Objective　To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Listeria monocytogenes, and analyze the structure and homology of CRISPR loci. Methods Totally 34 strains of Listeria monocytogenes isolated in our laboratory were identified, PCR amplified and sequenced. The repeat sequence structure and spacer sequence homology in CRISPR loci were analyzed by bioinformatics software. Results A total of 7 CRISPR loci were detected in 34 strains. The mutation rate of the first 2 and last 2 bases of the Repeat sequence of CRISPR loci was higher, while the mutation rate of the middle part was lower. Seven CRISPR sites form eight CRISPR structural types, among which the Repeat sequences of CRISPR1 and CRISPR2 are relatively conserved, while the Repeat sequences of CRISPR1 and CRISPR5 can form dumbbell shaped secondary structures. The number of Spacer sequences contained in each CRISPR site ranges from 2 to 15, with an average of 2.43. The 136 Spacer sequences detected were not only homologous to Listeria plasmids and bacteriophages, but also homologous to uncultured virus sequences, staphylococcal bacteriophages, and Listeria innocua. The same CRISPR genotype did not show large-scale clustering, but some strains in the same year were in the same evolutionary cluster with close genetic relationships. Conclusion The CRISPR structure of Listeria monocytogenes in this study exhibits high specificity, and its homology with bacteriophages provides a theoretical basis for the application of bacteriophages in the control and prevention of Listeria monocytogenes.

Objective To explore the pathogenic mechanisms of Legionella pneumophila(L.pneumophila)infection inhibiting the fusion of endosome-lysosome fusion in mouse macrophages.Methods Twelve C57 mice were randomly divided into control group and L.pneumophila infection group(n=6 each).After anesthesia,an equal volume of physiological saline or L.pneumophila solution was administered nasally.Body weight changes were monitored for 3 consecutive days,and the lungs were extracted to assess injury.Hematoxylin and eosin(HE)staining and immunohistochemical staining were performed to observe the pathological characteristics of lung tissue in both groups.Transcriptome sequencing was utilized to analyze differentially expressed genes(DEGs)and associated signaling pathways in lung tissues.Mouse bone marrow macrophages(BMDMs)were isolated and co-cultured with L.pneumophila,with infection status confirmed by immunofluorescence staining.Transcriptome sequencing was employed to analyze DEGs and enriched related signaling pathways before and after infection.Core genes involved in the post-infection signaling pathway were identified,and the consistency of their mRNA expression levels in vivo and in vitro was verified using RT-qPCR.The expression of relevant proteins was detected by Western Blotting,and bacterial proliferation assays were conducted to evaluate the intracellular replication of L.pneumophila.Results Compared with control group,the body weight of mice in L.pneumophila infection group significantly decreased(P<0.001)on the second and third day post-infection.Edema and red hepatoid degeneration were observed in both left and right lung tissues,with lesion areas spreading from the hilum to the lung periphery.HE staining revealed increased inflammatory cell infiltration in the alveolar spaces,thickening of alveolar septa and increased fibrin exudation in L.pneumophila infection group.Immunohistochemistry results showed a significant increase in myeloperoxidase(MPO)activity in the lung tissue infected mice(P<0.001).Transcriptome sequencing identified 2550 DEGs,with 1444 up-regulated genes and 1106 down-regulated genes.KEGG enrichment analysis indicated that these DEGs were mainly involved in pathways related to tumor necrosis factor,rheumatoid arthritis,Rap1,PI3K-Ak,and phagosome pathways.Immunofluorescence results showed in vitro proliferation of L.pneumophila within mouse BMDMs.Transcriptome sequencing identified 2550 DEGs,including 1677 up-regulated genes and 873 down-regulated genes.KEGG enrichment analysis showed that enrichment in pathway related to transcription dysregulation in cancer,PI3K-Akt and phagosome pathways.Thirteen core genes,including tubulin β1(Tubb1),were identified from the overlap between mouse lung tissue and BMDMs.RT-qPCR results demonstrated a significant decrease in Tubb1 expression in both lung tissue and BMDMs infected with L.pneumophila(P<0.001).Western Blotting results revealed significant decreases in Rab7,Tubb1,and LAMP2 protein expression(P<0.05),and increases in iNOS and MPO expression(P<0.05).Intracellular proliferation experiments indicated that L.pneumophila gradually increased within BMDMs over time.Conclusion The potential mechanism of L.pneumophila infection in mouse macrophages involves the down-regulation of Rab7/Tubb1/LAMP2 which inhibits the endosome-lysosome fusion.

Objective To investigate the inhibitory effect of antimicrobial peptide WK-13-3D on the proliferation of triple negative breast cancer MDA-MB-231 cells and its potential mechanism.Methods The effect of antimicrobial peptide WK-13-3D at concentrations of 0,10,15,20,25,30,35,and 40 μmol/L on MDA-MB-231 cells proliferation was assessed using the CCK-8 assay.A pull-down assay was conducted to identify interacting proteins of antimicrobial peptide WK-13-3D with MDA-MB-231 cells.MDA-MB-231 cells were obtained and divided into the following groups:(1)control group and treatment groups with 10 and 20 μmol/L antimicrobial peptide WK-13-3D.Apoptosis was evaluated using flow cytometry and Western blotting was conducted to detect the expression change of heavy chain binding protein(BiP),protein kinase R-like endoplasmic reticulum kinase(PERK),eukaryotic translation initiation factor 2α(eIF2α),phosphorylated eIF2α(p-eIF2α),and Bax proteins within the cells.(2)Control group(transfected with no-load plasmid),si-BiP-592 group(transfected with si-BiP-592 interference plasmid)and si-BiP-592+WK-13-3D group(co-treated with si-BiP-592 interference plasmid and 10 μmol/L antimicrobial peptide WK-13-3D).Western blotting was used to detect the expression changes of BiP,PERK,eIF2α,p-eIF2α and Bax proteins.Twelve BALB/c mice were randomly divided into PBS group(n=4),taxol(TAX)group(n=4)and WK-13-3D group(n=4).All mice were subcutaneously injected with MDA-MB-231 cells to establish a triple-negative breast cancer transplant tumor model.WK-13-3D group received local injections of antimicrobial peptide WK-13-3D[200 mg/(kg·d)],TAX group was administered TAX intraperitoneally at the same dose[200 mg/(kg·d)],and PBS group was injected with an equivalent volume of PBS.Two weeks post-injection,the mice were killed,and the tumor weight and volume were measured and photographed.Immunohistochemistry staining was performed to evaluate the expressions of BiP and Ki-67 proteins in the tumor tissues.Results CCK-8 assay showed a gradual decrease in MDA-MB-231 cell survival rates with increasing concentrations of WK-13-3D,with an inhibitory concentration 50(IC50)of 19.82 μmol/L.The pull-down assay identified 268 interacting proteins of antimicrobial peptides and MDA-MB-231 cells,mainly including heavy-chain binding protein(BiP),heat shock protein 90 beta family member 1(HSP90B1),valerin-containing protein(VCP),heat shock cognate 71 kD protein(HSPA8).Compared with control group,treatment with 10 and 20 μmol/L antimicrobial peptide WK-13-3D significantly increased the apoptosis rate of MDA-MB-231 cells(P<0.05 or P<0.01),decreased BiP protein expression(P<0.05),and increased the expression levels of PERK,p-eIF2α,and Bax(P<0.05 or P<0.01),with no significant change in eIF2α protein expression(P>0.05).Compared with control group,si-BiP-592 group showed BiP protein expression significantly decreased(P<0.05),and the expression of PERK,p-eIF2α,and Bax proteins was significantly increased(P<0.05),with no significant change in eIF2α protein expression(P>0.05);Compared with si-BiP-592 group,si-BiP-592+WK-13-3D group showed a decrease in BiP protein expression(P<0.05)and an increase in PERK,p-eIF2α,and Bax protein expression(P<0.05 or P<0.01),with no significant change in eIF2α protein expression(P>0.05).Tumor volumes in mice treated with antimicrobial peptide WK-13-3D and TAX were significantly smaller than those in PBS group(P<0.05),and the immunohistochemical staining showed that the proportion of Ki-67 and BiP positive cells in tumor tissues of WK-13-3D treated mice was significantly lower than that in PBS group(P<0.01).Conclusion Antimicrobial peptide WK-13-3D could inhibit the proliferation of MDA-MB-231 cells and its mechanism may involve the activation of endoplasmic reticulum stress and the induction of cell apoptosis.

Objective To explore the characteristics of latent profiles of ego depletion in patients with heart failure,and analyze the impacts of these profiles on patients' self-management behavior.Methods 280 heart failure patients who were hospitalized in a tertiary grade A hospital were conveniently selected in Nanjing from April to October 2024 as study subjects.The investigation utilized a general information questionnaire,the Ego-Depletion Scale for Heart Failure Patients,the Self-Management Scale of Heart Failure Patients,and the Chronic Disease Self-Efficacy Scale.The latent profile categories of ego depletion was explored in heart failure patients.Univariate analysis and multiple Logistic regression analysis were used to investigate the influencing factors of different latent profile categories and the impact of each category on patients' self-management levels.Results The ego depletion of 255 heart failure patients could be classified into 3 latent profiles:self-adaptive low depletion group(35.29％),symptom distressed moderate depletion group(43.53％),and social life change high depletion group(21.18％).Multiple Logistic regression analysis showed that residence,primary caregiver,education level,New York Heart Association cardiac function grade,and self-efficacy score were factors influencing the potential profile of ego depletion in heart failure patients(P＜0.05).The comparison of self-management scores among patients in different latent profiles of ego depletion showed statistically significant differences(P＜0.05).Conclusion The level of ego depletion in patients with heart failure is high with significant heterogeneity.Medical staff should pay attention to the heterogeneity and influencing factors of ego depletion in patients with heart failure in time,and develop personalized comprehensive intervention strategies to alleviate ego depletion and improve their self-management levels.

Objective:To analyze the iodine nutrition status of pregnant women in Weihai City before and after (2022, 2023) the iodine deficiency disorders (IDD) intervention, evaluate the effectiveness of intervention measures, and provide a scientific basis for adjusting IDD prevention strategies in the city.Methods:In May 2023, the intervention action of IDD was carried out for pregnent women in Weihai City. In April 2022 and December 2023, respectively, a cross-sectional survey method was conducted in four districts (cities) of Weihai City. Each district (city) was divided into five areas (east, west, south, north, and central) each year, and one township (street) was selected from each areas. At least 20 pregnant women were selected from each township (street) as survey subjects, and their household salt samples and random urine samples were collected to measure salt iodine and urinary iodine concentrations, and to compare the test results.Results:A total of 922 household salt samples were tested, including 530 iodized salt samples and 469 qualified iodized salt samples. The median salt iodine was 24.00 mg/kg. The coverage rate of iodized salt, the qualified rate of iodized salt, and the consumption rate of qualified iodized salt were 57.48%, 88.49%, and 50.87%, respectively. No significant difference was observed in medians salt iodine between 2022 (24.26 mg/kg) and 2023 (24.00 mg/kg, Z = - 1.58, P = 0.113). However, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in 2023 werehigher than those in 2022 [73.72% (373/506) vs 37.74% (157/416), 65.22% (330/506) vs 33.41% (139/416), χ 2 = 120.90, 92.40, P < 0.001]. A total of 922 urine samples were collected from pregnant women, with a median urinary iodine of 135.90 μg/L. The median urinary iodine of pregnant women in 2022 was 113.55 μg/L. There was a statistically significant difference in medians urinary iodine among different regions ( H = 27.91, P < 0.001). The median urinary iodine of pregnant women in 2023 was 153.00 μg/L. There was no statistically significant difference in medians urinary iodine among different regions ( H = 3.33, P = 0.343). The medians urinary iodine of pregnant women in Huancui District, Wendeng District, and Rushan City in 2023 (156.60, 155.00, 140.85 μg/L) were higher than those in 2022 (93.60, 110.00, 110.70 μg/L), and the differences were statistically significant ( Z = - 5.44, - 4.92, - 5.99, P < 0.001). The median urinary iodine of pregnant women in Weihai City in 2023 showed a statistically significant difference compared to 2022 ( Z = - 7.62, P < 0.001). Conclusions:The IDD intervention measures in Weihai City have achieved good results, and the coverage of iodized salt, the consumption rate of qualified iodized salt, and the iodine nutrition level of pregnant women have improved. We should continue to implement intervention measures, improve the iodine nutrition level of key populations, and maintain a sustained elimination of IDD.

Patients with Noonan syndrome (NS) are born with normal or slightly lower body length and weight compared to the normal ranges. However, their height gradually falls behind that of the general population, leading to growth retardation and delayed puberty. In China, the incidence of short stature in patients with NS is approximately 65%. Short stature in these patients arises from multiple causes, including feeding difficulties in infancy, comorbidities such as congenital heart disease, genetic heterogeneity, and disorders of the growth hormone/insulin-like growth factor-1 axis. Growth hormone is commonly used to alleviate symptoms of short stature. This article reviews the growth and development patterns at different stages of NS, analyzes the causes of short stature, and summarizes the latest advances in treatment to provide new insights for the diagnosis and management of short stature in patients with NS.
Noonan Syndrome/complications* ; Humans ; Body Height ; Growth Disorders/therapy*

OBJECTIVES: To study genotype-phenotype correlations in children with FGFR3 variants and to improve clinical recognition of related disorders. METHODS: Clinical data of 95 patients aged 0-18 years harboring FGFR3 variants, confirmed by whole‑exome sequencing at Shanghai Children's Medical Center from January 2012 to December 2023, were retrospectively reviewed. Detailed phenotypic characterization was performed for 22 patients with achondroplasia (ACH) and 10 with hypochondroplasia (HCH). RESULTS: Among the 95 patients, 52 (55%) had ACH, 24 (25%) had HCH, 9 (9%) had thanatophoric dysplasia, 3 (3%) had syndromic skeletal dysplasia, 2 (2%) had severe achondroplasia with developmental delay and acanthosis nigricans, and 5 (5%) remained unclassified. A previously unreported FGFR3 variant, c.1663G>T, was identified. All 22 ACH patients presented with disproportionate short stature accompanied by limb dysplasia, commonly with macrocephaly, a depressed nasal bridge, bowed legs, and frontal bossing; complications were present in 17 (77%). The 10 HCH patients predominantly exhibited disproportionate short stature with limb dysplasia and depressed nasal bridge. CONCLUSIONS ACH is the most frequent phenotype associated with FGFR3 variants, and missense variants constitute the predominant variant type. The degree of FGFR3 activation appears to correlate with the clinical severity of skeletal dysplasia.
Humans ; Receptor, Fibroblast Growth Factor, Type 3/genetics* ; Child ; Male ; Child, Preschool ; Female ; Infant ; Adolescent ; Dwarfism/genetics* ; Achondroplasia/genetics* ; Lordosis/genetics* ; Infant, Newborn ; Retrospective Studies ; Genetic Association Studies ; Bone and Bones/abnormalities* ; Phenotype ; Limb Deformities, Congenital

OBJECTIVE: To investigate possible associations between physical function assessment scales, such as Short Physical Performance Battery (SPPB) and Berg Balance Scale (BBS), with all-cause mortality in acute decompensated heart failure (ADHF) patients. METHODS: A total of 108 ADHF patients were analyzed from October 2020 to October 2022, and followed up to May 2023. The association between baseline clinical characteristics and all-cause mortality was analyzed by univariate Cox regression analysis, while for SPPB and BBS, univariate Cox regression analysis was followed by receiver operating characteristic curves, in which the area under the curve represented their predictive accuracy for all-cause mortality. Incremental predictive values for both physical function assessments were measured by calculating net reclassification index and integrated discrimination improvement scores. Optimal cut-off value for BBS was then identified using restricted cubic spline plots, and survival differences below and above that cut-off were compared using Kaplan-Meier survival curves and the log-rank test. The clinical utility of BBS was measured using decision curve analysis. RESULTS: For baseline characteristics, age, female, blood urea nitrogen, as well as statins, angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, or angiotensin receptor-neprilysin inhibitors, were predictive for all-cause mortality for ADHF patients. With respect to SPPB and BBS, higher scores were associated with lower all-cause mortality rates for both assessments; similar area under the curves were measured for both (0.774 for SPPB and 0.776 for BBS). Furthermore, BBS ≤ 36.5 was associated with significantly higher mortality, which was still applicable even adjusting for confounding factors; BBS was also found to have great clinical utility under decision curve analysis. CONCLUSIONS BBS or SPPB could be used as tools to assess physical function in ageing ADHF patients, as well as prognosticate on all-cause mortality. Moreover, prioritizing the improvement of balance capabilities of ADHF patients in cardiac rehabilitation regimens could aid in lowering mortality risk.

OBJECTIVE: To evaluate the clinical efficacy and safety of Yangxue Qingnao Pills (YXQNP) combined with amlodipine in treating patients with grade 1 hypertension. METHODS: This is a multicenter, randomized, double-blind, and placebo-controlled study. Adult patients with grade 1 hypertension of blood deficiency and Gan (Liver)-yang hyperactivity syndrome were randomly divided into the treatment or the control groups at a 1:1 ratio. The treatment group received YXQNP and amlodipine besylate, while the control group received YXQNP's placebo and amlodipine besylate. The treatment duration lasted for 180 days. Outcomes assessed included changes in blood pressure, Chinese medicine (CM) syndrome scores, symptoms and target organ functions before and after treatment in both groups. Additionally, adverse events, such as nausea, vomiting, rash, itching, and diarrhea, were recorded in both groups. RESULTS: A total of 662 subjects were enrolled, of whom 608 (91.8%) completed the trial (306 in the treatment and 302 in the control groups). After 180 days of treatment, the standard deviations and coefficients of variation of systolic and diastolic blood pressure levels were lower in the treatment group compared with the control group. The improvement rates of dizziness, headache, insomnia, and waist soreness were significantly higher in the treatment group compared with the control group (P<0.05). After 30 days of treatment, the overall therapeutic effects on CM clinical syndromes were significantly increased in the treatment group as compared with the control group (P<0.05). After 180 days of treatment, brachial-ankle pulse wave velocity, ankle brachial index and albumin-to-creatinine ratio were improved in both groups, with no statistically significant differences (P>0.05). No serious treatment-related adverse events occurred during the study period. CONCLUSIONS Combination therapy of YXQNP with amlodipine significantly improved symptoms such as dizziness and headache, reduced blood pressure variability, and showed a trend toward lowering urinary microalbumin in hypertensive patients. These findings suggest that this regimen has good clinical efficacy and safety. (Registration No. ChiCTR1900022470).
Humans ; Amlodipine/adverse effects* ; Drugs, Chinese Herbal/adverse effects* ; Male ; Female ; Hypertension/complications* ; Middle Aged ; Treatment Outcome ; Drug Therapy, Combination ; Adult ; Blood Pressure/drug effects* ; Double-Blind Method ; Aged ; Antihypertensive Agents/adverse effects*