Objective To investigate the conformational states of ADAMTS13 under different pH conditions.Methods Atomic force microscopy(AFM)was applied to pull ADAMTS13 molecules with two distinct pulling systems:the Biotin-Streptavidin system and the 6×His-Anti-His antibody system.The rupture forces and molecular contour lengths were analyzed.Results Under pH 7.4 condition,when ADAMTS13 was pulled by Biotin-Streptavidin system,the mean molecular contour length was(30.93±1.56)nm,exhibiting a bimodal frequency distribution with peak positions at(22.12±0.01)and(49.57±0.05)nm.When ADAMTS13 was pulled using 6×Hist-anti-His antibody system,the mean molecular contour length was(32.77±0.72)nm,also showing a bimodal distribution,with a peak position at(25.73±0.16)and(43.84±0.63)nm,respectively.Under pH 6.0 condition,when ADAMTS13 was pulled by Biotin-Streptavidin system,the mean molecular contour length increased to(47.07±1.6)nm,and the frequency distribution shifted to trimodal,with peak positions at(22±1.25),(55.09±2.62)and(76.69±3.06)nm.The conformation of ADAMTS13 was more extended at pH 6.0 compared with that at pH 7.4.Conclusions ADAMTS13 exists in'closed'and intermediate conformational states at physiological pH 7.4.However,at pH 6.0,ADAMTS13 can adopt'closed',intermediate,and'open'conformational states.This study contributes to a further understanding of the role of ADAMTS13 in normal physiology and thrombotic thrombocytopenic purpura,providing insights for the development of novel recombination ADAMTS13 drugs.

ObjectiveTo analyze the epidemiological characteristics and influencing factors of SARS-CoV-2 reinfection by conducting follow-up investigations among community residents who experienced their first SARS-CoV-2 infection between March and June 2022, so as to provide a scientific basis for predicting future epidemic trends and adjusting prevention and control strategies. MethodsA cohort study was conducted in Xuhui District, Shanghai. A total of 1 208 individuals with a documented primary SARS-CoV-2 infection between March and June 2022 were enrolled and followed-up longitudinally. Data were collected using structured questionnaire surveys to assess the reinfection rate, incidence density, and clinical manifestations of SARS-CoV-2 reinfection. A logistic regression model was used to analyze the influencing factors of SARS-CoV-2 reinfection. ResultsA total of 497 SARS-CoV-2 reinfection cases were observed among the 1 208 research subjects, with a reinfection rate of 41.14% and an incidence density of 0.63 cases per 1 000 person-days. The cumulative reinfection rates at 6, 9, 12, 15, and 18 months following the initial infection were 0.08%, 15.31%, 19.04%, 33.53%, and 38.25%, respectively. Compared with the primary infection, reinfection was more likely to be symptomatic, with a greater severity of fever, dry cough, sore throat, and runny nose. Being female, younger age, and symptom duration ≥7 days during the primary infection were identified as influencing factors for SARS-CoV-2 reinfection, while a higher socioeconomic status can reduce the risk of SARS-CoV-2 reinfection. ConclusionSARS-CoV-2 reinfection is relatively common and often symptomatic. Age, gender, income level, and the duration of symptoms during the primary infection are identified as infuencing factors for SARS-CoV-2 reinfection. Continuous monitoring of reinfection in the population is recommended, along with the development of effective strategies to mitigate the impact of reinfection.

Objective To investigate the conformational states of ADAMTS13 under different pH conditions.Methods Atomic force microscopy(AFM)was applied to pull ADAMTS13 molecules with two distinct pulling systems:the Biotin-Streptavidin system and the 6×His-Anti-His antibody system.The rupture forces and molecular contour lengths were analyzed.Results Under pH 7.4 condition,when ADAMTS13 was pulled by Biotin-Streptavidin system,the mean molecular contour length was(30.93±1.56)nm,exhibiting a bimodal frequency distribution with peak positions at(22.12±0.01)and(49.57±0.05)nm.When ADAMTS13 was pulled using 6×Hist-anti-His antibody system,the mean molecular contour length was(32.77±0.72)nm,also showing a bimodal distribution,with a peak position at(25.73±0.16)and(43.84±0.63)nm,respectively.Under pH 6.0 condition,when ADAMTS13 was pulled by Biotin-Streptavidin system,the mean molecular contour length increased to(47.07±1.6)nm,and the frequency distribution shifted to trimodal,with peak positions at(22±1.25),(55.09±2.62)and(76.69±3.06)nm.The conformation of ADAMTS13 was more extended at pH 6.0 compared with that at pH 7.4.Conclusions ADAMTS13 exists in'closed'and intermediate conformational states at physiological pH 7.4.However,at pH 6.0,ADAMTS13 can adopt'closed',intermediate,and'open'conformational states.This study contributes to a further understanding of the role of ADAMTS13 in normal physiology and thrombotic thrombocytopenic purpura,providing insights for the development of novel recombination ADAMTS13 drugs.

ObjectiveTo analyze the epidemiological characteristics of long COVID and to investigate its main influencing factors by examining individuals infected with SARS-CoV-2 between March and June 2022 in two communities in Shanghai, to lay the foundation for further research on the mechanism and clinical treatment of long COVID, and to provide the basis for the development of inexpensive, convenient, and feasible prevention and intervention strategies. MethodsA cross-sectional study was conducted, enrolling 6 410 individuals infected with SARS-CoV-2. Data were collected through a questionnaire survey. The incidence and common symptoms of long COVID were analyzed, along with their associations with demographic characteristics, medical history, and behavioral factors. A logistic regression model was used to identify the major factors associated with the development of long COVID symptoms. ResultsThe overall incidence rate of long COVID among the study population was 13.9%. The most commonly reported symptoms included fatigue (65.1%), attention disorders (23.1%), and cough (16.9%). The analysis showed that having underlying chronic diseases (OR=2.580, 95%CI: 2.165‒3.074), a history of allergies (OR=1.418, 95%CI: 1.003‒1.971), current smoking (OR=1.461, 95%CI: 1.013‒2.079), ever smoking (OR=2.462, 95%CI: 1.687‒3.551), a greater number of symptoms during the acute phase [1 symptom (OR=1.778, 95%CI: 1.459‒2.162), 2 symptoms (OR=2.749, 95%CI: 2.209‒3.409), ≥3 symptoms (OR=7.792, 95%CI: 6.333‒9.593)] and aggravated symptoms during the acute phase (OR=1.082, 95%CI: 1.070‒1.094) were factors associated with a higher risk of developing long COVID symptoms. Additionally, individuals who had consumed alcohol in the past year (OR=1.914, 95%CI: 1.344‒2.684) were more prone to objective long COVID symptoms. Among individuals under 50 years of age, females (OR=1.427, 95%CI: 1.052‒1.943) were more likely to develop objective long COVID symptoms. ConclusionThis study has identified the diversity of long COVID symptoms, which involve multiple organs and systems, including fatigue, attention disorders, cough, and joint pain. It has also revealed associations between long COVID and various demographic factors (e.g., age, gender), personal medical history (e.g., underlying chronic diseases, history of allergies), acute-phase characteristics (e.g., number and severity of symptoms), and behavioral factors (e.g., smoking, alcohol consumption). These findings highlight the need for further research and ongoing surveillance of long COVID and may inform the development of more targeted health management strategies for specific populations.

Objective:To explore the clinical manifestations, auxiliary examinations, treatment, and outcomes of autoimmune-associated epilepsy (AAE) with anti-glutamic acid decarboxylase 65 (GAD65) antibodies.Methods:Nine patients diagnosed with AAE with anti-GAD65 antibodies between July 2019 and December 2023 at the Department of Neurology of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, were enrolled. Their clinical data and prognosis were retrospectively analyzed.Results:The onset age of these 9 cases was 13.0-59.0 (29.0±13.4) years, with 8 females and 1 male. The main clinical phenotypes at onset included limbic encephalitis/extralimbic encephalitis (6 cases) and chronic epilepsy (3 cases, of whom 1 case was accompanied with stiff person syndrome). The former often demonstrated T 2/fluid attenuated inversion recovery (FLAIR) hyperintensities and enlargement of hippocampus and amygdala (5 cases), and may present with concomitant meningoencephalitis (2 cases) or hypertrophic pachymeningitis (1 case). The latter showed normal (2 cases) or increased T 2/FLAIR signals in the hippocampal tails (1 case) in magnetic resonance imaging. Electroencephalogram findings were usually characterized by slow activities (6 cases) and temporal epileptiform discharges (6 cases). Five cases concurrently presented with one or more other autoimmune diseases, including the autoimmune polyendocrine syndrome type Ⅱ in 1 case. All cases received first-line immunotherapy and 7 cases received long-term immunosuppressive agents. Eight cases were followed up for 5-45 months and 2 cases experienced relapse of autoimmune encephalitis (presented with new-onset seizures). At the last follow-up, all cases achieved favourable outcomes according to the modified Rankin Scale scores (all≤2) and 4 patients had reduced seizures, of whom 2 were seizure-free. Conclusions:Anti-GAD65 antibody-positive AAE is mainly composed of postencephalitic epilepsy and chronic temporal lobe epilepsy. The co-occurrence of systemic autoimmune diseases is common, and meningeal lesions sometimes can be involved in MRI findings. Patients usually have poor response to immunotherapy and anti-seizure medication,and some postencephalitic epilepsy patients who receive early immunotherapy and long-term immunosuppressants can achieve sustained seizure freedom.

The World Health Organization (WHO) classification serves as the internationally recognized standard for diagnosing and classifying hematopoietic and lymphoid tissue tumors(WHO-HEAM). Since 2001, it has undergone multiple upgrades and revisions, updating, clarifying, and refining previous tumor diagnostic and classification standards while incorporating numerous new genetic and molecular biological subtypes. In 2022, two classification proposals emerged due to a wealth of clinical and scientific research results: the fifth edition of the WHO hematopoietic and lymphoid tissue classification (WHO-HAEM5), published in Leukemia journal; and the International Consensus Classification (ICC), published in Blood journal. These two schemes differ in their approach to classifying hematopoietic and lymphoid tissue tumors, posing challenges for clinical laboratory diagnosis and treatment.

Objective:To explore the association between insulin resistance (IR) and genome-wide DNA methylation based on Shanghai twin study.Methods:Monozygotic twins (MZ) from Shanghai were recruited during 2012-2013, 2017-2018, and 2022-2023. Data were collected by questionnaire survey, physical examination and laboratory tests. Genome-wide DNA methylation was quantified. Generalized linear mixed effect model was applied to analyze the association between methylation level at each site and homeostatic model assessment 2-insulin resistance (HOMA2-IR). Non-paired and paired designs were used to assess the association between DNA methylation and phenotype of IR. Cluster analysis was conducted to identify the clusters of top significant sites. Generalized linear regression was performed to examine the differential methylation patterns from clusters.Results:A total of 100 MZ pairs were included in this study. Hypermethylated cg10535199-2q23.1 ( β=0.74%, P=1.51×10 -7, OR=1.06, 95% CI: 1.03-1.09) and ch.17.49619327- SPOP ( β=0.23%, P=7.54×10 -7, OR=1.17, 95% CI: 1.08-1.28) were identified with suggestive significance. After correcting for multiple testing, no sites reached genome-wide significance. There was no statistical significance in the paired analysis. Two clusters with hypomethylated ( β=-0.39%, P<0.001) and hypermethylated ( β=0.47%, P<0.001) patterns were observed for HOMA2-IR. Conclusions:IR was significantly associated with DNA methylation, and genetic factors might contribute to the association.

Objective To compare and explore the differences between the eight batches of drugs in the centralized procurement list and the 2018 edition of the national essential medicine list,and to provide reference for updating and improving the national essential medicine list and the national centralized procurement list of drugs.Methods The category,generic name variety,specification,and other information of drugs included in the centralized drug procurement were collected and compared with the 2018 edition of the national essential medicine list,and the reasons for differences were analyzed.Results A proportion of 39%of centralized procurement drugs were listed in national essential medicines.Forty pharmacological classifications were not involved in the drugs of centralized procurement.Only anticoagulant and thrombolytic drugs with dual attributes accounted for a smaller variety proportion than the specification proportion.Conclusion There are some differences between the centralized procurement list and the 2018 edition of the national essential medicine list,which have some rationality,but also some problems to be solved.

Objective To explore the current status of medication literacy among urban elderly patients with chronic diseases in Anhui Province,aiming to reveal the factors influencing their medication literacy,and to propose targeted measures for improvement.Methods This research involved 381 participants aged 60 and above.It was conducted in Anhui province between December,2022 and January,2023,with data collected through face-to-face interviews by pharmacists.Single-factor analysis and ordinal multi-class logistic regression analysis were conducted to determine factors affecting medication literacy.Results Medication literacy cognition and medication literacy behavior were rated as good among urban older adults in Anhui province of the 294 valid questionnaires.Those who did not understood package insert exhibited significantly lower medication literacy behavior than those who fully understood[estimate=-1.224,95%CI=(-2.130,-0.317),P<0.01].Elderly patients with chronic diseases faced issues such as an inability to read or understand drug instructions in the investigation.90.48%of elderly patients with chronic diseases never heard or seldom heard of medication guidance services.Conclusion Medication literacy among urban elderly patients with chronic diseases is generally good in Anhui province.The ability to understood drug instructions significantly influenced the medication literacy of urban elderly patients with chronic diseases.Modifying the drug instructions to meet the reading needs of the elderly patients with chronic diseases and developing pharmaceutical care could effectively enhance rational drug use among this demographic.

Objective:To study the role and specific mechanisms of glucose-6-phosphate dehydrogenase (G6PD), a key enzyme for glycometabolism, mediated reduction stress in arsenic-induced malignant transformation of cells.Methods:Immortalized human skin keratinocyte-forming cells (HaCaT cells) were treated with sodium arsenite (NaAsO 2) at a concentration of 0.0 (control group) and 1.0 μmol/L (arsenic group), and malignant transformation indicators were tested using cell growth kinetics assay, cell scratch assay, and soft agar colony formation assay. At different stages of arsenic treatment (0, 1, 7, 14, 21, 28, 35 passages of cells), the effects of NaAsO 2 on glycometabolism in HaCaT cells were determined using corresponding reagent kits and Western blot, including glucose-6-phosphate (G6P), lactate, acetyl CoA, G6PD levels, as well as protein expression levels of hexokinase 2 (HK-2), 6-phosphofructose-2-kinase/fructose-2, 6-diphosphatase 3 (PFKFB3), pyruvate dehydrogenase kinase 1 (PDK1), 6-phosphoglucose dehydrogenase (PGD), and G6PD. Mitochondria were extracted, and the effects of NaAsO 2 on HaCaT cells and mitochondrial redox [reduced nicotinamide adenine dinucleotide phosphate (NADPH)/nicotinamide adenine dinucleotide phosphate (NADP +) ratio, reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio] were determined using corresponding reagent kits. The effect of G6PD on reduction stress and NaAsO 2-induced malignant transformation of HaCaT cells was determined using small interfering RNA (siRNA) intervention method. Results:Compared 1.0 μmol/L NaAsO 2-cultured HaCaT cells up to 35 generations (T-HaCaT cells) with matching passage 0.0 μmol/L NaAsO 2-cultured HaCaT cells [(33.797 ± 0.280) h, 0.177 ± 0.015, 13.667 ± 2.625], the multiplication time [(24.042 ± 0.479) h] was shorter ( t = 30.45, P < 0.001), the cell migration rate (0.396 ± 0.039) was higher ( t = 9.08, P < 0.001), and the number of colonies formed in soft agar (73.667 ± 4.450) was higher ( t = 20.11, P < 0.001). Compared with matching passage control group cells and 0 generation of the same group, G6P level in the arsenic group was higher at passages 1, 7, 14, 21, 28 and 35 ( P < 0.05), lactate and G6PD levels were higher at passages 14, 21, 28 and 35 ( P < 0.05), acetyl CoA level was lower at passages 21, 28 and 35 ( P < 0.05), and protein expression levels of HK-2, PFKFB3, PDK1, PGD, and G6PD were higher at passages 7, 14, 21, 28 and 35 ( P < 0.05). The NADPH/NADP + ratio of cells was higher at passages 1, 14, 21, 28 and 35 ( P < 0.05), GSH/GSSG ratio was higher at passages 21, 28 and 35 ( P < 0.05). The ratio of mitochondrial NADPH/NADP + was higher at passages 1, 7, 21, 28 and 35 ( P < 0.05), the GSH/GSSG ratio was higher at passages 1, 7, 14, 21, 28 and 35 ( P < 0.05). G6PD expression was silenced by siRNA in T-HaCaT cells, compared with the T-HaCaT con siRNA-treated group, the T-HaCaT G6PD siRNA-treated group had lower NADPH/NADP + and GSH/GSSG ratios in both cells and mitochondria ( P < 0.05), longer cell multiplication time, lower cell migration rate, and fewer soft agar colonies ( P < 0.05). Conclusion:During the malignant transformation of HaCaT cells induced by NaAsO 2, G6PD and other enzymes related to glycometabolism are activated, leading to reprogramming of glycometabolism, resulting in an imbalance of cell redox homeostasis and enhanced reduction stress in cells and mitochondria, thereby promoting NaAsO 2 induced malignant transformation of HaCaT cells.