Analysis of pathogenic factors of Candida albicans and the effect of vaginal immunization on recurrent vulvovaginal candidiasis in mice.

BACKGROUND: The role of fungal pathogenic factors and the immune response of the vaginal epithelium in vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC) are still unclear. Our study wants to clarify whether there are differences in pathogenic factors between VVC and RVVC strains, confirm the roles of pathogenic factors in the pathogenesis of RVVC, and analyze the influence of pathogenic factors on vaginal host immunity. METHODS: VVC- and RVVC-causing Candida albicans strains were genotyped with 25S rDNA. Drug susceptibility assays using a modified alamarBlue broth microdilution method were carried out. Milk culture medium and egg yolk culture medium were used to measure the secreted aspartate protease (Sap) and phospholipase (Plb) activity of the samples. We used C. albicans with different Sap activity levels to induce RVVC in mice and measured interleukin 4 (IL4), interleukin 8 (IL8), and interleukin 17 (IL17) in vaginal lavage fluid at different stages of RVVC infection. RESULTS: There were no significant differences between VVC and RVVC fungi except that the Sap activity was lower for RVVC-causing C. albicans than for VVC-causing C. albicans. C. albicans with both strong Sap and weak Sap induced RVVC in mice. C. albicans with strong Sap had a reduced RVVC infection rate. In addition, C. albicans with strong Sap stimulated the vaginal epithelium to secrete more IL4, IL8, and IL17. CONCLUSION: Compared with that of VVC-causing C. albicans, the Sap activity of RVVC-causing C. albicans was lower. C. albicans with strong Sap was less capable of causing repeated vaginal infections than that with weak Sap and stimulated the vaginal epithelium to produce more cytokines.

Direct Assessment of the Toxicity of Molybdenum Disulfide Atomically Thin Film and Microparticles via Cytotoxicity and Patch Testing.

The low toxicity of molybdenum disulfide (MoS2 ) atomically thin film and microparticles is confirmed via cytotoxicity and patch testing in this report. The toxicity of MoS2 thin film and microparticles is extensively studied but is still inconclusive due to potential organic contamination in the preparations of samples. Such contamination is avoided here through preparing MoS2 atomically thin film via direct sulfurization of molybdenum thin film on quartz plate, which permits a direct assessment of its toxicity without any contamination. Six different types of cells, including normal, cancer, and immortal cells, are cultured in the media containing MoS2 thin film on quartz plates or dispersed MoS2 microparticles and their viability is evaluated with respect to the concentrations of samples. Detached thin films from the quartz plates are also investigated to estimate the toxicity of dispersed MoS2 in biological media. Allergy testing on skin of guinea pigs is also conducted to understand their effect on animal skins. By avoiding possible organic contamination, the low toxicity of MoS2 atomically thin film and microparticles to cells and animal skins paves the way for its applications in flexible biosensing/bioimaging devices and biocompatible coatings.

Mechanism of Vaginal Epithelial Cell Pyroptosis Induced by the NLRP3 Inflammasome in Vulvovaginal Candidiasis.

PROBLEM: Vulvovaginal candidiasis (VVC) is a common mucosal fungal infection, and Candida albicans is the main causative agent. The NLRP3 inflammasome plays an important role in VVC, but the underlying mechanism is unknown. METHOD OF STUDY: Vaginal epithelial cells were divided into three groups: control, C. albicans strain SC5314 (wild-type, WT), and WT+ Matt Cooper Compound 950 (MCC950, a specific NLRP3 inhibitor). After human vaginal epithelial cells were pretreated with 1 µmol/L MCC950 for 2 h, C. albicans (MOI = 1) was cocultured with the human vaginal epithelial cells for 12 h. The cell supernatants were collected, LDH was detected, and the IL-1ß and IL-18 levels were determined by ELISA. The expression of the pyroptosis-related proteins NLRP3, Caspase-1 p20 and GSDMD was measured by Western blotting analysis. The protein expression of the pyroptosis-related N-terminus of GSDMD (GSDMD-N) was detected by immunofluorescence. RESULTS: In this study, we showed that the WT C. albicans strain induced pyroptosis in vaginal epithelial cells, as indicated by the LDH and proinflammatory cytokine levels and the upregulated levels of the pyroptosis-related proteins NLRP3, Caspase-1 p20, and GSDMD-N. MCC950 reversed the changes in the expression of these proteins and proinflammatory cytokines in vaginal epithelial cells. CONCLUSION: C. albicans activated the NLRP3 inflammasome to induce vaginal epithelial cell pyroptosis. MCC950 inhibited the NLRP3 inflammasome, reduced vaginal epithelial cell pyroptosis, and decreased the release of inflammatory cytokines.

The effect of long-term COVID-19 infection on maternal and fetal complications: a retrospective cohort study conducted at a single center in China.

Investigate the effect of long-term COVID-19 on maternal and fetal complications. A retrospective cohort study was conducted. A total of 623 pregnant women who delivered in Kunming First People's Hospital from November 1, 2022 to July 31, 2023 were selected. By employing statistical methods, we compared the associations between maternal and fetal complications in pregnant women with acute COVID-19 during pregnancy, long-term COVID-19, and non-COVID-19 pregnant women. In the final 623 samples, there were 209 pregnant women with acute COVID-19, 72 pregnant women with long-term COVID-19, and 342 pregnant women without COVID-19. The epidemiological and clinical characteristics of all subjects were similar. Pregnant individuals who developed long-term COVID-19 during their pregnancy had an increased risk of experiencing gestational hypertension (OR 3.344, 95% CI 1.544-7.243), gestational diabetes mellitus (OR 2.301, 95% CI 1.290-4.102), and fetal intrauterine growth restriction (OR 2.817, 95% CI 1.385-5.952). Multivariate binary logistic regression analysis showed that this association remained consistent even after adjusting for confounders and performing subgroup analyses. Other maternal and fetal complications, such as premature rupture of membranes, preterm delivery, neonatal asphyxia, and transfer of neonates to NICU, did not exhibit statistically significant associations. After linear regression analysis, the platelet count (ß: - 0.127, 95% CI - 0.001-0.000) of pregnant women with long-term COVID-19 was slightly lower than that of non-COVID-19 pregnant women, and the other coagulation parameters were not statistically significant. The incidence of gestational hypertension, gestational diabetes mellitus and fetal intrauterine growth restriction in pregnant women with long-term COVID-19 is significantly increased, but it does not further increase the coagulation status.

The Mechanism of IL-17 Regulating Neutrophils Participating in Host Immunity of RVVC Mice.

Vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC) are the most common lower genital tract infections in reproductive women. In recent years, the research on its pathogenesis mainly focuses on vaginal local immunity and IL-17 as key factors in adaptive immunity, attracting much attention. However, the role of IL-17 in local immunity in VVC and RVVC is poorly understood. At the same time, neutrophils are considered the most effective way to control and eliminate candidal infection and have a controversial role in VVC and RVVC. In this study, we built a mouse RVVC model. After analyzing the vaginal lavage solution of RVVC mice with an inflammatory factor antibody chip and ELISA, we found that IL-17 may play a protective role in RVVC. The experiment of constructing RVVC mice with different concentrations of IL-17 using halofuginone and comparing the vaginal fungi load and vaginal epithelial damage verified that IL-17 had a protective effect in RVVC. In addition, in vitro and in vivo studies found that IL-17 can promote neutrophil apoptosis and recruit neutrophils in the vagina. The neutrophils in the vagina can secrete IL-17 in an autocrine manner. These two may be why IL-17 plays a protective role in RVVC. In summary, the study suggests that IL-17-mediated regulation of neutrophil function is involved in host immune response to RVVC, which helps us to further understand the potential mechanism of IL-17 in RVVC.

Removal of heavy metals from fly ash using electrodialysis driven by a bioelectrochemical system: a case study of Pb, Mn, Cu and Cd.

Municipal solid waste incineration (MSWI) fly ash is classified as hazardous waste due to high leachable heavy metals, and incineration leachate belongs to organic wastewater with high biodegradability. Electrodialysis (ED) has shown potential for the removal of heavy metals from fly ash, and bioelectrochemical system (BES) employs biological and electrochemical reactions to generate electricity and remove contaminants from a wide range of substrates. In this study, the ED-BES coupled system was constructed for the co-treatment of fly ash and incineration leachate, where the ED was driven by BES. The treatment effect of fly ash by varying additional voltage, initial pH and liquid-to-solid (L/S) ratio was evaluated. Results showed that the highest removal rates of Pb, Mn, Cu and Cd were 25.43%, 20.13%, 32.14% and 18.87% after 14 days treatment of the coupled system, respectively. These values were obtained under 300 mV of additional voltage, L/S 20 and initial pH3. After the treatment of the coupled system, the fly ash leaching toxicity was lower than the threshold of GB5085.3-2007. The highest energy saving for removed Pb, Mn, Cu and Cd were 6.72, 15.61, 8.99 and 17.46 kWh/kg, respectively. The ED-BES can be considered a cleanliness approach to treating fly ash and incineration leachate simultaneously.

Analysis of whole genome-wide methylation and gene expression profiles in visceral omental adipose tissue of pregnancies with gestational diabetes mellitus.

BACKGROUND: DNA methylation is the most extensively studied epigenetic modification which had been suspected to be involved in the progress of gestational diabetes mellitus（GDM）. It is vital to investigate the expression profile and methylation profile in the GDM adipose tissue samples to learn more about the relationship between the two profiles. METHODS: Illumina Human Methylation 450 k DNA Analysis Beadchip and whole Human Gene Expression Array were selected to screen for methylation and gene expression in the omental visceral adipose tissue of pregnant women. Validation of methylation of DMGs was conducted by bisulfate pyrosequencing and expression of DEGs by q RT-PCR. RESULTS: Global gene methylation profiling and whole genome expression profiling were conducted in visceral omental adipose tissue (VOAT) between GDM and normal pregnancies. Compared with controls, 935 genes were commonly dysregulated in the GDM group, including 450 down-regulated DEGs and 485 up-regulated DEGs. The Seven overlapping genes between DEGs and DMGs were extracted, including C10orf10, FSTL1, GSTT1, HLA-DPB1, HLA-DRB5, HSPA6 and MSLN. Among them, C10orf10, FSTL1, GSTT1, HLA-DPB1, HLA-DRB5 showed hypermethylation and up-regulated expression, while HSPA6 show hypomethylation and down-regulated expression. Typical negative correlation between gene expression and DNA methylation level was only found in MSLN with significant hypermethylation in the CpG island and downregulated transcription. No gene was found to be significantly hypomethylated in the CpG islands and unregulated transcription. CONCLUSION: We found that antigen processing and presentation pathway and immune-related genes were closely associated with gestational diabetes mellitus in the visceral omental adipose tissue of Chinese pregnant women, based on the integration analysis of expression and methylation profiles. These results may be valuable for the prognostic biomarkers and future therapeutic targets.

Antibody-free detection of protein phosphorylation using intrinsic peroxidase-like activity of platinum/carbon dot hybrid nanoparticles.

Platinum and carbon dot hybrid nanomaterials are prepared for visualized detection of phosphoproteins without the need for antibodies or enzymes. This new strategy can be used for colorimetric detection of phosphoproteins induced by protein kinase as well as protein phosphorylation sites on cell membranes.

A versatile quantitation platform based on platinum nanoparticles incorporated volumetric bar-chart chip for highly sensitive assays.

Platinum nanoparticles incorporated volumetric bar-chart chip (PtNPs-V-Chip) is able to be used for point-of-care tests by providing quantitative and visualized readout without any assistance from instruments, data processing, or graphic plotting. To improve the sensitivity of PtNPs-V-Chip, hybridization chain reaction was employed in this quantitation platform for highly sensitive assays that can detect as low as 16 pM Ebola Virus DNA, 0.01ng/mL carcinoembryonic antigen (CEA), and the 10 HER2-expressing cancer cells. Based on this amplified strategy, a 100-fold decrease of detection limit was achieved for DNA by improving the number of platinum nanoparticle catalyst for the captured analyte. This quantitation platform can also distinguish single base mismatch of DNA hybridization and observe the concentration threshold of CEA. The new strategy lays the foundation for this quantitation platform to be applied in forensic analysis, biothreat detection, clinical diagnostics and drug screening.

Nanoporous Glass Integrated in Volumetric Bar-Chart Chip for Point-of-Care Diagnostics of Non-Small Cell Lung Cancer.

Point-of-care (POC) testing has the potential to enable rapid, low-cost, and large-scale screening. POC detection of a multiplexed biomarker panel can facilitate the early diagnosis of non-small cell lung cancer (NSCLC) and, thus, may allow for more timely surgical intervention for life-saving treatment. Herein, we report the nanoporous glass (NPG) integrated volumetric bar-chart chip (V-Chip) for POC detection of the three NSCLC biomarkers CEA, CYFRA 21-1, and SCCA, by the naked eye. The 3D nanostructures in the NPG membrane efficiently increase the number of binding sites for antibodies and decrease the diffusion distance between antibody and antigen, enabling the low detection limit and rapid analysis time of the NPG-V-Chip. We utilized the NPG-V-Chip to test the NSCLC biomarker panel and found that the limit of detection can reach 50 pg/mL (10-fold improvement over the original V-Chip), and the total assay time can be decreased from 4 to 0.5 h. We then detected CEA in 21 serum samples from patients with common cancers, and the on-chip results showed good correlation with the clinical results. We further assayed 10 lung cancer samples using the device and confirmed the results obtained using conventional ELISA methods. In summary, the NPG-V-Chip platform has the ability of multiplex, low detection limit, low cost, lack of need for accessory equipment, and rapid analysis time, which may render the V-Chip a useful platform for quantitative POC detection in resource-limited settings and personalized diagnostics.

Integrative volumetric bar-chart chip for rapid and quantitative point-of-care detection of myocardial infarction biomarkers.

Here we developed an integrated volumetric bar-chart chip (IV-Chip) technology by integration of our previous V-Chip with a fluid handling design to generate an instrument-free POC device and greatly reduce the detection time and effort. The IV-Chip test requires only 1 µL of serum separated from finger-prick blood. The serum sample and ELISA reagents are directly loaded into the device using a pipette, and a shift of the two layers of the device generates homogeneous liquid segments in the microfluidic channel. Under vacuum pressure generated by a regular syringe, the segments flow into the ELISA wells in sequence and a sandwich ELISA reaction takes place. As a result of the automated washing and reacting strategy, the IV-Chip allows rapid tests for myocardial infarction biomarkers, and turnaround time is greatly reduced to 15 min. The specificity and accuracy of quantitative multiplex detection of MI biomarkers CK-MB, troponin I and myoglobin, are 87.5% and 95.8%, respectively.

[Study on correlation between histocompatibility leukocyte antigen class II gene and gestational diabetes mellitus].

OBJECTIVE: To investigate correlation between part of histocompatibility leukocyte antigen (HLA) class II gene and gestational diabetes mellitus (GDM). METHODS: Part of HLA-II gene was examined by polymerase chain reaction sequence specific primer (PCR-SSP). Forty-eight GDM women were served as study group, and forty-eight normal pregnant women were selected as control group. RESULTS: The frequencies of DRB1*0301, DRB1*1302 and DQA1*0301 of GDM (18.8%, 45.8% and 64.6%) in study group were significantly increased compared with control group (4.2%, 18.8% and 41.7%) (P < 0.05). CONCLUSIONS: The results suggest that there is correlation between HLA class II gene and gestational diabetes mellitus. DRB1*0301, DRB1*1302 and DQA1*0301 are the susceptible alleles of GDM.

Luminescent lanthanide graphene for detection of bacterial spores and cysteine.

Here, we describe a new approach for preparation of luminescent lanthanide graphene in the presence of dipicolinic acid (DPA). Hg(2+) can competitively bind with DPA which greatly quenches the fluorescence and the resultant complex is able to selectively and sensitively detect cysteine with a detection limit of 5 nM.