Generation and characterization of a nanobody against the avian influenza virus H7 subtype.

Avian influenza virus (AIV) H7N9 diseases have been recently reported, raising concerns about a potential pandemic. Thus, there is an urgent need for effective therapeutics for AIV H7N9 infections. Herein, camelid immunization and yeast two-hybrid techniques were used to identify potent neutralizing nanobodies (Nbs) targeting the H7 subtype hemagglutinin. First, we evaluated the binding specificity and hemagglutination inhibition activity of the screened Nbs against the H7 subtype hemagglutinin. Nb-Z77, with high hemagglutination inhibition activity was selected from the screened Nbs to optimize the yeast expression conditions and construct oligomeric forms of Nb-Z77 using various ligation methods. The oligomers Nb-Z77-DiGS, Nb-Z77-TriGS, Nb-Z77-Fc and Nb-Z77-Foldon were successfully constructed and expressed. Nb-Z77-DiGS and Nb-Z77-Foldon exhibited considerably greater activity than did Nb-Z77 against H7 subtype hemagglutinin, with median effective concentrations of 384.7 and 27.33 pM and binding affinity values of 213 and 5.21 pM, respectively. Nb-Z77-DiGS and Nb-Z77-Foldon completely inhibited the hemagglutination activity of the inactivated virus H7-Re1 at the lowest concentration of 0.938 µg/mL. This study screened a strain of Nb with high hemagglutination inhibition activity and enhanced its antiviral activity through oligomerization, which may have great potential for developing effective agents for the prevention, diagnosis, and treatment of AIV H7 subtype infection.

Prevalence and genomic-based antimicrobial resistance analysis of Avibacterium paragallinarum isolates in Guangdong Province, China.

Infectious coryza (IC) is an acute infectious respiratory disease in chickens that is caused by Avibacterium paragallinarum (A. paragallinarum). A. paragallinarum poses a significant threat to poultry health due to its virulence and multidrug resistance. This study isolated and identified 21 A. paragallinarum isolates from Guangdong between 2022 and 2023. Biochemical tests showed that 100% of A. paragallinarum isolates fermented glucose but did not ferment alginate and galactose, and only YZ18 was nicotinamide adenine dinucleotide independent. To determine the genetic relatedness between these isolates and NCBI reference strains, whole-genome-based phylogenetic analysis was employed. In addition, analysis of the 2,000 bp-length hmtp210 gene showed that the hmtp210 gene was strongly associated with A. paragallinarum serotypes. Meanwhile, a PCR assay for serotyping A. paragallinarum was developed based on the hmtp210 gene, this assay has high sensitivity and specificity. The antimicrobial susceptibility of isolates was assessed using the disk diffusion method. The antibiotic resistance genes of isolates were analyzed using the genomic method. Phenotypic resistance to ampicillin (95.2%), streptomycin (95.2%), methotrexate-sulfamethoxazole (90.5%), and tetracycline (85.7%) was most frequent among the isolates. All of the isolates exhibited resistance to multiple drugs, and furthermore, the isolates possessed a collective total of 14 genes associated with antibiotic resistance. This study will contribute to advancing our knowledge of A. paragallinarum antibiotic resistance and provide a scientific basis for the prophylaxis and treatment of IC, and the subsequent rational design of potential clinical therapeutics.

Development and application of an indirect ELISA for detection of antibodies against emerging atypical porcine pestivirus.

BACKGROUND: Atypical porcine pestivirus (APPV) is a newly discovered swine pestivirus, which can cause congenital tremor and high mortality in newborn piglets and subclinical infection in adult pigs, leading to significant impacts on the pig industry. Currently, there is no approved serological method to assess APPV infection status in pig farms. METHODS: In this study, the envelope glycoprotein E2 of APPV was highly expressed in suspension HEK293 cells, and further an indirect enzyme-linked immunosorbent assay based on the recombinant E2 protein (E2-iELISA) was developed and evaluated. RESULTS: The reaction parameters of the E2-iELISA were optimized, and the cutoff value was determined to be 0.2 by analyzing S/P values of 165 negative sera against APPV that were confirmed by virus neutralization test (VNT). Specificity test showed that the method had no cross-reaction with other common swine viruses. The E2-iELISA was evaluated using a panel of swine sera, and showed high sensitivity (113/120, 94.2%) and specificity (65/70, 92.9%), and the agreement rate with VNT was 93.7% (178/190). Subsequently, the E2-iELISA was utilized to investigate the seroprevalence of APPV in pig herds of China. When detecting 1368 pig serum samples collected from nine provinces in China, the overall seroprevalence of APPV was 73.9% (1011/1368). CONCLUSION: Our findings suggest that the E2-iELISA is specific and sensitive, and could be a valuable tool for serological surveillance of APPV infection in pigs.

Dyslipidemia is associated with sarcopenia of the elderly: a meta-analysis.

PURPOSE: Sarcopenia is a pathological change characterized by muscle loss in older people. According to the reports, there is controversy on the relationship between dyslipidemia and sarcopenia. Therefore, this meta-analysis aimed to explore the association between sarcopenia and dyslipidemia. METHODS: We searched the Cochrane Library, Web of Science, PubMed, China National Knowledge Infrastructure (CNKI), Wan Fang, China Science and Technology Journal Database (VIP Database) for case‒control studies to extract data on the odds ratio (OR) between sarcopenia and dyslipidemia and the MD(mean difference) of TC, LDL-C, HDL-C, TG, and TG/HDL-C between sarcopenia and nonsarcopenia. The JBI(Joanna Briggs) guidelines were used to evaluate the quality. Excel 2021, Review Manager 5.3 and Stata 16.0 were used for the statistical analysis. RESULTS: Twenty studies were included in the meta-analysis, 19 of which were evaluated as good quality. The overall OR of the relationship between sarcopenia and dyslipidemia was 1.47, and the MD values of TC, LDL-C, HDL-C, TG, and TG/HDL-C were 1.10, 1.95, 1.27, 30.13, and 0.16 respectively. In female, compared with the non-sarcopnia, the MD of TC, LDL-C, HDL-C, TG of sarcopenia were - 1.67,2.21,1.02,-3.18 respectively. In male, the MD of TC, LDL-C, HDL-C, TG between sarcopenia and non-sarcopenia were - 0.51, 1.41, 5.77, -0.67. The OR between sarcopenia and dyslipidemia of the non-China region was 4.38, and it was 0.9 in China. In the group(> 60), MD of TC between sarcopenia and non-sarcopenia was 2.63, while it was 1.54 in the group(20-60). CONCLUSION: Dyslipidemia was associated with sarcopenia in the elderly, which was affected by sex, region and age.

A systematic review of the relationship between normal range of serum thyroid-stimulating hormone and bone mineral density in the postmenopausal women.

OBJECTIVE: The aim of this study was adopts meta-analysis in evaluating the correlation between TSH and BMD, as well as osteoporosis in the postmenopausal women with normal thyroid function. METHODS: Six databases were searched for articles concerning correlation between TSH and BMD in postmenopausal women. The retrieval time was set from the date of database establishment to November 30, 2020. Revman5.3 and Stata12.0 software were used for meta-analysis. RESULTS: A total of 19 articles were incorporated. The Summary Fisher' Z of the correlation between TSH and BMD was 0.16, 95% CI (0.00, 0.32), and the correlation coefficient of Summary Fisher' Z conversion was 0.158. Study on the relationship between TSH and osteoporosis based on OR demonstrated that the combined OR was 1.76, 95% CI (1.27, 2.45), P < 0.05. The subgroup analyzing results displayed that the risk of osteoporosis of the subjects from community with low TSH was 1.89, 95% CI (1.43, 2.49). The risk of osteoporosis for subjects with low TSH and from hospitals was 1.36, 95% CI (0.46, 3.99); 1.84 for subjects with low TSH and anti-osteoporosis drugs, 95% CI (1.05, 3.22); and 1.74 for those with low TSH but not taking anti-osteoporosis drugs, 95% CI (1.08, 2.82). The dose-response relationship showed that the risk of osteoporosis tended to decrease when TSH was more than 2.5mIu/L. CONCLUSION: The serum TSH is positively related with BMD in postmenopausal women, and high TSH (> 2.5 mIu/L) within the normal range is possibly helpful to decrease the risk of osteoporosis in postmenopausal women.

Medical Consultations Option and Influencing Factors for SARS-CoV-2 Infected Individuals - Beijing Municipality, China, December 2022.

What is already known about this topic?: In December 2022, China revised its epidemic prevention and control strategy, leading to an increase in coronavirus disease 2019 (COVID-19) cases and a peak in medical consultations. Government departments implemented relevant policies to coordinate and allocate medical resources throughout China. However, there is a scarcity of research on the status of medical consultations and the factors influencing them. What is added by this report?: In the study population, over 80% of individuals with COVID-19 chose not to pursue medical care, while more than 70% of patients who sought treatment opted for primary healthcare facilities. The decision to consult medical professionals was influenced by various factors, such as age, education level, employment status, urban-rural distribution, and the presence of symptoms following COVID-19 infection. What are the implications for public health practice?: The implementation of tiered diagnostic and treatment approaches, aligned with guidelines issued by governing bodies, is essential for mitigating the strain on medical resources. Primary healthcare institutions serve as "gatekeepers" for public health and should be further expanded in the future.

A community-based cross-sectional study of anxiety and metabolic syndrome.

BACKGROUND: The prevalence of anxiety and other psychological disorders has increased during the COVID-19 pandemic, especially among the elderly. Anxiety and metabolic syndrome (MetS) may aggravate each other. This study further clarified the correlation between the two. METHODS: Adopting a convenience sampling method, this study investigated 162 elderly people over 65 years of age in Fangzhuang Community, Beijing. All participants provided baseline data on sex, age, lifestyle, and health status. The Hamilton Anxiety Scale (HAMA) was used to assess anxiety. Blood samples, abdominal circumference, and blood pressure were used to diagnose MetS. The elderly were divided into MetS and control groups according to the diagnosis of MetS. Differences in anxiety between the two groups were analysed and further stratified by age and gender. Multivariate logistic regression analysis was used to analyse the possible risk factors for MetS. RESULTS: Compared with the control group, anxiety scores of the MetS group were statistically higher (Z = 4.78, P < 0.001). There was a significant correlation between anxiety levels and MetS (r = 0.353, P < 0.001). Multivariate logistic regression revealed that anxiety (possible anxiety vs no anxiety: odds ratio [OR] = 2.982, 95% confidence interval [CI] 1.295-6.969; definite anxiety vs no anxiety: OR = 14.573, 95%CI 3.675-57.788; P < 0.001) and BMI (OR = 1.504, 95% CI 1.275-1.774; P < 0.001) were possible risk factors for MetS. CONCLUSION: The elderly with MetS had higher anxiety scores. Anxiety may be a potential risk factor for MetS, which provides a new perspective on anxiety and MetS.

Identification of Three Campylobacter Lysins and Enhancement of Their Anti-Escherichia coli Efficacy Using Colicin-Based Translocation and Receptor-Binding Domain Fusion.

The emergence of multidrug-resistant Escherichia coli, which poses a major threat to public health, has motivated the development of numerous alternative antimicrobials. Lysins are bacteriophage- and bacterium-derived peptidoglycan hydrolases that represent a new antibiotic treatment targeting bacterial cell walls. However, the bactericidal effect of native lysins on Gram-negative bacteria is restricted by the presence of an outer membrane. Here, we first evaluated the antibacterial activity of three Campylobacter-derived lysins (Clysins) against E. coli. To improve their transmembrane ability and antibacterial activities, six engineered Clysins were constructed by fusing with the translocation and receptor-binding (TRB) domains from two types of colicins (colicin A [TRBA] and colicin K [TRBK]), and their biological activities were determined. Notably, engineered lysin TRBK-Cly02 exhibited the highest bactericidal activity against the E. coli BL21 strain, with a reduction of 6.22 ± 0.34 log units of cells at a concentration of 60.1 µg/mL, and formed an observable inhibition zone even at a dose of 6.01 µg. Moreover, TRBK-Cly02 killed E. coli dose dependently and exhibited the strongest bactericidal activity at pH 6. It also exhibited potential bioactivity against multidrug-resistant E. coli clinical isolates. In summary, this study identified three lysins from Campylobacter strains against E. coli, and the enhancement of their antibacterial activities by TRB domains fusion may allow them to be developed as potential alternatives to antibiotics. IMPORTANCE Three lysins from Campylobacter, namely, Clysins, were investigated, and their antibacterial activities against E. coli were determined for the first time. To overcome the restriction of the outer membrane of Gram-negative bacteria, we combined the TRB domains of colicins with these Clysins. Moreover, we discovered that the Clysins fused with TRB domains from colicin K (TRBK) killed E. coli more effectively, and this provides a new foundation for the development of novel bioengineered lysins by employing TRBK constructs that target outer membrane receptor/transport systems. One of the designed lysins, TRBK-Cly02, exhibited potent bactericidal efficacy against E. coli strains and may be used for control of multidrug-resistant clinical isolates. The results suggest that TRBK-Cly02 can be considered a potential antibacterial agent against pathogenic E. coli.

Screening efficacy of PhA and MNA-SF in different stages of sarcopenia in the older adults in community.

OBJECTIVE: To compare the screening ability of the phase Angle (PhA) and the Short-Form Mini Nutritional Assessment (MNA-SF) alone and combined detection in the different stages of sarcopenia among the older adults in the community. METHODS: The older adults aged 65 and above were enlisted during community outpatient service and their nutritional status was evaluated by MNA-SF scale. PhA was measured by bioelectrical impedance analysis (BIA). AWGS2019 and EWGSOP2010 were used to define the different stages of sarcopenia. We measured skeletal mass index (SMI) and grip strength with BIA and electronic grip apparatus and measured body function with 6-m pace, SPPB test, and standing test. RESULTS: The AUC of PhA in the screening of possible sarcopenia was 0.640, the sensitivity was 58.49%, the specificity was 66.67%, and the cut-off value was 4.5. The AUC of the combined PhA and MNA-SF for possible sarcopenia was 0.642, the sensitivity was 57.55%, and the specificity was 70.00%. The AUC of MNA-SF for the screening of pre-sarcopenia was 0.805, the sensitivity was 66.67%, the specificity was 85.83%, and the cut-off value was 12. The AUC of the combined PhA and MNA-SF was 0.826, the sensitivity was 75.00%, and the specificity was 85.00%. The AUC of PhA in the screening of sarcopenia (common type) was 0.808, the sensitivity was 82.35%, the specificity was 73.33%, the cut-off value was 4.4. The AUC of the combined PhA and MNA-SF for sarcopenia (common type) was 0.835, the sensitivity was 76.47% and the specificity was 81.67%. The AUC of PhA and for the screening of severe sarcopenia was 0.935, the sensitivity was 93.33%, the specificity was 92.50%, and the cut-off value was 4.1. The AUC of the combined PhA and MNA-SF was 0.943, the sensitivity was 86.67%, and the specificity was 93.33%. CONCLUSION: The screening ability of PhA alone or in combination was higher than that of MNA-SF in the screening of possible sarcopenia. The screening ability of the combined detection was higher than that of PhA alone in the screening of pre-sarcopenia. The combination of PhA and MNA-SF or PhA alone all performed better value in the screening of sarcopenia (common type). Compared to MNA-SF, the PhA performed better in the screening of severe sarcopenia, which provided references for identifying patients with different stages of sarcopenia in the community.

Nanosized Shikonin-Fe(III) Coordination Material for Synergistic Wound Treatment: An Initial Explorative Study.

Shikonin (Shik), a natural pigment, has received growing interest in various biomedical fields due to its anti-inflammatory, antitumor, antimicrobial, and antioxidant ability. However, some inherent characteristics of Shik, such as its virulence, low bioavailability, and poor solubility, have limited its biomedical applicability. Here, we reported a facile synthetic method to produce the Shik-iron (III) nanoparticles (Shik-Fe NPs), which could overcome these limitations of Shik. The synthesized Shik-Fe NPs possessed a uniform size range of 110 ± 10 nm, negative surface charges, good water dispersity, and high safety. Iron distributed uniformly inside Shik-Fe NPs, and iron constituted 20% of total mass in PEGylated Shik-Fe NPs. When interacting with activated macrophages, Shik-Fe NPs significantly reduced the level of cellular inflammatory factors, for example, iNOS, IL-1ß, and TNF-α. Furthermore, the Shik-Fe NPs demonstrated synergistic anti-inflammation and anti-bacterial properties in vivo, since they could release Fe3+ and Shik to eradicate bacteria (Staphylococcus aureus and P. aeruginosa were used as model microbes here) during wound infections and provide full recovery for scald wounds. Collectively, the study established a dual-functional Shik-derived nanoplatform, which could be useful for the treatment of various inflammation-involved diseases.

Salvianolic acid B alleviates neurological injury by upregulating stanniocalcin 1 expression.

Background: Salvianolic acid B (Sal B) is a representative component of phenolic acids derived from the dried root and rhizome of Salvia miltiorrhiza Bge. (Labiatae), which promotes angiogenesis in myocardial infarction and diabetic cardiomyopathy. However, whether Sal B has a neuroprotective function in ischemic stroke by promoting angiogenesis is still unclear. Methods: In the present study, ischemic stroke models were induced in rats by middle cerebral artery occlusion (MCAO), and Sal B (10 or 20 mg/kg/d) was intraperitoneally injected according to a previous study. Neurological deficits were evaluated by the modified Longa five-point scale, modified Bederson scores and cerebral infarction sizes by triphenyltetrazolium chloride (TTC) staining. Apoptotic cells were tested by cleaved-caspase3 immunofluorescence staining and an in situ cell death (TUNEL) detection kit. Human umbilical vein endothelial cells (HUVECs) exposed to hypoxia were used to investigate the effects of Sal B on angiogenesis and tube formation in vitro. Results: Sal B ameliorated the neurological deficits, decreased the cerebral infarction volumes in rats with ischemic stroke, significantly increased the expression of vascular endothelial growth factor receptor 2 (VEGFR2) and VEGFA and promoted angiogenesis both in vivo and in vitro. Furthermore, Sal B increased stanniocalcin 1 (STC1) expression, induced the phosphorylation of protein kinase B (AKT) and mammalian target of rapamycin (mTOR) activity, enhanced cell migration, and activated VEGFR2/VEGFA signaling in endothelial cells. Conclusions: This study showed that Sal B promoted angiogenesis and alleviated neurological apoptosis in rats with ischemic stroke by promoting STC1.

ILF3 is responsible for hyperlipidemia-induced arteriosclerotic calcification by mediating BMP2 and STAT1 transcription.

Calcification is common in atherosclerotic plaque and can induce vulnerability, which further leads to myocardial infarction, plaque rupture and stroke. The mechanisms of atherosclerotic calcification are poorly characterized. Interleukin enhancer binding factor 3 (ILF3) has been identified as a novel factor affecting dyslipidemia and stroke subtypes. However, the precise role of ILF3 in atherosclerotic calcification remains unclear. In this study, we used smooth muscle-conditional ILF3 knockout (ILF3SM-KO) and transgenic mice (ILF3SM-Tg) and macrophage-conditional ILF3 knockout (ILF3M-KO) and transgenic (ILF3M-Tg) mice respectively. Here we showed that ILF3 expression is increased in calcified human aortic vascular smooth muscle cells (HAVSMCs) and calcified atherosclerotic plaque in humans and mice. We then found that hyperlipidemia increases ILF3 expression and exacerbates calcification of VSMCs and macrophages by regulating bone morphogenetic protein 2 (BMP2) and signal transducer and activator of transcription 1 (STAT1) transcription. We further explored the molecular mechanisms of ILF3 in atherosclerotic calcification and revealed that ILF3 acts on the promoter regions of BMP2 and STAT1 and mediates BMP2 upregulation and STAT1 downregulation, which promotes atherosclerotic calcification. Our results demonstrate the effect of ILF3 in atherosclerotic calcification. Inhibition of ILF3 may be a useful therapy for preventing and even reversing atherosclerotic calcification.