OBJECTIVE: CRISPR-Cas protects bacteria from exogenous DNA invasion and is associated with bacterial biofilm formation and pathogenicity. METHODS: We analyzed the type I-F CRISPR-Cas system of Legionella pneumophila WX48, including Cas1, Cas2-Cas3, Csy1, Csy2, Csy3, and Cas6f, along with downstream CRISPR arrays. We explored the effects of the CRISPR-Cas system on the in vitro growth, biofilm-forming ability, and pathogenicity of L. pneumophila through constructing gene deletion mutants. RESULTS: The type I-F CRISPR-Cas system did not affect the in vitro growth of wild-type or mutant strains. The biofilm formation and intracellular proliferation of the mutant strains were weaker than those of the wild type owing to the regulation of type IV pili and Dot/Icm type IV secretion systems. In particular, Cas6f deletion strongly inhibited these processes. CONCLUSION The type I-F CRISPR-Cas system may reduce biofilm formation and intracellular proliferation in L. pneumophila.
Legionella pneumophila/pathogenicity* ; CRISPR-Cas Systems ; Biofilms/growth & development* ; Phenotype ; Bacterial Proteins/metabolism* ; Gene Deletion

Solid-phase extraction(SPE)combined with surface-enhanced Raman spectroscopy(SERS)has emerged as a promising analytical technique for detection and analysis of trace components in complex sample matrices.SPE enriches analytes through selective adsorption and solvent elution,effectively increasing the concentration and signal intensity.SERS enables ultra-sensitive and highly selective molecular analysis through the use of SERS-active substrates engineered to amplify Raman signals.The integration of these two techniques overcomes the limitations of conventional Raman spectroscopy in low-concentration detection field,while significantly improving sample preparation efficiency and analytical accuracy.This review provided a comprehensive overview of the characteristics of three SPE-SERS coupling modes,including two-step,one-step,and online integration.Special emphasis was placed on recent advancements in one-step SPE-SERS approaches based on novel functional adsorbent materials such as graphene,metal-organic frameworks,covalent organic frameworks,and molecularly imprinted polymers.Furthermore,future directions and development prospects of SPE-SERS technology were discussed.

OBJECTIVE: To examine the effectiveness of Chinese medicine (CM) Lianhua Qingwen Granule (LHQW) and Jingyin Gubiao Prescription (JYGB) in asymptomatic or mild patients with Omicron infection in the shelter hospital. METHODS: This single-center retrospective cohort study was conducted in the largest shelter hospital in Shanghai, China, from April 10, 2022 to May 30, 2022. A total of 56,244 asymptomatic and mild Omicron cases were included and divided into 4 groups, i.e., non-administration group (23,702 cases), LHQW group (11,576 cases), JYGB group (12,112 cases), and dual combination of LHQW and JYGB group (8,854 cases). The length of stay (LOS) in the hospital was used to assess the effectiveness of LHQW and JYGB treatment on Omicron infection. RESULTS: Patients aged 41-60 years, with nadir threshold cycle (C_T) value of N gene <25, or those fully vaccinated preferred to receive CM therapy. Before or after propensity score matching (PSM), the multiple linear regression showed that LHQW and JYGB treatment were independent influence factors of LOS (both P<0.001). After PSM, there were significant differences in LOS between the LHQW/JYGB combination and the other groups (P<0.01). The results of factorial design ANOVA proved that the LHQW/JYGB combination therapy synergistically shortened LOS (P=0.032). CONCLUSIONS Patients with a nadir C_T value <25 were more likely to accept CM. The LHQW/JYGB combination therapy could shorten the LOS of Omicron-infected individuals in an isolated environment.
Humans ; Drugs, Chinese Herbal/therapeutic use* ; Male ; Female ; Middle Aged ; Adult ; China/epidemiology* ; Hospitalization ; COVID-19 Drug Treatment ; COVID-19/epidemiology* ; SARS-CoV-2 ; Retrospective Studies ; Treatment Outcome ; Length of Stay ; Young Adult ; Aged

BACKGROUND: Generalized pustular psoriasis (GPP), a rare and recurrent autoinflammatory disease, imposes a substantial burden on patients and society. Awareness of GPP in China remains limited. METHODS: This cross-sectional survey, conducted between September 2021 and May 2023 across 14 hospitals in China, included GPP patients of all ages and disease phases. Data collected encompassed demographics, clinical characteristics, economic impact, disease severity, quality of life, and treatment-related complications. Risk factors for GPP recurrence were analyzed. RESULTS: Among 127 patients (female/male ratio = 1.35:1), the mean age of disease onset was 25 years (1st quartile [Q1]-3rd quartile [Q3]: 11-44 years); 29.2% had experienced GPP for more than 10 years. Recurrence occurred in 75.6% of patients, and nearly half reported no identifiable triggers. Younger age at disease onset ( P  = 0.021) and transitioning to plaque psoriasis ( P  = 0.022) were associated with higher recurrence rates. The median diagnostic delay was 8 months (Q1-Q3: 2-41 months), and 32.3% of patients reported misdiagnoses. Comorbidities were present in 53.5% of patients, whereas 51.1% experienced systemic complications during treatment. Depression and anxiety affected 84.5% and 95.6% of patients, respectively. During GPP flares, the median Dermatology Life Quality Index score was 19.0 (Q1-Q3: 13.0-23.5). This score showed significant differences between patients with and without systemic symptoms; it demonstrated correlations with both depression and anxiety scores. Treatment costs caused financial hardship in 55.9% of patients, underscoring the burden associated with GPP. CONCLUSIONS The substantial disease and economic burdens among Chinese GPP patients warrant increased attention. Patients with early onset disease and those transitioning to plaque psoriasis require targeted interventions to mitigate the high recurrence risk.
Humans ; Male ; Female ; Psoriasis/pathology* ; Adult ; Cross-Sectional Studies ; Adolescent ; Child ; Young Adult ; Quality of Life ; Middle Aged ; China/epidemiology* ; Recurrence ; Risk Factors ; Surveys and Questionnaires ; East Asian People

Objective:To develop a maternal assessment scale integrating both positive (affirmation, optimism, self-confidence) and negative psychological states (fear, anxiety, depression) throughout the entire pregnancy cycle and evaluate its reliability and validity.Methods:In December 2020, the scale items were preliminarily identified through a literature review, forming a 55-item questionnaire for pilot survey and expert interviews. A pilot survey was conducted among registered pregnant women at the Obstetrics and Gynecology Hospital of Fudan University from April to May 2021. The feasibility and reliability of the questionnaire was validated through reliability and validity analysis, and revisions were made based on the feedback. The finalized version comprised 43 items, categorized into four key event dimensions (pregnancy, childbirth, transition to motherhood, and complications) and six psychological state dimensions (affirmation, fear, anxiety, depression, optimism, and self-confidence). Among these psychological states, affirmation, self-confidence, and optimism represent positive states, while fear, anxiety, and depression reflect negative states. A formal survey was conducted from December 2021 to November 2022. The normality, multicollinearity, reliability, construct validity, convergent validity, and discriminant validity of each item were analyzed.Results:(1) General information: A total of 625 participants were involved in the pilot survey. For the formal survey, 8 045 questionnaires were distributed, with 6 273 valid responses (78.0%). Among the valid questionnaires, 5 694 (90.8%) reported positive psychological states and 579 (9.2%) negative states. All of the psychological state dimensions were correlated (all P<0.01), with no multicollinearity detected [variance inflation factor (VIF)<10]. The four key event dimensions were also correlated (all P<0.01), with no multicollinearity (VIF<10). (2) Reliability: The overall Cronbach's α coefficient of the questionnaire was 0.830, and removing any single item resulted in the value remaining>0.6. Cronbach's α coefficient values for affirmation, fear, anxiety, depression, optimism, and self-confidence were 0.772, 0.724, 0.648, 0.551, 0.257, and 0.740, respectively. The values for the key event dimensions were as follows: 0.722 for pregnancy, 0.554 for childbirth, 0.621 for transition to motherhood, and 0.568 for complications. (3) Model fit: For the psychological states, the Chi-square to degrees of freedom ratio ( χ2/df) was 19.979 (>3), and the root mean square error of approximation (RMSEA) was 0.055 (<0.08). The model of key event dimensions had a χ2/df of 48.557, RMSEA of 0.087, comparative fit index of 0.400 (<0.9), and incremental fit index of 0.400 (<0.9). (4) Convergent and discriminant validity: The average variance extraction (AVE) values for affirmation, fear, anxiety, depression, optimism, and self-confidence were 0.407, 0.099, 0.188, 0.223, 0.419, and 0.362, with composite reliability (CR) values of 0.822, 0.730, 0.655, 0.584, 0.627, and 0.786, respectively. In the model of key event dimensions, the AVE values for pregnancy, childbirth, transition to motherhood, and complications were 0.167, 0.287, 0.328, and 0.166, with CR values of 0.555, 0.832, 0.746, and 0.633, respectively. Significant correlations were observed between all psychological dimensions except optimism-depression and self-confidence-anxiety pairs (all P<0.05). All four key event dimensions were significantly correlated (all P<0.05). Conclusions:This study preliminarily develops a maternal psychological status assessment scale with satisfactory reliability and validity. This scale can be used to evaluate the comprehensive psychological states of pregnant women during critical pregnancy-related events.

Objective:To investigate the current situation of water-borne endemic fluorosis in Anhui Province, and provide basic data for the adjusting the prevention and control measures.Methods:Using cross-sectional survey method, all villages in the water-borne endemic fluorosis areas were investigated in Anhui Province from 2019 to 2022. In water-borne endemic fluorosis village, the situation of water improvement project and the fluoride level of drinking water were investigated, and dental fluorosis of all children aged 8 - 12 was examined. The criteria for determining the achievement of control targets for water-borne endemic fluorosis in affected counties were based on the "Evaluation Measures for Control and Elimination of Key Endemic Diseases (2019 Edition)".Results:From 2019 to 2022, the rate of water improvement village in water-borne endemic fluorosis areas were 88.47% (1 527/1 726), 100% (1 726/1 726), 100% (1 726/1 726) and 100% (1 726/1 726), respectively. The qualified proportion of water fluoride in water-borne endemic fluorosis villages was 33.84% (584/1 726), 63.09% (1 089/1 726), 70.74% (1 221/1 726) and 74.33% (1 283/1 726), respectively. The prevalence rates of dental fluorosis in children aged 8 - 12 were 25.48% (45 461/178 440), 15.78% (27 959/177 200), 13.68% (23 505/171 880) and 12.66% (23 315/184 200), respectively. The proportion of affected counties that had achieved the control target of water-borne endemic fluorosis was 16% (4/25), 60% (15/25), 36% (9/25) and 40% (10/25), respectively.Conclusions:The water-borne endemic fluorosis areas in Anhui Province have improved the water fluoride qualification rate and reduced the incidence of fluorosis in children through prevention and control measures such as water improvement and fluoride reduction. However, the prevention and control efforts in key areas and counties need to be further improved.

Objective:To investigate the current situation of water-borne endemic fluorosis in Anhui Province, and provide basic data for the adjusting the prevention and control measures.Methods:Using cross-sectional survey method, all villages in the water-borne endemic fluorosis areas were investigated in Anhui Province from 2019 to 2022. In water-borne endemic fluorosis village, the situation of water improvement project and the fluoride level of drinking water were investigated, and dental fluorosis of all children aged 8 - 12 was examined. The criteria for determining the achievement of control targets for water-borne endemic fluorosis in affected counties were based on the "Evaluation Measures for Control and Elimination of Key Endemic Diseases (2019 Edition)".Results:From 2019 to 2022, the rate of water improvement village in water-borne endemic fluorosis areas were 88.47% (1 527/1 726), 100% (1 726/1 726), 100% (1 726/1 726) and 100% (1 726/1 726), respectively. The qualified proportion of water fluoride in water-borne endemic fluorosis villages was 33.84% (584/1 726), 63.09% (1 089/1 726), 70.74% (1 221/1 726) and 74.33% (1 283/1 726), respectively. The prevalence rates of dental fluorosis in children aged 8 - 12 were 25.48% (45 461/178 440), 15.78% (27 959/177 200), 13.68% (23 505/171 880) and 12.66% (23 315/184 200), respectively. The proportion of affected counties that had achieved the control target of water-borne endemic fluorosis was 16% (4/25), 60% (15/25), 36% (9/25) and 40% (10/25), respectively.Conclusions:The water-borne endemic fluorosis areas in Anhui Province have improved the water fluoride qualification rate and reduced the incidence of fluorosis in children through prevention and control measures such as water improvement and fluoride reduction. However, the prevention and control efforts in key areas and counties need to be further improved.

Objective:To develop a maternal assessment scale integrating both positive (affirmation, optimism, self-confidence) and negative psychological states (fear, anxiety, depression) throughout the entire pregnancy cycle and evaluate its reliability and validity.Methods:In December 2020, the scale items were preliminarily identified through a literature review, forming a 55-item questionnaire for pilot survey and expert interviews. A pilot survey was conducted among registered pregnant women at the Obstetrics and Gynecology Hospital of Fudan University from April to May 2021. The feasibility and reliability of the questionnaire was validated through reliability and validity analysis, and revisions were made based on the feedback. The finalized version comprised 43 items, categorized into four key event dimensions (pregnancy, childbirth, transition to motherhood, and complications) and six psychological state dimensions (affirmation, fear, anxiety, depression, optimism, and self-confidence). Among these psychological states, affirmation, self-confidence, and optimism represent positive states, while fear, anxiety, and depression reflect negative states. A formal survey was conducted from December 2021 to November 2022. The normality, multicollinearity, reliability, construct validity, convergent validity, and discriminant validity of each item were analyzed.Results:(1) General information: A total of 625 participants were involved in the pilot survey. For the formal survey, 8 045 questionnaires were distributed, with 6 273 valid responses (78.0%). Among the valid questionnaires, 5 694 (90.8%) reported positive psychological states and 579 (9.2%) negative states. All of the psychological state dimensions were correlated (all P<0.01), with no multicollinearity detected [variance inflation factor (VIF)<10]. The four key event dimensions were also correlated (all P<0.01), with no multicollinearity (VIF<10). (2) Reliability: The overall Cronbach's α coefficient of the questionnaire was 0.830, and removing any single item resulted in the value remaining>0.6. Cronbach's α coefficient values for affirmation, fear, anxiety, depression, optimism, and self-confidence were 0.772, 0.724, 0.648, 0.551, 0.257, and 0.740, respectively. The values for the key event dimensions were as follows: 0.722 for pregnancy, 0.554 for childbirth, 0.621 for transition to motherhood, and 0.568 for complications. (3) Model fit: For the psychological states, the Chi-square to degrees of freedom ratio ( χ2/df) was 19.979 (>3), and the root mean square error of approximation (RMSEA) was 0.055 (<0.08). The model of key event dimensions had a χ2/df of 48.557, RMSEA of 0.087, comparative fit index of 0.400 (<0.9), and incremental fit index of 0.400 (<0.9). (4) Convergent and discriminant validity: The average variance extraction (AVE) values for affirmation, fear, anxiety, depression, optimism, and self-confidence were 0.407, 0.099, 0.188, 0.223, 0.419, and 0.362, with composite reliability (CR) values of 0.822, 0.730, 0.655, 0.584, 0.627, and 0.786, respectively. In the model of key event dimensions, the AVE values for pregnancy, childbirth, transition to motherhood, and complications were 0.167, 0.287, 0.328, and 0.166, with CR values of 0.555, 0.832, 0.746, and 0.633, respectively. Significant correlations were observed between all psychological dimensions except optimism-depression and self-confidence-anxiety pairs (all P<0.05). All four key event dimensions were significantly correlated (all P<0.05). Conclusions:This study preliminarily develops a maternal psychological status assessment scale with satisfactory reliability and validity. This scale can be used to evaluate the comprehensive psychological states of pregnant women during critical pregnancy-related events.

Endo-beta-N-acetylglucosaminidase (ENGase) is widely distributed in various organisms. The first reported ENGase activity was detected in Diplococcus pneumoniae in 1971. The protein (Endo D) was purified and its peptide sequence was determined in 1974. Three ENGases (Endo F1-F3) were discovered in Flavobacterium meningosepticum from 1982 to 1993. After that, the activity was detected from different species of bacteria, yeast, fungal, plant, mice, human, etc. Multiple ENGases were detected in some species, such as Arabidopsis thaliana and Trichoderma atroviride. The first preliminary crystallographic analysis of ENGase was conducted in 1994. But to date, only a few ENGases structures have been obtained, and the structure of human ENGase is still missing. The currently identified ENGases were distributed in the GH18 or GH85 families in Carbohydrate-Active enZyme (CAZy) database. GH18 ENGase only has hydrolytic activity, but GH85 ENGase has both hydrolytic and transglycosylation activity. Although ENGases of the two families have similar (β/α)8-TIM barrel structures, the active sites are slightly different. ENGase is an effective tool for glycan detection andglycan editing. Biochemically, ENGase can specifically hydrolyze β‑1,4 glycosidic bond between the twoN-acetylglucosamines (GlcNAc) on core pentasaccharide presented on glycopeptides and/or glycoproteins. Different ENGases may have different substrate specificity. The hydrolysis products are oligosaccharide chains and a GlcNAc or glycopeptides or glycoproteins with a GlcNAc. Conditionally, it can use the two products to produce a new glycopeptides or glycoprotein. Although ENGase is a common presentation in cell, its biological function remains unclear. Accumulated evidences demonstrated that ENGase is a none essential gene for living and a key regulator for differentiation. No ENGase gene was detected in the genomes of Saccharomyces cerevisiae and three other yeast species. Its expression was extremely low in lung. As glycoproteins are not produced by prokaryotic cells, a role for nutrition and/or microbial-host interaction was predicted for bacterium produced enzymes. In the embryonic lethality phenotype of the Ngly1-deficient mice can be partially rescued by Engase knockout, suggesting down regulation of Engase might be a solution for stress induced adaptation. Potential impacts of ENGase regulation on health and disease were presented. Rabeprazole, a drug used for stomach pain as a proton inhibitor, was identified as an inhibitor for ENGase. ENGases have been applied in vitro to produce antibodies with a designated glycan. The two step reactions were achieved by a pair of ENGase dominated for hydrolysis of substrate glycoprotein and synthesis of new glycoprotein with a free glycan of designed structure, respectively. In addition, ENGase was also been used in cell surface glycan editing. New application scenarios and new detection methods for glycobiological engineering are quickly opened up by the two functions of ENGase, especially in antibody remodeling and antibody drug conjugates. The discovery, distribution, structure property, enzymatic characteristics and recent researches in topical model organisms of ENGase were reviewed in this paper. Possible biological functions and mechanisms of ENGase, including differentiation, digestion of glycoproteins for nutrition and stress responding were hypothesised. In addition, the role of ENGase in glycan editing and synthetic biology was discussed. We hope this paper may provide insights for ENGase research and lay a solid foundation for applied and translational glycomics.

ObjectiveTo reveal the influence of Jianchangbang characteristic processing method on the change process of volatile components and the processing mechanism of reducing toxicity and increasing efficiency of Magnoliae Officinalis Cortex（MOC） by studying the changes in the composition and content of volatile components during the processing of ginger processed Xingpo pieces. MethodSamples of raw products， ginger juice moisturized products and stir-fried and heap moisturized products of MOC were taken according to the set time points， and headspace gas chromatography-mass spectrometry（HS-GC-MS） was used to determine the contents of volatile components in the samples， and the relative content of each component was obtained by peak area normalization. Principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were performed on the sample data using SIMCA 14.1 software， and the differential components during the processing were screened with variable importance in the projection（VIP） value>1 as the indicator. ResultA total of 68 volatile components were identified in the samples， among which some of the chemical components with similar structures showed similar trends of changes， and there was also the phenomenon of interconversion between compounds. Compared with the raw products， the contents of 42 components in ginger juice moisturized products increased， while the contents of 25 components decreased， 19 components were unique， and 4 components were unique to the raw products. Compared with ginger juice moisturized products， MOC in the early stage of piling had three unique components， and the contents of 11 components such as cyclosativene and （+）-α-pinene increased， and the contents of 5 components such as tricyclic terpene and α-curcumene decreased， and ginger juice moisturized products had four unique components. Compared with the early stage of piling， in the later stage， the contents of 8 components such as （+）-α-pinene and camphene significantly increased， while the contents of 6 components such as linalool and α-selinene significantly decreased. During the processing of MOC， there were significant changes in the chemical composition of the samples before and after 20 days. The differences between ginger juice moistening and the early stage of piling， the early stage and the later stage of piling could be clearly distinguished. ConclusionDuring the preparation process of ginger processed Xingpo pieces， the addition of ginger juice can reduce the contents of stimulating components， and the contents of active components continue to increase in several stages， such as the addition of ginger juice， frying and heap moisturizing， the quality of the decoction pieces may change significantly at about 20 d of processing. This study can provide a research basis for exploring the processing mechanism of ginger processed Xingpo pieces.