomyopathy and other patients with digoxin intoxication;it can be used as a reference for the prevention and treatment of digoxin poisoning and provide a new direction for the development of hospital pharmaceutical care and pharma-ceutical professionals.

Objective To analyze the influencing of vascular endothelial growth factor (VEGF) gene polymorphism on success rate of embryo implantation in patients with thin endometrium. Methods A total of 100 patients with thin endometrium from the gynecological outpatient and inpatient departments of the hospital from June 2022 to December 2023 were selected as study group, and another 100 healthy adult women from the physical examination department in the same period were selected as control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine the single nucleotide polymorphism at four different loci of the VEGF gene. Allele-specific PCR (AS-PCR) was used to analyze the distribution of VEGF genotypes and allele frequencies at the corresponding loci in both groups. The study group underwent frozen-thawed embryo transfer (FET), and their pregnancy conditions were confirmed by ultrasound 30 to 35 days later. Based on pregnancy outcomes, the study group was divided into successful pregnancy group and failed pregnancy group, and VEGF gene polymorphism between the two groups was compared. Logistic regression analysis was conducted to investigate the influence of VEGF gene polymorphism on pregnancy outcomes in patients with thin endometrium. Receiver operating characteristic (ROC) curve was plotted to assess the predictive value of VEGF gene polymorphism for pregnancy failure in patients with thin endometrium. Results Compared with the AA+CA genotype at the -2578C/A locus, patients carrying the CC genotype had increased risk of developing thin endometrium by 1.412 times (95%CI, 1.004 to 1.985), and the allelic mutation C showed increased risk by 1.526 times (95%CI, 1.066 to 2.186). Similarly, compared with the GA+AA genotype at the -1154G/A locus, patients carrying the GG genotype had increased risk of developing thin endometrium by 1.467 times (95%CI, 1.011 to 2.128), and the allelic mutation G showed increased risk by 1.531 times (95%CI, 1.026 to 2.284). There were no significant differences in VEGF gene genotypes and allele distributions at the -460C/T and +936C/T loci between the two groups (P>0.05). Among the 100 patients with thin endometrium in the study group, 40 cases had successful pregnancies after FET. There were significant differences in the distribution of VEGF gene genotypes at the -2578C/A and -1154G/A loci between the successful pregnancy group and the failed pregnancy group (P < 0.05). Logistic regression analysis revealed that carrying the CC genotype at the -2578C/A locus and the GG genotype at the -1154G/A locus were risk factors for pregnancy outcomes in patients with thin endometrium (OR>1, P < 0.05). The ROC curve showed that the area under the curve (AUC) for predicting pregnancy failure in patients with thin endometrium based on VEGF gene polymorphisms at the -2578C/A and -1154G/A loci was 0.801(95%CI, 0.552 to 0.838). Conclusion Carrying the CC genotype at the -2578C/A locus and the GG genotype at the -1154G/A locus can increase the risk of developing thin endometrium and reduce the success rate of pregnancy in patients with thin endometrium. Moreover, VEGF gene polymorphisms at the -2578C/A and -1154G/A loci can effectively predict the risk of pregnancy failure in patients with thin endometrium.

Objective:To investigate the iodine nutritional status of pregnant women in Qingdao and the effect of prevention and treatment of iodine deficiency disorders (IDD), so as to provide a basis for residents to supplement iodine scientifically, and take targeted prevention measures and adjust intervention strategies.Methods:In accordance with the requirements of the "National Iodine Deficiency Disorders Surveillance Program (2016 edition)" and "Shandong Iodine Deficiency Disorders Surveillance Program", the cluster sampling method was adopted to select pregnant women from 10 districts (cities) in Qingdao from 2018 to 2020, to investigate their basic information and thyroid disease history. Meanwhile, household edible salt samples and random urine samples were collected to detect iodine content.Results:A total of 3 000 pregnant women were monitored from 2018 to 2020, the median age was 31 years, and the median gestational age was 18 weeks. There were significant differences in the distribution of age, gestational age, whether senile puerpera, and pregnancy in different years ( H/χ 2 = 29.35, 81.03, 65.62, 77.34, P < 0.001). The median salt iodine of edible salt ( n = 3 000) and iodized salt ( n = 2 700) in pregnant women's homes were 23.02 and 23.70 mg/kg, respectively. The qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt were 89.59% (2 419/2 700), 90.00% (2 700/3 000) and 80.63% (2 419/3 000). The comparison of qualified rate of iodized salt, coverage rate of iodized salt and consumption rate of qualified iodized salt among different years was statistically significant (χ 2 = 48.09, 36.62, 61.08, P < 0.001), the coverage rate of iodized salt and the consumption rate of qualified iodized salt showed a downward trend year by year (χ 2trent = 35.54, 29.50, P < 0.001). A total of 3 000 urine samples were collected from pregnant women and the median urinary iodine of pregnant women was 147.85 μg/L. The urinary iodine level in the third trimester was lower than that in the first and second trimesters ( P < 0.001). The urinary iodine level in the non elderly group was higher than that in the elderly group ( Z = - 6.66, P < 0.001). The urinary iodine level in the group without thyroid disease was higher than that in the group with thyroid disease ( Z = - 1.99, P = 0.047). The urinary iodine level in iodized salt group was higher than that in non-iodized salt group ( Z = - 2.42, P = 0.015). Conclusions:The iodine nutrition of pregnant women in Qingdao is generally at an insufficient level, and the risk of iodine deficiency is high, which needs attention. In recent years, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in Qingdao have shown a downward trend, and have failed to meet the requirements of national standards. In the future, we should strengthen the monitoring and health education of IDD in pregnant women.

Progressive functional deterioration in the cochlea is associated with age-related hearing loss (ARHL). However, the cellular and molecular basis underlying cochlear aging remains largely unknown. Here, we established a dynamic single-cell transcriptomic landscape of mouse cochlear aging, in which we characterized aging-associated transcriptomic changes in 27 different cochlear cell types across five different time points. Overall, our analysis pinpoints loss of proteostasis and elevated apoptosis as the hallmark features of cochlear aging, highlights unexpected age-related transcriptional fluctuations in intermediate cells localized in the stria vascularis (SV) and demonstrates that upregulation of endoplasmic reticulum (ER) chaperon protein HSP90AA1 mitigates ER stress-induced damages associated with aging. Our work suggests that targeting unfolded protein response pathways may help alleviate aging-related SV atrophy and hence delay the progression of ARHL.
Mice ; Animals ; Transcriptome ; Aging/metabolism* ; Cochlea ; Stria Vascularis ; Presbycusis

Double aortic arch is a congenital aortic arch malformation, which is caused by abnormal development of the fourth aortic arches during embryonic development.The patient with double aortic arch has no specific symptoms.It often encircles the trachea and esophagus.In severe cases, it causes dyspnea and dysphagia, and even endangers life.Currently, double aortic arch is diagnosed mainly by imaging techniques, such as echocardiography, computed tomography angiography, magnetic resonance angiography, cardiac catheterization.The main treatment is surgery.Therefore early diagnosis and treatment are very important to avoid tracheomalacia and improve the prognosis.This article will briefly review the research progress of embryonic development mechanism, classification, clinical diagnosis, treatment and prognosis of double aortic arch, aiming to improve the physician′s skills of diagnosis and treatment.

Objective:To assess the operation and usage conditions of all water improvement projects in Qingdao, master the water fluoride content and the prevalence of dental fluorosis in children in drinking-water-borne endemic fluorosis villages, evaluate the prevention and control effect of water improvement project.Methods:From April to October in 2020, all drinking-water-borne endemic fluorosis villages in 7 cities (districts) of Qingdao were selected to investigate the operation and usage conditions of water improvement project, water fluoride and prevalence of dental fluorosis of 8 to 12 years old children. The prevention and control effect of water improvement project on drinking-water-borne endemic fluorosis in Qingdao was evaluated according to the control contents and criteria of drinking-water-borne endemic fluorosis.Results:There were 1 146 drinking-water-borne endemic fluorosis villages, the water improved rate of villages was 100.00% (1 146/1 146), the rate of normal operation of water improvement project was 99.91% (1 145/1 146). There were 1 130 villages with qualified water fluoride and the qualified rate was 98.60% (1 130/1 146). A total of 23 473 children aged 8 - 12 years old were examined, and 631 children were detected with dental fluorosis. The detection rate of dental fluorosis was 2.69% (631/23 473), the dental fluorosis index was 0.048, and the prevalence was negative. The detection rate of dental fluorosis of children in villages with excessive water fluoride (7.03%, 40/569) was higher than that in villages with qualified water fluoride (2.58%, 591/22 904, χ 2 = 41.235, P < 0.01). Conclusion:The operation and usage conditions of water improvement projects in 7 drinking- water-borne endemic fluorosis cities (districts) in Qingdao are good, and the detection rate of dental fluorosis in children is low, which has reached the national control standard.