[Analysis of the types and functions of CD34+ cells in full-thickness skin defect wounds of normal mice and diabetic mice by single-cell RNA sequencing].

Objective: To analyze the types and functions of CD34+ cells in full-thickness skin defect wounds of normal mice and diabetic mice by single-cell RNA sequencing. Methods: This study was an experimental study. The CD34+ cell lineage tracing mouse was produced, and the visualization of CD34+ cells under the fluorescent condition was realized. Six male CD34+ cell lineage tracing mice aged 7-8 weeks (designated as diabetic group) were intraperitoneally injected with streptozotocin to establish a diabetic model, and full-thickness skin defect wounds were prepared on their backs when they reached 13 weeks old. Another 6 male CD34+ cell lineage tracing mice aged 13 weeks (designated as control group) were also subjected to full-thickness skin defect wounds on their backs. On post-injury day (PID) 4, wound tissue was collected from 3 mice in control group and 2 mice in diabetic group, and digested to prepare single-cell suspensions. CD34+ cells were screened using fluorescence-activated cell sorting, followed by single-cell RNA sequencing. The Seurat 4.0.2 program in the R programming language was utilized for dimensionality reduction, visualization, and cell clustering analysis of CD34+ cell types, and to screen and annotate the marker genes for each CD34+ cell subpopulation. Kyoto encyclopedia of genes and genomes (KEGG) and gene ontology (GO) enrichment analysis was performed to analyze the differentially expressed genes (DEGs) of CD34+ fibroblasts (Fbs), smooth muscle cells (SMCs), keratinocytes (KCs), and chondrocyte-like cells (CLCs) in the wound tissue of two groups of mice for exploring cellular functions. Results: On PID 4, CD34+ cells in the wound tissue of both groups of mice were consisted of 7 cell types, specifically endothelial cells, Fbs, KCs, macrophages, T cells, SMCs, and CLCs. Among these, Fbs were further classified into 5 subpopulations. Compared with those in control group, the proportions of CD34+ endothelial cells, Fbs subpopulation 1, Fbs subpopulation 4, KCs, and CLCs in the wound tissue of mice were increased in diabetic group, while the proportions of CD34+ Fbs subpopulation 2, Fbs subpopulation 3, and SMCs were decreased. The marker genes for annotating CD34+ CLCs, endothelial cells, Fbs subpopulation 1, Fbs subpopulation 2, Fbs subpopulation 3, Fbs subpopulation 4, Fbs subpopulation 5, KCs, macrophages, SMCs, and T cells were respectively metastasis-associated lung adenocarcinoma transcript 1, fatty acid binding protein 4, Gremlin 1, complement component 4B, H19 imprinted maternally expressed transcript, Dickkopf Wnt signaling pathway inhibitor 2, fibromodulin, keratin 5, CD74 molecule, regulator of G protein signaling 5, and inducible T-cell co-stimulator molecule. KEGG and GO enrichment analysis revealed that, compared with those in control group, DEGs with significant differential expression (SDE) in CD34+ Fbs from the wound tissue of mice in diabetic group on PID 4 were significantly enriched in terms related to inflammatory response, extracellular matrix (ECM) organization, regulation of cell proliferation, and aging (with Pvalues all <0.05), DEGs with SDE in CD34+ SMCs were significantly enriched in terms related to cell migration, apoptotic process, positive regulation of transcription, and phagosome (with P values all <0.05), DEGs with SDE in CD34+ KCs were significantly enriched in terms related to mitochondrial function, transcription, and neurodegenerative diseases (with P values all <0.05), and DEGs with SDE in CD34+ CLCs were significantly enriched in terms related to rhythm regulation, ECM, and viral infection (with P values all <0.05). Conclusions: CD34+ cells display high heterogeneity in the healing process of full-thickness skin defect wounds in both normal mice and diabetic mice. The significantly enriched functions of DEGs with SDE in CD34+ cell subpopulations in the wound tissue of the two mouse groups are closely related to the wound healing process.

Fruit Intake and Alzheimer's Disease: Results from Mendelian Randomization.

Alzheimer's disease (AD) is the leading cause of dementia in old age, recognized as a global health priority. To explore causal effects of fresh fruit intake and dried fruit intake on AD liability, this study utilized GWAS from the UK Biobank and FinnGen to conduct Mendelian randomization (MR) analysis, and used inverse variance weighted (IVW), MR-Egger, and weighted median approaches for MR estimates, and visual inspections judged result stability. Results suggested little evidence of a potential causal relationship between fresh fruit intake and AD (OR=0.97, 95%CI=0.50-1.91, P=0.939), while significant, robust causality was indicated between dried fruit intake and AD (OR=4.09, 95%CI=2.07-8.10, P<0.001). Stability evaluations showed no heterogeneity or pleiotropy affecting interpretability and credibility of primary analyses. In conclusion, we strengthened evidence for positive causality from dried fruit intake to AD liability, with causality from fresh fruit intake on AD risk was not demonstrated.

Bevacizumab for recurrent glioblastoma: a systematic review and meta-analysis.

OBJECTIVE: The phenomenon is that few randomized control trials (RCTs) directly compared the effects of bevacizumab with other types of standard treatments for recurrent glioblastoma (GBM). We conducted a systematic review and meta-analysis to assess the efficacy of bevacizumab in recurrent GBM patients. MATERIALS AND METHODS: We searched electronic databases (Medline, Embase, and Web of Science) contrasting the bevacizumab with standard treatments up to May 2021. For the continuous outcomes of median progression-free survival (PFS) and median overall survival (OS), we summarized the mean difference (MD) as the effective index. We used relative risk (RR) to estimate the data with a random-effects model to get the outcomes of objective response rate (ORR), 12-month OS, 6-month PFS, and any mentioned adverse events. RESULTS: A total of 807 patients in 5 RCTs included into our systematic review and meta-analysis. The results showed bevacizumab could provide benefits of the ORR (RR, 2.67; 95% CI: 1.14-6.26, p = 0.02), median PFS (MD, 1.12 months; 95% CI: 0.35-1.90 months, p = 0.005), but not the median OS (MD, -0.19 months; 95% CI: -1.37-0.99 months, p = 0.75). Whereas the rates of the secondary outcomes of interest were similar between the bevacizumab group and control group, including 6 month-PFS (RR, 1.23; 95% CI, 0.82-1.84, p = 0.32) and 12 month-OS (RR, 0.93; 95% CI, 0.79-1.09, p = 0.36). As for adverse events, patients with bevacizumab showed higher rates of grade 3/4 and any grade hypertension compared with those with standard treatments (RR, 3.71; 95% CI: 1.17-11.76, p = 0.03; RR, 2.68; 95% CI: 1.26-5.76, p = 0.01, respectively). CONCLUSIONS: This study provides clear proof of the beneficial effects of bevacizumab treatment in recurrent GBM patients. The only observed adverse event was grade 3/4 or any grade hypertension.

[Clinical characteristics and prognosis of intraductal papillary mucinous neoplasm of the biliary tract].

Objective: To study the clinical features, diagnosis, surgery treatment and prognosis of intraductal papillary mucinous neoplasm of the biliary tract (IPMN-B). Methods: The data of 16 patients with IPMN-B admitted to The First Hospital of Jilin University and Xi'an Third Hospital from January 2014 to January 2018 were retrospectively analyzed. Kaplan-Meier method was used to conduct the survival analysis. These patients included 10 males and 6 females, the median age was 57 years. Results: Clinical manifestations were mainly jaundice (11 cases), upper abdominal pain (12 cases) and hyperpyretic chills (4 cases), combined with bile duct stones (14 cases) and hepatic lobe atrophy (2 cases). The average size of the tumor was (2.6±0.7) cm. All of 16 cases were diagnosed as IPMN-B, including 5 cases of invasive carcinoma (4 cases were perineural invasion) without vascular invasion and 6 cases of non-invasion carcinoma. The pathological type included 8 cases of pancreaticobiliary duct type, 5 cases of gastric type, 2 cases of intestinal type and 1 case of eosinophils type. Laboratory tests showed abnormal liver function (12 cases), increased direct bilirubin (9 cases), increased carbohydrate antigen199 (CA199, 8 cases) and carcinoembryonic antigen (CEA, 4 cases). The enhanced CT detection showed 9 cases of intrabile duct mass and 14 cases of bile duct dilatation. Surgical resection is the main treatment method, including 2 cases of partial resection of liver, 12 cases of bile duct mass resection combined with choledochojejunostomy and 2 cases of pancreatoduodenectomy, all of whom achieved R0 resection. Fifteen patients were followed up and 4 died during the period. The median postoperative progression free survival was 31 months (95% CI: 33-47 months), and the recurrence rate at 1 year, 2-years and 3-years were 6.7%, 40.0% and 73.3%, respectively. The median overall survival was 35 months (95% CI: 23-47 months), and the 1 year, 2-years, and 3-years cumulative survival rates were 100%, 80.0%, and 53.3%, respectively. Conclusions: IPMN-B is a rare tumor of biliary tract system, which is difficult to be diagnosed early. The main treatment is surgical resection, which can achieve a good prognosis.

Different fundus imaging modalities and technical factors in AI screening for diabetic retinopathy: a review.

BACKGROUND: Effective screening is a desirable method for the early detection and successful treatment for diabetic retinopathy, and fundus photography is currently the dominant medium for retinal imaging due to its convenience and accessibility. Manual screening using fundus photographs has however involved considerable costs for patients, clinicians and national health systems, which has limited its application particularly in less-developed countries. The advent of artificial intelligence, and in particular deep learning techniques, has however raised the possibility of widespread automated screening. MAIN TEXT: In this review, we first briefly survey major published advances in retinal analysis using artificial intelligence. We take care to separately describe standard multiple-field fundus photography, and the newer modalities of ultra-wide field photography and smartphone-based photography. Finally, we consider several machine learning concepts that have been particularly relevant to the domain and illustrate their usage with extant works. CONCLUSIONS: In the ophthalmology field, it was demonstrated that deep learning tools for diabetic retinopathy show clinically acceptable diagnostic performance when using colour retinal fundus images. Artificial intelligence models are among the most promising solutions to tackle the burden of diabetic retinopathy management in a comprehensive manner. However, future research is crucial to assess the potential clinical deployment, evaluate the cost-effectiveness of different DL systems in clinical practice and improve clinical acceptance.

Seed-borne endophytic Bacillus velezensis LHSB1 mediate the biocontrol of peanut stem rot caused by Sclerotium rolfsii.

AIMS: This study aimed to obtain an antagonistic endophyte against Sclerotium rolfsii from peanut seeds, evaluate the biocontrol efficacy towards peanut stem rot and explore its antifungal mechanism against S. rolfsii. METHODS AND RESULTS: Thirty-seven endophytic bacteria were isolated from peanut seeds, six of which exhibited stronger antagonistic activities against S. rolfsii (inhibition rate, IR of hyphae growth ≥70%). Strain LHSB1, the strongest antagonistic strain, was identified as Bacillus velezensis. LHSB1 showed 93·8% of radial growth inhibition of S. rolfsii hyphae and exhibited obvious antagonistic activity against another six pathogenic fungi of peanut. Pot experiments showed two different LHSB1 treatments both significantly reduced the disease incidence and severity of stem rot (P < 0·05) compared to the controls, and the biocontrol efficacy reached 62·6-70·8%, significantly higher than that of Carbendazim control (P < 0·05). Further analyses revealed LHSB1 culture filtrate significantly inhibited sclerotia formation and germination, caused the abnormalities and membrane integrity damage of S. rolfsii hyphae, which might be the possible mode of action of LHSB1 against S. rolfsii. Three antifungal lipopeptides bacillomycin A, surfactin A and fengycin A, were detected in LHSB1 culture extracts by UPLC-ESI-MS, which could be responsible for the biocontrol activity of LHSB1 against S. rolfsii. CONCLUSION: Our results suggested that the seed-borne endophytic B. velezensis LHSB1 would be a tremendous potential agent for the biocontrol of peanut stem rot caused by S. rolfsii. SIGNIFICANCE AND IMPACT OF THE STUDY: This comprehensive study provides a candidate endophytic biocontrol strain and reveals its antifungal mechanism against S. rolfsi. To the best of our knowledge, this is the first time that seed-borne endophytic B. velezensis was used as the biocontrol agent to control peanut stem rot.

CDK5 Regulates PD-L1 Expression and Cell Maturation in Dendritic Cells of CRSwNP.

The maturation of dendritic cells is critical for chronic rhinosinusitis with nasal polyps (CRSwNPs), especially eosinophilic chronic rhinosinusitis with nasal polyps (EosCRSwNPs), but the regulation mechanism of dendritic cells (DCs) maturation is still unclear. We identified nasal mucosa of 20 patients with EosCRSwNP, 16 non-EosCRSwNP patients, and inferior turbinate of 14 patients with nasal septum deviation after surgery. The expression of cyclin-dependent kinase 5 (CDK5) and programmed cell death 1 ligand 1 (PD-L1) were detected by immunofluorescent, real-time quantitative PCR, and Western blot in EosCRSwNP. The level of dendritic cell maturation was detected by flow cytometry and immunofluorescence staining after CDK5 expression interference with small interfering RNA (siRNA). The expression of CDK5 and PD-L1 in EosCRSwNP nasal mucosal tissue was significantly higher than that of non-EosCRSwNP and inferior turbinate nasal mucosa tissue, and there was a positive correlation between them. Immunofluorescence staining showed that CDK5 and PD-L1 were co-localized in dendritic cells. Synergistic stimulation of dendritic cells with LPS and TNF-α promotes the maturation of dendritic cells and increases the expression of CDK5 and PD-L1. However, blocking the expression of CDK5 in dendritic cells with siRNAs leads to a blockage of cell maturation. CDK5 can regulate the expression of PD-L1, and its presence is critical for the maturation of dendritic cells. CDK5 may play an important role in the pathogenesis of CRSwNP disease.

Correction to: CDK5 Regulates PD-L1 Expression and Cell Maturation in Dendritic Cells of CRSwNP.

The article CDK5 Regulates PD-L1 Expression and Cell Maturation in Dendritic Cells of CRSwNP, written by C. C. Liu, H. L. Zhang, L. L. Zhi, P. Jin, L. Zhao, T. Li, X. M. Zhou, D. S. Sun, G. H. Cheng, Q. Xin, L. Shi, and M. Xia was originally published electronically on the publisher's internet.

Artificial intelligence using deep learning to screen for referable and vision-threatening diabetic retinopathy in Africa: a clinical validation study.

BACKGROUND: Radical measures are required to identify and reduce blindness due to diabetes to achieve the Sustainable Development Goals by 2030. Therefore, we evaluated the accuracy of an artificial intelligence (AI) model using deep learning in a population-based diabetic retinopathy screening programme in Zambia, a lower-middle-income country. METHODS: We adopted an ensemble AI model consisting of a combination of two convolutional neural networks (an adapted VGGNet architecture and a residual neural network architecture) for classifying retinal colour fundus images. We trained our model on 76 370 retinal fundus images from 13 099 patients with diabetes who had participated in the Singapore Integrated Diabetic Retinopathy Program, between 2010 and 2013, which has been published previously. In this clinical validation study, we included all patients with a diagnosis of diabetes that attended a mobile screening unit in five urban centres in the Copperbelt province of Zambia from Feb 1 to June 31, 2012. In our model, referable diabetic retinopathy was defined as moderate non-proliferative diabetic retinopathy or worse, diabetic macular oedema, and ungradable images. Vision-threatening diabetic retinopathy comprised severe non-proliferative and proliferative diabetic retinopathy. We calculated the area under the curve (AUC), sensitivity, and specificity for referable diabetic retinopathy, and sensitivities of vision-threatening diabetic retinopathy and diabetic macular oedema compared with the grading by retinal specialists. We did a multivariate analysis for systemic risk factors and referable diabetic retinopathy between AI and human graders. FINDINGS: A total of 4504 retinal fundus images from 3093 eyes of 1574 Zambians with diabetes were prospectively recruited. Referable diabetic retinopathy was found in 697 (22·5%) eyes, vision-threatening diabetic retinopathy in 171 (5·5%) eyes, and diabetic macular oedema in 249 (8·1%) eyes. The AUC of the AI system for referable diabetic retinopathy was 0·973 (95% CI 0·969-0·978), with corresponding sensitivity of 92·25% (90·10-94·12) and specificity of 89·04% (87·85-90·28). Vision-threatening diabetic retinopathy sensitivity was 99·42% (99·15-99·68) and diabetic macular oedema sensitivity was 97·19% (96·61-97·77). The AI model and human graders showed similar outcomes in referable diabetic retinopathy prevalence detection and systemic risk factors associations. Both the AI model and human graders identified longer duration of diabetes, higher level of glycated haemoglobin, and increased systolic blood pressure as risk factors associated with referable diabetic retinopathy. INTERPRETATION: An AI system shows clinically acceptable performance in detecting referable diabetic retinopathy, vision-threatening diabetic retinopathy, and diabetic macular oedema in population-based diabetic retinopathy screening. This shows the potential application and adoption of such AI technology in an under-resourced African population to reduce the incidence of preventable blindness, even when the model is trained in a different population. FUNDING: National Medical Research Council Health Service Research Grant, Large Collaborative Grant, Ministry of Health, Singapore; the SingHealth Foundation; and the Tanoto Foundation.

Engineering the porosity and acidity of H-Beta zeolite by dealumination for the production of 2-ethylanthraquinone via 2-(4'-ethylbenzoyl)benzoic acid dehydration.

Environmentally-friendly zeolites have been used commercially to replace concentrated sulfuric acid and oleum in the alkylation reactions and dehydration of alcohols. However, moderate activity, associated with access and diffusion limitations, low intramolecular dehydration selectivity, associated with unsatisfactory acidity, and unknown reusability have hampered their industrial implementation in the dehydration of bulky 2-(4'-ethylbenzoyl)benzoic acid (E-BBA) to 2-ethylanthraquinone (2-EAQ). Herein, we have discovered that after being treated with mild HNO3, nano-sized H-Beta zeolite showed outstanding catalytic activity, selectivity and reusability, compared with a commercial oleum catalyst. A number of techniques, such as XRD, XPS, XRF, 29Si MAS NMR, 27Al MQ MAS NMR, FTIR, NH3-TPD, argon physisorption and HR-TEM, have been employed to decouple the interdependence between acidity, porosity and catalytic performance. It was found that mild HNO3 treatment could clean out the extra-framework aluminium deposits and selectively extract the aluminium species on the outer surface of Beta zeolites, which strengthened the acidity of the Brønsted acid sites (Si(OH)Al) inside the H-Beta micropores, thus increasing the possibility of intramolecular dehydration of E-BBA. Moreover, this mild HNO3 treatment also dredged the network of intercrystalline mesopores, alleviating the diffusion constraints. Therefore, through the dual adjustment of acidity and porosity, dealuminated H-Beta zeolite has a promising future in the green synthesis of 2-EAQ.

Distribution of the FMR1 gene in females by race/ethnicity: women with diminished ovarian reserve versus women with normal fertility (SWAN study).

OBJECTIVE: To study whether reported, but inconsistent, associations between the FMR1 CGG repeat lengths in the intermediate, high normal, or low normal range differentiate women diagnosed with diminished ovarian reserve (DOR) from population controls and whether associations vary by race/ethnic group. DESIGN: Case-control study. SETTING: Academic and private fertility clinics. PATIENT(S): DOR cases (n = 129; 95 Whites, 22 Asian, 12 other) from five U.S. fertility clinics were clinically diagnosed, with regular menses and no fragile X syndrome family history. Normal fertility controls (n = 803; 386 Whites, 219 African-Americans, 102 Japanese, 96 Chinese) from the United States-based SWAN Study had one or more menstrual period in the 3 months pre-enrollment, one or more pregnancy, no history of infertility or hormone therapy, and menopause ≥46 years. Previously, the SWAN Chinese and Japanese groups had similar FMR1 CGG repeat lengths, thus they were combined. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): FMR1 CGG repeat lengths. RESULT(S): Median CGG repeats were nearly identical by case/control group. DOR cases had fewer CGG repeats in the shorter FMR1 allele than controls among Whites, but this was not significant among Asians. White cases had fewer CGG repeats in the shorter allele than Asian cases. No significant differences were found in the high normal/intermediate range between cases and controls or by race/ethnic group within cases in the longer allele. CONCLUSION(S): This study refutes prior reports of an association between DOR and high normal/intermediate repeats and confirms an association between DOR and low normal repeats in Whites.

Spectroscopic and molecular docking study on the interaction between salicylic acid and the induced disease-resistant protein OsAAA1 of rice.

The interaction between salicylic acid (SA) and the induced disease-resistant protein OsAAA1 in rice was studied using spectroscopy and molecular docking. Ultraviolet (UV) absorption spectroscopy demonstrated an interaction between OsAAA1 protein and SA. Spectroscopy showed that this interaction was a dynamic quenching process. Synchronous fluorescence spectroscopy (SFS) further revealed that this interaction caused changes in the microenvironment of tyrosine and tryptophan and that the interaction site was closer to the tryptophan residue. The structural model of protein OsAAA1 was determined by homology modeling method, and the molecular docking simulation diagram of OsAAA1 with SA was obtained. These models, in combination with a Ramachandran plot analysis, showed amino acid residues ranging from position 240 to position 420 as the possible site interacting with SA. Among them, Gly389, Lys257 and Glu425 might be three key amino acids that can form hydrogen bonds with SA.

The effect of low dosage of procaine on lung cancer cell proliferation.

OBJECTIVE: Non-small cell lung cancer (NSCLC) is the most common type of human lung cancer leading cause of cancer death worldwide. However, first-line drugs such as gefitinib and erlotinib showed great drug resistance in the clinical. MATERIALS AND METHODS: The cell proliferation was evaluated by MTT assay; gene expression was detected by qPCR assay. The protein expression was analyzed by Western blotting. RESULTS: Our results showed that in mouse models of lung cancer by A549 or NCI-H1975 xenograft, the local anesthetic drug Procaine (PCA) with 50 mg/kg specifically attenuated tumors compared with the vehicle-treated group. In vitro, PCA suppressed both two human NSCLC cell lines A549 and NCI-H1975 proliferation in a lower dose (at nM grade). The cell proliferation marker PCNA was also downregulated after PCA treatment in vivo. Furthermore, low-dose of PCA could inhibit the mRNA expression of the key NSCLC target EGFR selectively in the A549 cells, however, it was not observed in another cell line NCI-H1975, implying a specific signaling by PCA in the cell type. CONCLUSIONS: Taken together, our data indicate that PCA treatment leads to suppression of tumor growth and proliferation in A549 and NCI-H1975, and there is an EGFR transcription pathway by PCA in A549 cells.

FMR1 CGG Repeats: Reference Levels and Race-Ethnic Variation in Women With Normal Fertility (Study of Women's Health Across the Nation).

FMR1 premutation carriers (55-199 CGG repeats), and potentially women with high normal (35-44) or low normal (<28) CGG repeats, are at risk of premature ovarian aging. The scarcity of population data on CGG repeats <45 CGG, and variation in race-ethnicity, makes it difficult to determine true associations. DNA was analyzed for FMR1 CGG repeat lengths from 803 women (386 caucasians, 219 African Americans, 102 Japanese, and 96 Chinese) from the US-based Study of Women's Health Across the Nation (SWAN). Participants had ≥1 menses in the 3 months before enrollment, ≥1 pregnancy, no history of infertility or hormonal therapy, and menopause ≥46 years. Statistical analyses used Fisher exact tests. Among these women with normal reproductive histories, significant FMR1 repeat length differences were found across race-ethnicity for both the longer (P = .0002) and the shorter (P < .0001) alleles. The trinucleotide length variance was greater for non-Asian than Asian women (P < .0001), despite identical median values. Our data indicate that short allele lengths <25 CGG on one or both alleles are more common in non-Asian than Asian women. We confirm the minor allele in the 35 to 39 CGG range among Asians as reported previously. Only 2 (0.3%) premutation carriers were identified. These data demonstrate that FMR1 distributions do vary by race-ethnicity, even within the "normal" range. This study indicates the need to control for race-ethnicity in FMR1 ovarian aging research and provides race-ethnic population data for females separated by allele.

Gene-gene interactions of IRF5, STAT4, IKZF1 and ETS1 in systemic lupus erythematosus.

Interferon (IFN) activation signaling and T helper 17 (Th17)-cell/B-cell regulation play a critical role in the pathogenesis of systemic lupus erythematosus (SLE). Several studies have provided convincing evidence that polymorphisms in IRF5, STAT4, IKZF1 and ETS1 from these pathways may be involved in SLE by affecting gene expression or epistasis. We analyzed the genetic interaction in known SLE susceptibility loci from the four genes in northern Han Chinese. A total of 946 northern Han Chinese participated in this study (370 unrelated SLE patients and 576 healthy controls). Subjects underwent genotyping for the single-nucleotide polymorphisms (SNPs) rs2004640 in IRF5, rs7574865 in STAT4, rs4917014 in IKZF1 and rs1128334 in ETS1 by use of a TaqMan SNP genotyping assay and direct sequencing. Gene-gene interaction analysis involved direct counting, multifactor dimensionality reduction (MDR) and linear regression analysis. SLE patients and controls differed in allele frequencies of rs7574865, rs1128334 (P < 0.001) and rs4917014 (P < 0.01). Direct counting revealed that the frequency of risk homozygote combinations was higher for SLE patients than controls (P < 0.01). Furthermore, 2-, 3- and 4-way gene-gene epistasis in SLE was confirmed by parametric methods and MDR analysis. Gene expression analysis partially supported the findings. Our study confirmed the association of the IFN pathway or Th17/B-cells and the pathogenesis of SLE, and gene-gene interaction in this pathway may increase the risk of SLE.

An intronic variant associated with systemic lupus erythematosus changes the binding affinity of Yinyang1 to downregulate WDFY4.

Two recent genome-wide association studies of East Asian populations revealed three genetic variants in WDFY4/LRRC18 associated with systemic lupus erythematosus (SLE). To identify the gene contributing to this disease susceptibility, we examined the mRNA expression of WDFY4 and LRRC18 in patients with SLE and healthy controls. WDFY4 was significantly downregulated in SLE patients as compared with controls. We used allelic expression and dual-luciferase assays to identify the functional variant. Transcriptional activity was lower for the rs877819A than -G allele. Electrophoretic mobility shift and supershift assays revealed that the transcription factor Yinyang1 (YY1) binds to rs877819, with lower affinity to the A allele, which explained the reduced transcriptional activity. This effect was further confirmed by YY1 small interfering RNA knockdown, overexpression and chromatin immunoprecipitation experiments. rs877819 in WDFY4 might be the functional site associated with SLE by reduced binding of YY1 and downregulating WDFY4 expression.

Study on the interaction between methyl jasmonate and the coiled-coil domain of rice blast resistance protein Pi36 by spectroscopic methods.

Interaction between the coiled-coil domain of rice blast resistance protein Pi36 and methyl-jasmonate (MeJA) was studied by fluorescence and UV-vis spectroscopic techniques. The quenching mechanism of fluorescence of MeJA by this domain was discussed to be a static quenching procedure. Fluorescence quenching was explored to measure the number of binding sites n and apparent binding constants K. The thermodynamics parameters ΔH, ΔG, ΔS were also calculated. The results indicate the binding reaction was not entropy-driven but enthalpy-driven, and hydrophobic binding played major role in the interaction. The binding sites of MeJA with the coiled-coil structural domain of rice blast resistance protein Pi36 were found to approach the microenvironment of both Tyr and Trp by the synchronous fluorescence spectrometry. The distance r between donor (the coiled-coil domain of rice blast resistance protein Pi36) and acceptor (MeJA) was obtained according to Förster theory of non-radioactive energy transfer.

DNA demethylation in PD-1 gene promoter induced by 5-azacytidine activates PD-1 expression on Molt-4 cells.

The expression of the programmed death 1 (PD-1) gene is an indicator of exhausted T-cells with decreased activation and function. It remains unknown, however, whether the methylation status of the PD-1 gene promoter is associated with PD-1 expression level. This study shows the changes of PD-1 expression levels and the demethylation status of the PD-1 promoter region in Molt-4 cells under different concentrations of 5-azacytidine (5-Zac). The result demonstrated that DNA demethylation at PD-1 promoter may contribute to PD-1 overexpression.

The use of micropatterning to control smooth muscle myosin heavy chain expression and limit the response to transforming growth factor ß1 in vascular smooth muscle cells.

In the healthy artery, contractile vascular smooth muscle cells (VSMCs) have an elongated shape and are highly aligned but transition to a synthetic phenotype in culture, while additionally becoming well spread and randomly organized. Thus, controlling VSMC phenotype is a challenge in tissue engineering. In this study, we investigated the effects of micropatterning on contractile protein expression in VSMCs at low and high passage and in the presence of transforming growth factor beta 1 (TGFß1). Micropatterning led to significantly decreased cell area, increased elongation, and increased alignment compared to non-patterned VSMCs independent of passage number. In the presence of serum, micropatterning led to increased smooth muscle myosin heavy chain (SM-MHC) and α-actin expression in low passage VSMCs, but had no effect on high passage VSMCs. Micropatterning was as effective as TGFß1 in up-regulating SM-MHC at low passage; however, micropatterning limited VSMC response to TGFß1 at both low and high passage. Investigation of TGFß receptor 1 revealed higher expression in non-patterned VSMCs compared to patterned at high passage. Our studies demonstrate that micropatterning is an important regulator of SM-MHC expression in contractile VSMCs and that it may provide a mechanism for phenotype stabilization in the presence of growth factors.