An integrated method for IgG N-glycans enrichment and analysis: Understanding the role of IgG glycosylation in diabetic foot ulcer.

Diabetic foot ulcer (DFU) is the most common and serious complication of diabetes, and its incidence, disability, and mortality rates are increasing worldwide. The pathogenesis of DFU is associated with dysregulated inflammation mediated by abnormal immunoglobulin G (IgG) glycosylation. In this study, we developed a comprehensive method for IgG N-linked glycosylation in the serum of DFU patients. Through analysis, we identified 31 IgG1 glycans, 32 IgG2 glycans, and 30 IgG4 glycans in the DFU serum. Furthermore, 13 IgG1 glycans, 12 IgG2 glycans, and 5 IgG4 glycans in the DFU groups were found to be significantly different from those of the control groups (p < 0.05). Of these, compared with the control group, one glycan was unique to DFU patients, and seven glycans were not detected in the DFU group. In terms of glycan characteristics, we observed a substantial decrease in galactosylation, sialylation and bisecting GlcNAcylation, and a significant increase in agalactosylation. Abnormal IgG N-glycosylation modifications were significantly associated with the chronic inflammation that is characteristic of DFU. Further, this is the first comprehensive analysis of subclass-specific IgG N-glycosylation in DFU patients, which not only fills the gap of DFU in terms of the pathological mechanisms related to IgG glycosylation but also may provide valuable clues for the immunotherapeutic pathway of DFU.

Hydroxysafflor Yellow A Promotes HaCaT Cell Proliferation and Migration by Regulating HBEGF/EGFR and PI3K/AKT Pathways and Circ_0084443.

OBJECTIVE: To investigate the effect and possible mechanism of hydroxysafflor yellow A (HSYA) on human immortalized keratinocyte cell proliferation and migration. METHODS: HaCaT cells were treated with HSYA. Cell proliferation was detected by the cell counting kit-8 assay, and cell migration was measured using wound healing assay and Transwell migration assay. The mRNA and protein expression levels of heparin-binding epidermal growth factor (EGF)-like growth factor (HBEGF), EGF receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR), and hypoxia-inducible factor-1α (HIF-1α) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Circ_0084443-overexpressing HaCaT cells and empty plasmid HaCaT cells were constructed using the lentiviral stable transfection and treated with HSYA. The expression of circ_0084443 was detected by qRT-PCR. RESULTS: HSYA (800 µmol/L) significantly promoted HaCaT cell proliferation and migration (P<0.05 or P<0.01). It also increased the mRNA and protein expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and increased the phosphorylation levels of PI3K and AKT (P<0.05 or P<0.01). Furthermore, HSYA promoted HaCaT cell proliferation and migration via the HBEGF/EGFR and PI3K/AKT/mTOR signaling pathways (P<0.01). Circ_0084443 attenuated the mRNA expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α (P<0.05). HSYA inhibited the circ_0084443 expression, further antagonized the inhibition of circ_0084443 on HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and promoted the proliferation of circ_0084443-overexpressing HaCaT cells (P<0.05 or P<0.01). However, HSYA could not influence the inhibitory effect of circ_0084443 on HaCaT cell migration (P>0.05). CONCLUSION: HSYA played an accelerative role in HaCaT cell proliferation and migration, which may be attributable to activating HBEGF/EGFR and PI3K/AKT signaling pathways, and had a particular inhibitory effect on the keratinocyte negative regulator circ_0084443.

Triple Ligand Engineered Gold Nanoclusters with Enhanced Fluorescence and Device Compatibility for Efficient Electroluminescence Light-Emitting Diodes.

Gold nanoclusters (Au NCs) are potential emitters for electroluminescent light-emitting diodes (EL-LEDs) but restricted by the limited photoluminescence quantum yield (PLQY) and poor device compatibility. Herein, triple ligand engineered Au NCs enable the fabrication of Au NC-based LEDs with improved EL efficiency. Rigidified triple ligand shells greatly reduce the nonradiative transition and thus increase the PLQY of Au NCs from 2.1 to 73.4%. Most importantly, this strategy significantly improves the compatibility between Au NCs and charge transport materials in EL-LED fabrication. As a result, the EL-LEDs reach a maximum brightness of 1104 cd/m2 and an external quantum efficiency of 5.1%, which is the highest recorded for any reported Au NC-based EL-LEDs.

[Application of copy number variation sequencing in patients with intellectual disability/developmental delay and autistic spectrum disorder].

OBJECTIVE: To assess the value of copy number variation sequencing (CNV-seq) for the diagnosis of children with intellectual disability (ID), developmental delay (DD), and autistic spectrum disorder (ASD). METHODS: Forty patients with ID/DD/ASD referred to Nanshan Maternity and Child Health Care Hospital from September 2018 to January 2022 were enrolled. G-banded karyotyping analysis was carried out for the patients. Genomic DNA was extracted from peripheral blood samples and subjected to CNV-Seq analysis to detect chromosome copy number variations (CNVs) in such patients. ClinVar, DECIPHER, OMIM and other database were searched for data annotation. RESULTS: Among the 40 patients (including 30 males and 10 females), 16, 15 and 6 were diagnosed with ID, DD and ASD, respectively. One patient had combined symptoms of ID and DD, whilst the remaining two had combined ID and ASD. Four patients were found with abnormal karyotypes, including 47,XY,+mar, 46,XY,inv(8)(p11.2q21.2), 46,XX,del(5)(p14) and 46,XX[76]/46,X,dup(X)(p21.1q12). Chromosome polymorphism was also found in two other patients. CNV-seq analysis has detected 32 CNVs in 20 patients (50.0%, 20/40). Pathogenic CNVs were found in 10 patients (25.0%), 15 CNVs of uncertain clinical significance were found in 12 patients (30.0%), and 7 likely benign CNVs were found in 4 patients (10.0%). CONCLUSION: Chromosome CNVs play an important role in the pathogenesis of ID/DD/ASD. CNV-seq can detect chromosomal abnormalities including microdeletions and microduplications, which could provide a powerful tool for revealing the genetic etiology of ID/DD/ASD patients.

Paeoniflorin attenuates the allergic contact dermatitis response via inhibiting the IFN-γ production and the NF-κB/IκBα signaling pathway in T lymphocytes.

Paeoniflorin (PF) has been demonstrated to have an anti-allergic and anti-inflammatory effect in the treatment of allergic contact dermatitis (ACD). However, its clinical application is hampered by the lacking of comprehensive mechanical explanation. This research aimed to study the effect of PF on the proliferation, apoptosis and cytokines secretion as well as the expression of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways of T lymphocytes activation in vitro and in vivo. We found that PF depressed human T lymphocytes activation via inhibition ofinterferon-gamma (IFN-γ) production and NF-κB/IκBα and p38 MAPK signaling pathway in vitro, also PF could attenuate such ACD responses by inhibiting the production of IFN-γ and NF-κB/IκBα pathway in T lymphocytes of ACD mouse model, suggesting that PF might be useful for the treatment of T cell-mediated allergic inflammatory disorders such as ACD. This would make PF a promising T cell-targeted drug candidate for further study because of its immunosuppressive and anti-inflammatory effects.

[Analysis of results of concurrent hearing and deafness genetic screening and follow up of 33 911 newborns].

OBJECTIVE: To analyze the results of concurrent hearing and deafness genetic screening and follow up of newborns. METHODS: In total 33 911 babies born to 5 designated hospitals in Nanshan District of Shenzhen city from October 2017 to December 2019 were included. All subjects underwent concurrent hearing and deafness genetic screening covering 21 variants of 4 genes including GJB2, SLC26A4, GJB3 and Mt12SrRNA. For those with positive results, Sanger sequencing was carried out for confirmation. RESULTS: 93.32% subjects passed the first-round hearing screening, and 87.01% passed the recheck testing. The overall detection rate was 4.18%. The detection rates for GJB2, SLC26A4, GJB3 and Mt12srRNA variants were 1.98%, 1.58%, 0.37% and 0.25%, respectively. 126 and 84 subjects were found with high risk for delayed-onset and drug-induced hearing loss, respectively. In addition, 4 and 5 subjects were found to harbor homozygous/compound heterozygous variants of the GJB2 and SLC26A4 genes, respectively. Concurrent screening showed that subjects (with heterozygous variants) who did not passed the two round hearing test were as follows: GJB2 with 6.75% in the first round and 2.61% in the second round testing, SLC26A4 (3.3%/1.2%), GJB3 (0.72%/0.14%) and 12SrRNA (0.36%/Nil), respectively. Moreover, the No-pass rate in the subjects with homozygous or compound variants in single gene, heterozygous variant in single gene, heterozygous variant in multiple genes, and homozygous variant in GJB3 gene were significantly higher than the subjects with negative results of genetic screening. CONCLUSION: Concurrent newborn genetic screening can enhance the effectiveness of hearing screening and enable earlier identification and intervention for children with hearing impairment. Follow-up can improve the diagnostic rate for children who are positive for the concurrent screening. Nevertheless, genetic and hearing screening cannot replace the diagnostic testing. It is necessary to conduct comprehensive analysis for the results of genetic and hearing screening and radiological examinations. Sanger sequencing and next-generation sequencing are critical for ascertain the diagnosis.

Circular RNA: A novel potential biomarker for skin diseases.

Circular RNA (circRNA) has been classified as noncoding RNA with a covalent closed continuous loop, the 3'and 5' ends of which are normally joined together to increase its own stability. More recently, circRNA has been shown to encode proteins and may be involved in the regulation of gene transcription. This provides more evidence for the involvement of circRNA in disease progression. Accumulating investigations have found that the expression of many circRNAs is abnormal in plenty of skin diseases such as malignant melanoma, psoriasis, and abnormal wound healing. Herein, in addition to the summary of recent studies on the nuclear export, N6-methyladenosine (m6A) modification, degradation, and other biogenesis and properties of circRNA, we highlight the importance of circRNAin skin diseases. Although their exact roles and mechanisms in most skin disease remain preliminary, circRNAs have potential applications as diagnostic biomarkers and novel therapeutic targets for skin diseases due to its structural and functional properties (stability, specificity and sensitivity), which is worthy of deeper exploration and greater research efforts.