Lipidomic and transcriptomic profiles of glycerophospholipid metabolism during Hemerocallis citrina Baroni flowering.

BACKGROUND: Hemerocallis citrina Baroni (daylily) is a horticultural ornamental plant and vegetable with various applications as a raw material in traditional Chinese medicine and as a flavouring agent. Daylily contains many functional substances and is rich in lecithin, which is mostly composed of glycerophospholipids. To study the comprehensive dynamic changes in glycerophospholipid during daylily flowering and the underlying signalling mechanisms, we performed comprehensive, time-resolved lipidomic and transcriptomic analyses of 'Datong Huanghua 6' daylily. RESULTS: Labelling with PKH67 fluorescent antibodies clearly and effectively helped visualise lipid changes in daylily, while relative conductivity and malonaldehyde content detection revealed that the early stages of flowering were controllable processes; however, differences became non-significant after 18 h, indicating cellular damage. In addition, phospholipase D (PLD) and lipoxygenase (LOX) activities increased throughout the flowering process, suggesting that lipid hydrolysis and oxidation had intensified. Lipidomics identified 558 lipids that changed during flowering, with the most different lipids found 12 h before and 12 h after flowering. Transcriptome analysis identified 13 key functional genes and enzymes in the glycerophospholipid metabolic pathway. The two-way orthogonal partial least squares analysis showed that diacylglycerol diphosphate phosphatase correlated strongly and positively with phosphatidic acid (PA)(22:0/18:2), PA(34:2), PA(34:4), and diacylglycerol(18:2/21:0) but negatively with phospholipase C. In addition, ethanolamine phosphotransferase gene and phospholipid-N-methyltransferase gene correlated positively with phosphatidylethanolamine (PE)(16:0/18:2), PE(16:0/18:3), PE(33:2), and lysophosphatidylcholine (16:0) but negatively with PE(34:1). CONCLUSIONS: Overall, this study elucidated changes in the glycerophospholipid metabolism pathway during the daylily flowering process, as well as characteristic genes, thus providing a basis for future studies of glycerophospholipids and signal transduction in daylilies.

Phellopterin attenuates ovarian cancer proliferation and chemoresistance by inhibiting the PU.1/CLEC5A/PI3K-AKT feedback loop.

Ovarian cancer is known as the second leading cause of gynecologic cancer-associated deaths in women worldwide. Developing new and effective compounds to alleviate chemoresistance is an urgent priority in ovarian cancer. Here, we aimed to reveal the biological function and underlying mechanisms of phellopterin, a naturally sourced ingredient of Angelica dahurica, in ovarian cancer progression as well as evaluate the therapeutic potential of phellopterin in ovarian cancer patients. In this investigation, we found that phellopterin mitigated DNA replication and induced cell cycle arrest, apoptosis, and DNA damage, attenuating cell proliferation and chemoresistance of ovarian cancer. Interestingly, bioinformatics analyses of data from our RNA sequencing and The Cancer Genome Atlas ovarian cancer dataset suggested that phellopterin presented anti-cancer activities in ovarian cancer cells by modulating signals affecting ovarian cancer progression and identified phellopterin as a potential compound in improving ovarian cancer patients' prognosis. In addition, the C-Type Lectin Domain Containing 5A (CLEC5A) was demonstrated as a downstream effector of phellopterin and involved in a positive PU.1/CLEC5A/PI3K-AKT feedback loop. Interestingly, phellopterin might inactivate the positive feedback circuit to suppress ovarian cancer progression. Collectively, our investigation revealed that phellopterin mitigated ovarian cancer proliferation and chemoresistance through suppressing the PU.1/CLEC5A/PI3K-AKT feedback loop, and predicted phellopterin as a new and effective cytotoxic drug and CLEC5A as a potential target for the treatment of ovarian cancer.

[Screening and confirmation of pesticide residues in substitutional tea such as orange peel, lotus leaf, pueraria lobata and Pangdahai].

OBJECTIVE: The screening method was established for pesticide residues in substitutional teas such as orange peel, lotus leaf, pueraria lobata and Pangdahai using data base. The method for analyzing of 26 pesticide residues was confirmed in four substitutional teas with gas chromatography-tandem mass spectrometry(GC-MS/MS) and dispersive solid-phase extraction(d-SPE) for sample preparation. METHODS: The 26 types of pesticides were selected as target compounds, including screened out, commonly used in substitutional tea planting and forbidden in agriculture planting. The samples extracted with acetonitrile, and purified by adsorbent(e. g PSA, GCB, C_(18)), the purification solutions were separated on DB-5 MS UI column((15 m+PUU+15 m)×0. 25 mm×0. 25 µm) with programmed temperature and determined by GC-MS/MS in multi-reaction monitoring(MRM) model. The external standard method was applied to quantify the pesticides. RESULTS: The method showed a good linearity(r≥0. 995)in concentration range(2-100 µg/L) for 26 kinds of pesticides. The limits of quantification(LOQ) were 3-40 µg/kg. The average recoveries range were 71. 9%-114. 4% in three add levels of 10, 50, 200 µg/kg, and relative standard deviations of 1. 0%-16. 0%(n=6). In the four matrixes, the pesticides were detected concentrating on orange peel, and there were kinds of residues. CONCLUSION: This method is simple, fast, sensitive, selective and can satisfy the request of pesticide screening and simultaneous analysis of multiple pesticide residues in orange peel, lotus leaf, pueraria lobata and Pangdahai.

[Determination of pesticide residues in substitutional tea by dispersive solid-phase extraction and gas chromatography-triple quadrupole mass spectrometry].

OBJECTIVE: The method was established for determination 14 kinds of psticide residues in substitutional tea using gas chromatography-triple quadrupole mass spectrometry( GC-MS/MS) for instrument method and dispersive solid-phase extraction( dSPE) for sample preparation. METHODS: The samples extracted with acetonitrile, and purified by N-propyl ethlene diamine( PSA), the extracts were separated on DB-5MS column( 30 m × 0. 25 mm, 0. 25 µm) with programmed temperature and determined by GCMS/MS in multi-reaction monitoring( MRM) model. The external standard method was applied to quantify the pesticides. RESULTS: The method showed a good linearity( r ≥0. 999) in certain ranges for 14 kinds of pesticides. The limits of quantification( LOQ)were 4-40 µg/kg. The average recoveries range were 65. 6%-116. 6% in three add levels of 20, 40 and 200 µg/kg, and relative standard deviations( RSD S) of 1. 5%-15. 7%( n = 6). CONCLUSION: This method is simple, fast, sensitive, selective and can satisfy the request of simultaneous analysis of multiple pesticide residues in substitutional tea.

Intranasal immunization with a peptide conjugated to Salmonella flagellin induces both systemic and mucosal peptide-specific antibody responses in mice.

In this study, the mucosal adjuvant activity of Salmonella flagellin as a carrier in a conjugate of EXP153-rFliC was investigated. EXP153-rFliC was made by conjugation of a synthetic B-cell epitope peptide derived from Plasmodium falciparum exported protein-1(EXP153) to recombinant phase 1 flagellin of Salmonella enterica serovar Typhimurium expressed in Escherichia coli (rFliC), and used to immunize BALB/c mice via intranasal instillation. It was found that robust EXP153-specific serum IgG antibodies were induced without additional adjuvant. EXP153-specific sIgA antibodies were also induced, these being detected in bronchoalveolar, nasal, vaginal and intestinal washes. These observations demonstrate that Salmonella flagellin as a carrier is an effective mucosal adjuvant in that its conjugated peptide induces antibody responses.

[Potential role of CRELD1 gene in the pathogenesis of atrioventricular septal defect].

OBJECTIVE: To screen potential mutation of the CRELD1 gene in congenital atrioventricular septal defect (AVSD) and explore its functional implications. METHODS: Fragments encompassing the 11 coding exons of CRELD1 gene, including at least 50 bp of flanking intronic regions, were amplified with PCR and subjected to DNA sequencing. Results of sequencing were compared with predicted sequence from the GenBank database. Eukaryotic expression vector pcDNA3.1CRELD1 containing the mutational sequence was constructed. Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction (FQ RT-PCR) was applied to examine the expression of CRELD1, Tenascin C and Aggrecan. RESULTS: C857G was identified in a girl with an isolated partial AVSD. The mutation has resulted in a substitution of Alanine for Proline at amino acid 286 in the first cbEGF domain. Western blotting and FQ RT-PCR confirmed that the P286R missense mutation has been a gain-of-function mutation. Compared with the unloaded control, the Aggrecan mRNA expression was downregulated for both wild-type and mutant type samples (t=140.27 vs. 26.36, P < 0.01). The downregulation was more significant in mutant type (t=25.69, P=0.002). There was no significant difference of the Tenascin C expression between wild-type and the unload control (t=1.167, P> 0.05), whilst the Tenascin C expression was up-regulated in mutant type (t=6.66, P=0.022). CONCLUSION: Mutation of the CRELD1 gene may increase the risk for AVSD rather than being directly causative. The P286R mutation of CRELD1 can downregulate the expression of Aggrecan and upregulates the expression of Tenascin C protein, both of which are crucial to extracellular matrix in the formation of the atrioventricular septum. The P286R mutation of CRELD1 may be correlated to the occurrence of AVSD.

Comparative analysis of chloroplast genome and evolutionary history of Hemerocallis.

Members of the genus Hemerocallis have significant value as ornamental, edible, and medicinal plants, particularly in China, where they have been utilized for thousands of years as both a vegetable and Traditional Chinese Medicine. Hemerocallis species exhibit strict control over flowering time, with individuals flowering either diurnally or nocturnally. However, our understanding of the evolutionary history of this genus, especially concerning important horticultural traits, remains limited. In this study, sequencing and assembly efforts were conducted on 73 samples within the Hemerocallis genus. All accessions were classified into two distinct groups based on their diurnal (daylilies) or nocturnal (nightlilies) flowering habits. Comparative analysis of the chloroplast genomes from these two groups identified fifteen variant hotspot regions, including fourteen SNPs and one deletion, which hold promise for the development of molecular markers for interspecific identification. Phylogenetic trees, generated through both maximum-likelihood and Bayesian inference methods using 76 shared protein-coding sequences, revealed that diurnal flowering evolved prior to nocturnal flowering. The divergence between the two groups is estimated to have occurred approximately 0.82 MYA (95% CI: 0.35-1.45 MYA). The ancestral state of Hemerocallis is hypothesized to have featured diurnal flowering with orange yellow petals. This study marks the first reconstruction of the evolutionary history and ancestral state of the genus Hemerocallis. The findings contribute significantly to our understanding of the adaptation and speciation history within the genus.

Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-gamma and PTEN.

Wound healing is essential for maintaining the integrity of multicellular organisms. In every species studied, disruption of an epithelial layer instantaneously generates endogenous electric fields, which have been proposed to be important in wound healing. The identity of signalling pathways that guide both cell migration to electric cues and electric-field-induced wound healing have not been elucidated at a genetic level. Here we show that electric fields, of a strength equal to those detected endogenously, direct cell migration during wound healing as a prime directional cue. Manipulation of endogenous wound electric fields affects wound healing in vivo. Electric stimulation triggers activation of Src and inositol-phospholipid signalling, which polarizes in the direction of cell migration. Notably, genetic disruption of phosphatidylinositol-3-OH kinase-gamma (PI(3)Kgamma) decreases electric-field-induced signalling and abolishes directed movements of healing epithelium in response to electric signals. Deletion of the tumour suppressor phosphatase and tensin homolog (PTEN) enhances signalling and electrotactic responses. These data identify genes essential for electrical-signal-induced wound healing and show that PI(3)Kgamma and PTEN control electrotaxis.

ReLU-FCM trained by quasi-oppositional bare bone imperialist competition algorithm for predicting employment rate.

Fuzzy cognitive maps (FCMs) are a powerful tool for system modeling, which can be used for static and dynamic analysis. However, traditional FCMs are usually learned by gradient-based methods, and the adopted sigmoid nonlinear activation function frequently causes gradient saturation. These two shortcomings set a limit on the modeling accuracy. To overcome those problems, we propose in this paper a new FCM with two improvements. First, the rectified linear unit (ReLu) activation function is adopted to replace the sigmoid function. Second, a newly proposed quasi-oppositional bare bone imperialist competition algorithm (QBBICA) is used to learn the FCM. The improved FCM is used to predict the employment rate of graduates from Liren College, Yanshan University. Experimental results show that the improved FCM is effective in employment rate prediction.

Circular RNA CDK6 suppresses cervical cancer proliferation and metastasis by sponging miR-449a.

Cervical cancer is a malignant tumor that severely threatens female health. Recently, more and more studies indicated that circRNA could function as a tumor activator or suppressor in cervical cell development. Therefore, we aimed to study the effect of circRNA CDK6 (circCDK6) on the development and biological behavior of cervical cancer. We used quantitative real-time PCR (qRT-PCR) to examine the circCDK6 expression level in cervical cancer cell lines. RNA-fluorescence in situ hybridization confirmed the location of circCDK6 and miR-449a in HeLa and CaSki cells, respectively. Then, the biological function of silencing circCDK6 in cellular proliferation, metastasis, and Epithelial-Mesenchymal Transition (EMT)-related process was determined. We also performed RNA Binding Protein Immunoprecipitation (RIP) and Dual-luciferase reporter assay to determine the relationship between the circCDK6 and miR-449a. Finally, the results showed that circCDK6 level remarkably increased in several cervical cancer cells, especially in Hela and CaSki cells. The miR-449a was further confirmed to be a potential target of circCDK6, and its expression increased by silencing circCDK6. The circCDK6 participated in tumorigenesis and cancer progression and might serve as a tumor suppressive factor in cervical cell progression via Epithelial-MesenchymalTransition (EMT) process by regulating miR-449a.

Prevalence of multiple sclerosis in Guangzhou, China: A population-based case-finding prospective study.

BACKGROUND: Multiple sclerosis (MS) is rare in China, and the prevalence previously reported may be biased. Currently, few studies that have investigated the prevalence of MS in China based on the latest diagnostic criteria. METHODS: Through a population-based survey from August 8, 2021 to December 31, 2021, we calculated the prevalence of multiple sclerosis in 18,676,605 residents of Guangzhou, China. MS patients were identified through the health insurance system of the Guangzhou Health Insurance Bureau, and we surveyed 17 large tertiary hospitals using a case-finding approach. All MS patients were diagnosed according to the 2017 McDonald criteria. RESULTS: A total of 143 patients in the resident population of Guangzhou were diagnosed with MS, with a crude prevalence of 0.77 per 100,000 (95% confidence interval (CI): 0.65-0.90), and the prevalence was higher in in females (1.14/100,000) than in males (0.44/100,000). The age-adjusted prevalence was 0.92 per 100,000 (95% CI: 0.77-1.10). The prevalence peaked at the age of 25-29 years (2.86/100,000) for both males and females (1.44/100,000 and 4.42/100,000, respectively). CONCLUSIONS: This is the first study to report the prevalence of MS in Guangzhou, China, according to the criteria. Our study shows that the prevalence of MS in Guangzhou is lower than that in other cities in China.

An improved method of human keratinocyte culture from skin explants: cell expansion is linked to markers of activated progenitor cells.

Human keratinocyte primary cultures are commonly established by tissue dissociation and often rely on feeder cell supports and culture medium that is not defined. Further, contamination by unwanted fibroblasts can be problematic. Here, we developed a skin explant method for growing primary keratinocytes that was rapid, simple, and reliably generated keratinocyte cultures free of fibroblast contamination. The process capitalized on the observation that fibroblasts migrate out of adult skin explants later than epidermal cells, allowing the early harvesting of keratinocytes by trypsinization. When grown subsequently in defined medium in the absence of feeder cells, the explant-derived cells grew rapidly and could be cultured for multiple passages. Immunofluorescence microscopy revealed that a high percentage of cells harvested from the explant outgrowths expressed K15, while very few expressed the differentiation marker K10. Cells that were stained while migrating out from explants strongly expressed markers associated with progenitor cells, including p63, K15 and CD133, and displayed intense K6 expression, indicative of activated keratinocytes in wound-healing epidermis. By replenishing the explants with fresh medium after harvesting, further epidermal outgrowths could be obtained, offering the possibility of greatly increased keratinocyte yields for clinical applications.

Parental attitudes and willingness to donate children's biospecimens for congenital heart disease research: a cross-sectional study in Shanghai, China.

OBJECTIVES: To assess attitudes and willingness of parents of children with congenital heart disease (CHD) regarding donating biospecimens for future CHD research, and to identify factors associated with biospecimen donation. DESIGN: Face-to-face cross-sectional survey data were analysed using logistic regression. SETTING: Cardiothoracic Surgery Inpatient Department, Shanghai Children's Medical Centre. PARTICIPANTS: Parents of children attending the cardiothoracic surgery inpatient department at Shanghai Children's Medical Center, 1 March-31 December 2016. PRIMARY AND SECONDARY OUTCOME MEASURES: Willingness and motivation regarding donating children's biospecimens, and ethical and legal considerations concerning children's future willingness to donate. RESULTS: Of 550 parents, 508 completed the questionnaire (response rate=92.4%). Overall, 69.1% (n=351) were willing to donate their children's biospecimens for medical research. Multivariate analysis indicated higher education level (college/graduate degree: OR 2.435, 95% CI 1.221 to 4.857, p=0.012; high school: OR 1.827, 95% CI 1.190 to 2.804, p=0.006) and children's hospitalisation history (OR 1.581; 95% CI 1.069 to 2.338, p=0.022) were positively associated with willingness to donate. The most common motivation for donation was potential benefit to other children with CHD (81.2%, n=285). The main barriers to donation were physical discomfort to their children (52.3%, n=54) and concerns about personal privacy (47.1%, n=48). Most parents (86.0%, n=302) wanted to be informed of research results using their children's donated biospecimens, and 34.8% (n=177) believed that children aged 10-18 years had the right to consent independently to research participation. CONCLUSIONS: Nearly 70% of the parents in this study were willing to donate their children's biospecimens for future CHD research. Parents' education level and children's hospitalisation history influenced willingness to donate. Most parents wanted to receive the research results related to their children's biospecimens.

Transcriptome profiling of cancer and normal tissues from cervical squamous cancer patients by deep sequencing.

Cervical cancer is the fourth leading cause of cancer mortality in women worldwide. High­risk human papillomavirus infection is a major cause of cervical cancer. A previous study revealed the role of different oncogenes and tumor suppressors in cervical cancer initiation and progression. However, the complicated genetic network regulating cervical cancer remains largely unknown. The present study reported transcriptome sequencing analysis of three cervical squamous cell cancer tissues and paired normal cervical tissues. Transcriptomic analysis revealed that 2,519 genes were differently expressed between cervical cancer tissues and their corresponding normal tissues. Among these, 236 differentially expressed genes (DEGs) were statistically significant, including many DEGs that were novel in cervical cancer, including gastrulation brain homeobox 2,5­hydroxytryptamine receptor 1D and endothelin 3. These 236 significant DEGs were highly enriched in 28 functional gene ontology categories. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis suggested involvement of these DEGs in multiple pathways. The present study provides a transcriptome landscape of cervical cancer in Chinese patients and an improved understanding of the genetic regulatory network in cervical cancer tumorigenesis.

Ameliorative effects of Qingfei Tongluo formula on experimental mycoplasmal pneumonia in mice.

Mycoplasma pneumoniae pneumonia (MPP) is a common disease in children. Qingfei Tongluo formula (QTF) has been used for the treatment of MPP clinically, but the chemical constituents and mechanism involved remain unclear. This study aimed to analyze the main chemical constituents and to explore the possible mechanism of action associated with QTF treatment of MPP. Liquid chromatography-mass spectrometry was employed to identify the compounds contained in the QTF extract. A BALB/c mouse model of MP infection was established. After treatment with QTF (0.85 and 1.70 g/kg) for 3 days, hematoxylin and eosin staining was performed in lung tissues for histological examination. Inflammatory cytokines were detected by ELISA. Western blot analysis was used for detecting phosphorylated proteins involved in MAPK and nuclear factor (NF)-κB signaling pathways. In the mouse model, a large amount of pulmonary interstitial infiltration of lymphocytes and plasmacytes were seen as well as bronchus and vasodilation congestion. Following QTF treatment, inflammation was alleviated significantly compared with the model group. Inflammatory cytokines [interleukin (IL)-6, transforming growth factor-ß1, IL-8, IL-1ß and tumor necrosis factor-α] in bronchoalveolar lavage fluid were decreased dramatically. In addition, we found that QTF inhibited activation of phosphorylation of JNK, ERK and NF-κB. In conclusion, QTF alleviates MPP inflammation possibly via inhibitory activation of MAPK/NF-κB pathways, which can act as a new agent for MPP treatment.

Association of coronary artery calcium with bone mineral density in postmenopausal women.

OBJECTIVES: Atherosclerosis and osteoporosis (OP) are common diseases in elderly individuals and may share common pathogenetic mechanisms. The aim of this study was to investigate the association between bone mineral density (BMD) and coronary artery calcium (CAC) in postmenopausal women. METHODS: In this cross-sectional study, 186 postmenopausal women 50-80 years of age were included. BMD of the spine and femoral neck was measured by dual-energy X-ray absorptiometry. The coronary artery calcium score (CACS) was measured by multidetector computed tomography. RESULTS: The study included postmenopausal women aged 65.6±7.3 years, 109 of whom (58.6%) showed CAC. Thirty-three (17.7%) of the patients were found to have OP in the lumbar spine and 83 (44.6%) had osteopenia, whereas in the femoral neck, 26 patients (14.0%) had OP and 87 patients (46.8%) had osteopenia. The mean CACS was 57.6±108.3 in normal status, 89.7±143.5 in OP, and 156.4±256.9 in osteopenia at the spine (P<0.05). The mean CACS was 43.2±89.9 in normal status, 126.9±180.3 in OP, and 198.2±301.2 in osteopenia at the femoral neck (P<0.05). Multivariable logistic regression analysis showed that BMD was an independent marker for an increased risk of developing CAC in postmenopausal women. The multiple regression model showed that T-scores were the independent predictors of CACS. CONCLUSION: BMD identified on images from dual-energy X-ray absorptiometry were strongly related to multidetector computed tomography measures of CAC. This low-cost, minimal radiation technique used widely for OP screening is a promising marker of generalized coronary atherosclerosis.

Intensive Atorvastatin Therapy Attenuates the Inflammatory Responses in Monocytes of Patients with Unstable Angina Undergoing Percutaneous Coronary Intervention via Peroxisome Proliferator-Activated Receptor γ Activation.

Periprocedural myocardial injury is a prognostically important complication of percutaneous coronary intervention (PCI). However, it still remains unclear whether and how intensive atorvastatin therapy attenuates the unfavorable inflammatory responses of monocytes associated with PCI. The aim of the study was to investigate the impact of intensive atorvastatin therapy on inflammatory responses of monocytes in Chinese patients with unstable angina who received PCI in order to explore the potential anti-inflammatory mechanism. Ninety-six patients with unstable angina were randomly assigned to atorvastatin 80 mg (intensive) or atorvastatin 20 mg (conventional) treatment at a 1:1 ratio. Creatine kinase MB (CK-MB), cTnI, hs-CRP, and IL-6 were assessed, and circulating CD14(+) monocytes were simultaneously obtained using CD14 MicroBeads 2 h before and 24 h after PCI. Plasma levels of CK-MB, cTnI, hs-CRP, and IL-6 were higher in the conventional dose group versus those in the intensive dose group following PCI. Furthermore, intensive atorvastatin treatment markedly reduced the expressions and responses of Toll-like receptor 2 (TLR2), TLR4, and CCR2 of CD14(+) monocytes versus the conventional dose group and significantly increased the activated peroxisome-proliferator-activated receptor (PPAR) γ in the CD14(+) monocytes post-PCI. Notably, the changes in responses of TLR2, TLR4, and CCR2 of CD14(+) monocytes between the two groups were all reversed by PPARγ antagonist and augmented by PPARγ agonist. In conclusion, a single high (80 mg) loading dose of atorvastatin reduced the inflammatory response in Chinese patients with unstable angina following PCI. The anti-inflammatory role of intensive atorvastatin was possibly due to attenuation of inflammatory response in monocytes via PPARγ activation.

Salmonella flagellin is a potent carrier-adjuvant for peptide conjugate to induce peptide-specific antibody response in mice.

As an agonist to innate immune system, Salmonella flagellin has been proven to be a potent adjuvant either admixed or genetically fused with antigens and applied to a variety of vaccines against infectious diseases. However, relatively little is known about its carrier-adjuvant effect for conjugate vaccine. Conjugation is an effective approach often used to make haptens such as some peptides and polysaccharides immunogenic and in some cases used to make poor immunogens more immunogenic. In the current study, Salmonella flagellin was tested for its carrier-adjuvant effect in a peptide conjugation. The recombinant Salmonella flagellin (rFliC) purified from Escherichia coli was firstly modified by maleimide groups, then coupled with a synthetic peptide (EXP153:CDNNLVSGP) that is a B-cell epitope derived from Plasmodium falciparum exported protein-1 to generate the conjugate of EXP153-rFliC. Bioactivity assay showed that both chemical modification and conjugation did not apparently impair the TLR5-ligand activity of rFliC. EXP153-rFliC was used to immunize BALB/c mice via subcutaneous route, and the sera obtained from immunized mice were examined by ELISA and IFA. While no detectable antibody responses were induced by the peptide admixed with rFliC, the robust peptide-specific antibody responses were observed in mice immunized with the peptide conjugated to rFliC in the absence of any additional adjuvant. The immune sera induced by the conjugate recognized the native protein of malaria parasite. The data obtained from this study demonstrate the carrier-adjuvant activity of Salmonella flagellin in peptide conjugate immunization and indicate its promising application for conjugate vaccine research and development.

Preliminary findings on the reliability and validity of the Cantonese Birmingham Cognitive Screen in patients with acute ischemic stroke.

BACKGROUND: There are no currently effective cognitive assessment tools for patients who have suffered stroke in the People's Republic of China. The Birmingham Cognitive Screen (BCoS) has been shown to be a promising tool for revealing patients' poststroke cognitive deficits in specific domains, which facilitates more individually designed rehabilitation in the long run. Hence we examined the reliability and validity of a Cantonese version BCoS in patients with acute ischemic stroke, in Guangzhou. METHOD: A total of 98 patients with acute ischemic stroke were assessed with the Cantonese version of the BCoS, and an additional 133 healthy individuals were recruited as controls. Apart from the BCoS, the patients also completed a number of external cognitive tests, including the Montreal Cognitive Assessment Test (MoCA), Mini Mental State Examination (MMSE), Albert's cancellation test, the Rey-Osterrieth Complex Figure Test, and six gesture matching tasks. Cutoff scores for failing each subtest, ie, deficits, were computed based on the performance of the controls. The validity and reliability of the Cantonese BCoS were examined, as well as interrater and test-retest reliability. We also compared the proportions of cases being classified as deficits in controlled attention, memory, character writing, and praxis, between patients with and without spoken language impairment. RESULTS: Analyses showed high test-retest reliability and agreement across independent raters on the qualitative aspects of measurement. Significant correlations were observed between the subtests of the Cantonese BCoS and the other external cognitive tests, providing evidence for convergent validity of the Cantonese BCoS. The screen was also able to generate measures of cognitive functions that were relatively uncontaminated by the presence of aphasia. CONCLUSION: This study suggests good reliability and validity of the Cantonese version of the BCoS. The Cantonese BCoS is a very promising tool for the detection of cognitive problems in Cantonese speakers.