HGF facilitates methylation of MEG3, potentially implicated in vemurafenib resistance in melanoma.

BACKGROUND: Melanoma, a frequently encountered cutaneous malignancy characterized by a poor prognosis, persists in presenting formidable challenges despite the advancement in molecularly targeted drugs designed to improve survival rates significantly. Unfortunately, as more therapeutic choices have developed over time, the gradual emergence of drug resistance has become a notable impediment to the effectiveness of these therapeutic interventions. The hepatocyte growth factor (HGF)/c-met signaling pathway has attracted considerable attention, associated with drug resistance stemming from multiple potential mutations within the c-met gene. The activation of the HGF/c-met pathway operates in an autocrine manner in melanoma. Notably, a key player in the regulatory orchestration of HGF/c-met activation is the long non-coding RNA MEG3. METHODS: Melanoma tissues were collected to measure MEG3 expression. In vitro validation was performed on MEG3 to prove its oncogenic roles. Bioinformatic analyses were conducted on the TCGA database to build the MEG3-related score. The immune characteristics and mutation features of the MEG3-related score were explored. RESULTS: We revealed a negative correlation between HGF and MEG3. In melanoma cells, HGF inhibited MEG3 expression by augmenting the methylation of the MEG3 promoter. Significantly, MEG3 exhibits a suppressive impact on the proliferation and migration of melanoma cells, concurrently inhibiting c-met expression. Moreover, a predictive model centered around MEG3 demonstrates notable efficacy in forecasting critical prognostic indicators, immunological profiles, and mutation statuses among melanoma patients. CONCLUSIONS: The present study highlights the potential of MEG3 as a pivotal regulator of c-met, establishing it as a promising candidate for targeted drug development in the ongoing pursuit of effective therapeutic interventions.

Variations in Breast Milk Iodine Concentration over 24 h among Lactating Women in Northern China.

BACKGROUND: Adequate breast milk iodine concentration (BMIC) is essential for the growth and cognitive development of exclusively breastfed infants; however, data on variations in BMIC over 24 h are limited. OBJECTIVE: We aimed to explore in lactating women the variation in 24-h BMIC. METHODS: Thirty pairs of mothers and breastfed infants aged 0-6 mo were recruited from the cities of Tianjin and Luoyang, China. A 3-d 24-h dietary record, including salt intake, was performed to assess the dietary iodine intake of lactating women. Breast milk samples before and after each feeding for 24 h and 24-h urine samples were collected from the women for 3 d to estimate iodine excretion. A multivariate linear regression model was used to analyze the factors influencing BMIC. A total of 2658 breast milk samples and 90 24-h urine samples were collected. RESULTS: The median BMIC and 24-h urine iodine concentration (UIC) of lactating women for a mean of 3.6 ± 1.48 mo were 158 µg/L and 137 µg/L, respectively. The interindividual variability of BMIC (35.1%) was higher than that observed within individuals (11.8%). The variation in BMIC showed a "V" shaped curve over 24 h. The median BMIC at 08:00-12:00 (137 µg/L) was significantly lower than that at 20:00-24:00 (163 µg/L) and 00:00-04:00 (164 µg/L). A progressively increasing curve was obtained for BMIC until it peaked at 20:00 and plateaued at a higher concentration from 20:00 to 04:00 than at 08:00-12:00 (all P < 0.05). BMIC was associated with dietary iodine intake (ß: 0.366; 95% CI: 0.004, 0.018) and infant age (ß: -0.432; 95% CI: -1.07, -0.322). CONCLUSIONS: Our study shows that the BMIC presents a "V" shaped curve over 24 h. We recommend that breast milk samples be collected between 08:00 and 12:00 for evaluation of the iodine status of lactating women.

Construction of stable membranal CMTM6-PD-L1 full-length complex to evaluate the PD-1/PD-L1-CMTM6 interaction and develop anti-tumor anti-CMTM6 nanobody.

CKLF (chemokine-like factor)-MARVEL transmembrane domain containing protein 6 (CMTM6) is a novel regulator to maintain the stability of PD-L1. CMTM6 can colocalize and interact with PD-L1 on the recycling endosomes and cell membrane, preventing PD-L1 from lysosome-mediated degradation and proteasome-mediated degradation thus increasing the half-life of PD-L1 on the cell membrane. The difficulties in obtaining stable full-length PD-L1 and CMTM6 proteins hinder the research on their structures, function as well as related drug development. Using lauryl maltose neopentyl glycol (LMNG) as the optimized detergent and a cell membrane mimetic strategy, we assembled a stable membrane-bound full-length CMTM6-PD-L1 complex with amphipol A8-35. When the PD-1/PD-L1-CMTM6 interactions were analyzed, we found that CMTM6 greatly enhanced the binding and delayed the dissociation of PD-1/PD-L1, thus affecting immunosuppressive signaling and anti-apoptotic signaling. We then used the CMTM6-PD-L1 complex as immunogens to generate immune repertoires in camels, and identified a functional anti-CMTM6 nanobody, called 1A5. We demonstrated that the anti-CMTM6 nanobody greatly decreased T-cell immunosuppression and promoted apoptotic susceptibility of tumor cells in vitro, and mainly relied on the cytotoxic effect of CD8+ T-cells to exert tumor growth inhibitory effects in CT26 tumor-bearing mice. In conclusion, the stable membrane-bound full-length CMTM6-PD-L1 complex has been successfully used in studying PD-1/PD-L1-CMTM6 interactions and CMTM6-targeting drug development, suggesting CMTM6 as a novel tumor immunotherapy target.

The associations of gestational weight gain and midpregnancy lipid levels with placental size and placental-to-birth weight ratio: findings from a chinese birth cohort study.

BACKGROUND: The placenta serves as the sole maternal organ responsible for transmitting nutrients to the fetus, playing a crucial role in supporting standard fetal growth and development. To date, only a small number of studies have investigated the impact of maternal gestational weight gain and lipid concentrations on placental development. This study aimed to explore the influence of weight gain during pregnancy and lipid levels in the second trimester on placental weight, volume, and the placental weight ratio. METHODS: This birth cohort study encompassed 1,358 mother-child pairs. Placental data for each participant was gathered immediately post-delivery, and the study incorporated data on gestational weight gain throughout pregnancy and lipid profiles from the mid-trimester. A linear regression model was employed to assess the correlations between gestational weight gain, mid-trimester lipid levels, and metrics such as placental weight, placental volume, and the placental-to-birth weight ratio (PFR). RESULTS: In the study groups of pre-pregnancy underweight, normal weight, and overweight, the placental weight increased by 4.93 g (95% CI: 1.04-8.81), 2.52 g (95% CI: 1.04-3.99), and 3.30 g (95% CI: 0.38-6.22) per 1 kg of gestational weight gain, respectively. Within the pre-pregnancy underweight and normal weight groups, the placental volume increased by 6.79 cm^3 (95% CI: 3.43-10.15) and 2.85 cm^3 (95% CI: 1.31-4.39) per 1 kg of gestational weight gain, respectively. Additionally, placental weight exhibited a positive correlation with triglyceride (TG) levels (ß = 9.81, 95% CI: 3.28-16.34) and a negative correlation with high-density lipoprotein (HDL-C) levels (ß = - 46.30, 95% CI: - 69.49 to - 23.11). Placental volume also showed a positive association with TG levels (ß = 14.54, 95% CI: 7.69-21.39). Conversely, PFR demonstrated a negative correlation with increasing HDL-C levels (ß = - 0.89, 95% CI: - 1.50 to - 0.27). CONCLUSIONS: Gestational weight gain was significantly correlated with both placental weight and volume. This association was especially pronounced in women who, prior to pregnancy, were underweight or of normal weight. Additionally, TG and HDL-C levels during the mid-trimester were linked to placental development.

BAP1 antagonizes WWP1-mediated transcription factor KLF5 ubiquitination and inhibits autophagy to promote melanoma progression.

Accumulating evidence revealed the abnormal expression of KLF5 in human cancers while its role in melanoma remains uncharacterized. This study aimed to explore the role of KLF5 in the proliferation and metastasis of melanoma. Bioinformatics analysis was performed to detect WWP1, BAP1 and KLF5 expression in melanoma, followed by expression determination on clinical tissues from melanoma patients and cancer cells. The cancer cells were infected with lentivirus expressing KLF5 or BAP1 while PI3K, AKT and mTOR expression was detected and autophagy was observed. Treated cells were injected to mice when tumor growth was measured and autophagy-related protein was detected. Plasmids expressing WWP1 and Ub-K48 were co-transfected into treated melanoma cells while immunoprecipitation assay was performed to determine the interaction among KLF5, WWP1, and BAP1. WWP1 was poorly expressed in melanoma cells and tissues whereas KLF5 was highly expressed and was positively correlated to poor prognosis. KLF5 promoted melanoma cell malignant phenotypes as well as inhibited autophagy. Interestingly, KLF5 contributed to activation of PI3K-AKT-mTOR signaling pathway, thereby inhibiting autophagy in melanoma cells. WWP1 mediated K48-linked ubiquitination of KLF5 to promote its degradation, and BAP1 antagonized this modification and stabilized KLF5 protein expression. Besides, BAP1 promoted KLF5-mediated growth of melanoma in vivo. Taken altogether, BAP1 antagonized WWP1-mediated ubiquitination of KLF5 to inhibit autophagy and promote melanoma development.

NIP1;2 is a plasma membrane-localized transporter mediating aluminum uptake, translocation, and tolerance in Arabidopsis.

Members of the aquaporin (AQP) family have been suggested to transport aluminum (Al) in plants; however, the Al form transported by AQPs and the roles of AQPs in Al tolerance remain elusive. Here we report that NIP1;2, a plasma membrane-localized member of the Arabidopsis nodulin 26-like intrinsic protein (NIP) subfamily of the AQP family, facilitates Al-malate transport from the root cell wall into the root symplasm, with subsequent Al xylem loading and root-to-shoot translocation, which are critical steps in an internal Al tolerance mechanism in Arabidopsis We found that NIP1;2 transcripts are expressed mainly in the root tips, and that this expression is enhanced by Al but not by other metal stresses. Mutations in NIP1;2 lead to hyperaccumulation of toxic Al3+ in the root cell wall, inhibition of root-to-shoot Al translocation, and a significant reduction in Al tolerance. NIP1;2 facilitates the transport of Al-malate, but not Al3+ ions, in both yeast and Arabidopsis We demonstrate that the formation of the Al-malate complex in the root tip apoplast is a prerequisite for NIP1;2-mediated Al removal from the root cell wall, and that this requires a functional root malate exudation system mediated by the Al-activated malate transporter, ALMT1. Taken together, these findings reveal a critical linkage between the previously identified Al exclusion mechanism based on root malate release and an internal Al tolerance mechanism identified here through the coordinated function of NIP1;2 and ALMT1, which is required for Al removal from the root cell wall, root-to-shoot Al translocation, and overall Al tolerance in Arabidopsis.

Community and individual iodine status assessment in premenopausal women in Shanxi, China: Repeated spot urine versus 24-hour urine.

BACKGROUND AND OBJECTIVES: Urinary iodine is an essential index of iodine nutrition evaluation. To establish the number of repeated spot urine collections necessary to reflect individual iodine status over 2 months and assess its feasibility to serve as an independent indicator of individual iodine status. METHODS AND STUDY DESIGN: We performed a longitudinal, 2-months study from May to June in 2015 of 23 apparently healthy Chinese women aged 18 to 44 (32±9) y. Spot urine samples were collected on any two days of the week, and 24-h urine samples were collected once every 6 days. RESULTS: 368 spot urine and 230 24-h urine samples were analysed. The median urinary iodine concentration (UIC) was 140.5 (75.2, 246.9) µg/L. The estimated 24-h urinary iodine excretion (24-h UIE) values from spot urine samples and measured 24-h UIE values from 24-h urine samples were 348±240 µg/24h and 330±216 µg/24h (p=0.003), respectively. Irrespective of the urinary iodine method, the intra- individual coefficient of variation (CV%) was lower than the inter-individual CV%. Bland-Altman analysis revealed differences between spot urine and 24-h urine. When the precision ranges with 95% confidence were ±15%, ±20%, ±25% or 30%, the number of an individual's spot urine samples required were 30, 16, 11 or 8, respectively. CONCLUSIONS: Repeated spot urine is not a feasible way to assess recent individual iodine intake. The development of a multi-indicator system could provide an acceptable individual evaluation index of iodine status.

Silencing of microRNA-27a facilitates autophagy and apoptosis of melanoma cells through the activation of the SYK-dependent mTOR signaling pathway.

Melanoma is considered as an aggressive neoplastic transformation and featured with high metastatic potential. Although some studies have provided targets for novel therapeutic interventions, clinical development of targeted drugs for melanoma still remains obscure. Therefore, this study aims to identify the role of microRNA-27a (miR-27a) in autophagy and apoptosis of melanoma cells in regulating spleen tyrosine kinase (SYK)-mediated the mammalian target of rapamycin (mTOR) signaling pathway. A microarray-based analysis was made to screen differentially expressed genes and predict target miRNA. Melanoma specimens were collected with pigmented nevus as a control. Melanoma cell line Mel-RM was treated with miR-27a inhibitor or pcDNA-SYK to prove their effects on autophagy and apoptosis of melanoma cells. The volume change and tumor mass of nude mice in each group were detected by the tumorigenesis assay. Microarray-based analysis results showed that SYK was lowly expressed in melanoma cells and may be regulated by miR-27a. Besides, miR-27a expression was increased whereas SYK expression was decreased in melanoma tissues. Meanwhile, miR-27a was positively correlated with tumor stage and lymph node metastasis of melanoma tissues. Furthermore, miR-27a targeted SYK and silencing of miR-27a or overexpression of SYK cells promoted autophagy and apoptosis of melanoma cells and reduced their tumorigenic ability in vivo. In conclusion, this study proves that silencing of miR-27a facilitates autophagy and apoptosis of melanoma cells by upregulating SYK expression and activating the mTOR signaling pathway. The finding offers new ideas for the clinical development of melanoma.

Natural variation underlies alterations in Nramp aluminum transporter (NRAT1) expression and function that play a key role in rice aluminum tolerance.

Aluminum (Al) toxicity is a major constraint for crop production on acid soils which compose ∼ 40% of arable land in the tropics and subtropics. Rice is the most Al-tolerant cereal crop and offers a good model for identifying Al tolerance genes and mechanisms. Here we investigated natural variation in the rice Nramp aluminum transporter (NRAT1) gene encoding a root plasma membrane Al uptake transporter previously hypothesized to underlie a unique Al tolerance mechanism. DNA sequence variation in the NRAT1 coding and regulatory regions was associated with changes in NRAT1 expression and NRAT1 Al transport properties. These sequence changes resulted in significant differences in Al tolerance that were found to be associated with changes in the Al content of root cell wall and cell sap in 24 representative rice lines from a rice association panel. Expression of the tolerant OsNRAT1 allele in yeast resulted in higher Al uptake than did the sensitive allele and conferred greater Al tolerance when expressed in transgenic Arabidopsis. These findings indicate that NRAT1 plays an important role in rice Al tolerance by reducing the level of toxic Al in the root cell wall and transporting Al into the root cell, where it is ultimately sequestered in the vacuole. Given its ability to enhance Al tolerance in rice and Arabidopsis, this work suggests that the NRAT1 gene or its orthologs may be useful tools for enhancing Al tolerance in a wide range of plant species.

Prevalence and associated factors of induced abortion among rural married women: a cross-sectional survey in Anhui, China.

AIM: This study aims to assess the prevalence of and factors associated with induced abortion among married women in rural areas of Anhui Province, China. MATERIAL AND METHODS: A multistage probability sampling method was used to identify a representative sample of 53,652 married women aged 18-49 years in rural areas of Anhui Province, China. All women were interviewed in the form of a standardized questionnaire. RESULTS: We found that 32.0% (16,800) of these women had had at least one induced abortion: 21.1% (11,090) of women had had one; 7.6% (3976) of women had had two; and 4.1% (1734) of women had had at least three. The number of induced abortions per 100 pregnancies was found to be 22.0. Multivariate analysis showed that education, the age of a woman at her first marriage, number of total births, number of total pregnancies, and contraceptive methods were significant predictors for induced abortion after controlling for women's current age, employment and family yearly income. CONCLUSION: The study shows that the prevalence of induced abortion is still very high among married women in rural China, and highly effective methods of contraception (sterilization, intrauterine device) decrease women's recourse to induced abortion.

A systematic review and quantitative assessment of sleep-disordered breathing during pregnancy and perinatal outcomes.

PURPOSE: Previous investigations have suggested a strong association between sleep-disordered breathing (SDB) during pregnancy and perinatal outcomes. However, the results of the following replication studies were not always concordant. Therefore, this meta-analysis was conducted to evaluate the more reliable estimate. METHODS: A systematic literature search was performed on PubMed, Springer Link, and EMBASE to identify all eligible studies published before August 2013. Summary odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using fixed or random effects model. RESULTS: A total of 24 publications met the inclusion criteria and were included in this meta-analysis. Findings demonstrated that moderate-to-severe SDB during pregnancy was associated with gestational diabetes mellitus (OR=1.78; 95% CI, 1.29 to 2.46), pregnancy-related hypertension (OR=2.38; 95% CI, 1.63 to 3.47), preeclampsia (OR=2.19; 95% CI, 1.71 to 2.80), preterm delivery (OR=1.98; 95% CI, 1.59 to 2.48), low birth weight (OR=1.75; 95% CI, 1.33 to 2.32), neonatal intensive care unit (NICU) admission (OR=2.43; 95% CI, 1.61 to 3.68), intrauterine growth restriction (OR=1.44; 95% CI, 1.22 to 1.71), and Apgar score of <7 at 1 min (OR=1.78; 95% CI, 1.10 to 2.91) based on all studies but not gestational age and birth weight. CONCLUSIONS: This meta-analysis revealed that moderate-to-severe SDB during pregnancy may be associated with most of adverse perinatal outcomes. Further well-designed studies are warranted to confirm our findings.

Deciphering the tumor-suppressive role of PSMB9 in melanoma through multi-omics and single-cell transcriptome analyses.

Skin cutaneous melanoma (SKCM) poses a significant challenge in skin cancers. Recent immunotherapy breakthroughs have revolutionized melanoma treamtment, yet tumor heterogeneity persists as an obstacle. Epigenetic modifications orchestrated by DNA methylation contributed to tumorigenesis, thus potentially unveiling melanoma prognosis. Here, we identified an interferon-gamma (IFN-g) sensitive subtype, which possesses favorable outcomes, robust infiltration CD8+T cells, and IFN-g score in bulk RNA-seq profile. Subsequently, we established an IFN-g sensitivity signature based on machine learning. We validated that PSMB9 is strongly correlated with immunotherapy response in both methylation and expression cohorts in this 10-probe signature. We assumed that PSMB9 acts as a putative melanoma suppressor, for its activation of CD8+T cell; capacity to modulate IFN-γ secretion; and dynamics altering IFN-g receptors in bulk tissue. We performed single-cell RNA-seq on immunotherapy patients' tissue to uncover the nuanced role of PSMB9 in activating CD8T + cells, enhancing IFN-g, and influencing malignant cells receptors and transcriptional factors. Overexpress PSMB9 in two SKCM cell lines to mimic the hypomethylated state to approve our conjecture. Strong cell proliferation and migration inhibition were detected on both cells, indicating that PSMB9 is present in tumor cells and that high expression is detrimental to tumor growth and migration. Overall, comprehensive integrated analysis shows that PSMB9 emerges as a vital prognostic marker, acting predictive potential regarding immunotherapy in melanoma. This evidence not only reveals the multifaceted impact of PSMB9 on both malignant and immune cells but also serves as a prospective target for undergoing immunotherapeutic strategies in the future.

MicroRNA-124 plays an inhibitory role in cutaneous squamous cell carcinoma cells via targeting SNAI2, an immunotherapy determinant.

MicroRNAs (miRs) play multiple roles during cutaneous squamous cell carcinoma (CSCC) progression. Previous studies suggest miR-124 could inhibit cancer development in CSCC. METHODS: Obtained 63 pairs of CSCC and adjacent tissues for analysis. Cultured HaCaT and two CSCC cell lines (A431 and SCL-1) in DMEM (10 % FBS). Transfected cells using Lipofectamine 2000 with various miR-124 mimics, inhibitors, or Snail family transcriptional repressor 2 (SNAI2) expression plasmid. Performed a series of assays, including real-time quantitative PCR, Western blot, CCK8, wound healing, transwell, and luciferase reporter gene assay, to examine the effects of miR-124 on CSCC cells. RESULTS: An evident downregulation of miR-124 in CSCC tissues, which was related to advanced disease stage and nodal metastasis. Overexpressing miR-124 could reduce the proliferation, migration, and invasion abilities of CSCC cells. It was verified that miR-124 targets the SNAI2 in CSCC cells. Moreover, ectopic expression of SNAI2 rescued the suppressive effects on CSCC cells induced by miR-124 overexpression. Furthermore, miR-124 increased cell sensitivity to cisplatin. Besides, SNAI2 is a critical factor in the immune-related aspects of CSCC and its modulation may influence the response to immunotherapy. CONCLUSION: We demonstrate that miR-124 inhibits CSCC progression through downregulating SNAI2, and thus it may be a molecular candidate for treating CSCC in the clinic.

Ultrasound-assisted deep eutectic solvents extraction of polysaccharides from Loquat leaf: Process optimization and bioactivity study.

Loquat leaves are the by-product of loquat fruit production. Polysaccharides are one of the main active ingredients in loquat leaves. In this study, polysaccharides were extracted from loquat leaves by ultrasonic-assisted deep eutectic solvents (DESs) extraction method. Further, the extracted crude loquat leaf polysaccharides (CLLP) were purified and separated via S-8 resin and DEAE-52 cellulose column chromatography, respectively. Additionally, the effects of polysaccharides on activity of sperm in boar semen preserved in medium at 17 °C, were evaluated preliminarily. DES, composed of choline chloride/ethylene glycol (1:6, molar ratio), was proved to be the suitable solvent for LLP extraction. The optimized extraction conditions were water content 44 %, liquid-solid ratio 1:29 (g/g), extraction temperature 61 °C and extraction time 98 min. Under these conditions, the LLP yield was 57.82 ± 1.50 mg/g. A homogeneous polysaccharide (LLP1-2, Mw: 2.17 × 104 Da) was isolated from CLLP. Its total sugar, uronic acid and protein contents were 76.31 ± 1.25 %, 14.19 ± 0.67 % and 3.28 ± 0.42 %, respectively. Further, 800 µg/mL LLP1-2 could effectively enhance the antioxidant activity of sperm. This study laid a foundation for DESs and column chromatography in the field of polysaccharide extraction and separation, proving that LLP can be used as a natural antioxidant for sperm preservation.

A promoter-swap strategy between the AtALMT and AtMATE genes increased Arabidopsis aluminum resistance and improved carbon-use efficiency for aluminum resistance.

The primary mechanism of Arabidopsis aluminum (Al) resistance is based on root Al exclusion, resulting from Al-activated root exudation of the Al(3+) -chelating organic acids, malate and citrate. Root malate exudation is the major contributor to Arabidopsis Al resistance, and is conferred by expression of AtALMT1, which encodes the root malate transporter. Root citrate exudation plays a smaller but still significant role in Arabidopsis Al resistance, and is conferred by expression of AtMATE, which encodes the root citrate transporter. In this study, we demonstrate that levels of Al-activated root organic acid exudation are closely correlated with expression of the organic acid transporter genes AtALMT1 and AtMATE. We also found that the AtALMT1 promoter confers a significantly higher level of gene expression than the AtMATE promoter. Analysis of AtALMT1 and AtMATE tissue- and cell-specific expression based on stable expression of promoter-reporter gene constructs showed that the two genes are expressed in complementary root regions: AtALMT1 is expressed in the root apices, while AtMATE is expressed in the mature portions of the roots. As citrate is a much more effective chelator of Al(3+) than malate, we used a promoter-swap strategy to test whether root tip-localized expression of the AtMATE coding region driven by the stronger AtALMT1 promoter (AtALMT1(P)::AtMATE) resulted in increased Arabidopsis Al resistance. Our results indicate that expression of AtALMT1(P)::AtMATE not only significantly increased Al resistance of the transgenic plants, but also enhanced carbon-use efficiency for Al resistance.

Effect of Aging on Healing Capacity of Bituminous Composites Containing Polyphosphoric Acid.

This study examines how aging affects the healing capacity of bituminous composites containing polyphosphoric acid (PPA). PPA is commonly used in bituminous composites to enhance its elasticity, however, PPA effectiveness highly depends on other constituents on the matrix and the environmental (internal and external) factors. In terms of internal factors, the interplay between PPA and various bitumen modifiers have been extensively studied. Here, we study how external factors such as exposure to ultraviolet radiation affect PPA's efficacy, measured in terms of change in bitumen's healing index. The study results showed that the introduction of PPA to bituminous composites significantly increases the bitumen healing index, however, the change in the healing index becomes less pronounced as aging progresses. The presence of additives such as taconite were found to affect the effect of PPA on bitumen's healing index. For instance, bitumens containing 30% taconite showed the highest increase in their healing index in the presence of PPA among studied scenarios. Overall, bitumen containing PPA had a higher healing index than those without PPA regardless of the extent of aging and dosage of modifiers. This, in turn, indicates that PPA is highly effective for enhancing bitumen healing. This can be attributed to the role of PPA in promoting intermolecular interactions within the bitumen matrix.

[Annexin A5 gene polymorphism (-1C/T) and the susceptibility to pneumoconiosis in coal works].

OBJECTIVE: To explore the role of -1C/T single nucleotide polymorphism within Annexin A5 gene in the genetic susceptibility to coal worker's pneumoconiosis (CWP). METHODS: Four hundred and seventy CWP Han chinese patients and 428 Han chinese controls were enclosed in present case-control study. All subjects were exposed to coal dusts. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the -1C/T SNP in Annexin A5 gene for all subjects. The relationship between the -1C/T SNP in Annexin A5 gene and CWP was analyzed. RESULTS: CT/TT genotype in -1C/T SNP was associated with a significantly decreased risk of CWP, as compared with the CC genotype among subgroups exposed to coal dusts for ≥ 27 years (adjusted OR = 0.65, 95%CI: 0.44 - 0.98, P = 0.039) and patients with CWP at stage II (adjusted OR = 0.55, 95%CI: 0.34 - 0.90, P = 0.028). CONCLUSION: The results of present study suggest that the Annexin A5 -1C/T polymorphism may be involved in the development of CWP in Han Chinese coal miners.

The Expression and Clinical Significance of Sphingosine Kinase 1 and Vascular Endothelial Growth Factor in Endometrial Carcinoma.

The aim of the study is to investigate the expression of sphingosine kinase 1 (SPHK1) and vascular endothelial growth factor (VEGF) in patients with endometrial carcinoma and its clinical significance. The tissues of 86 cases of patients with endometrial carcinoma and 54 cases of patients with endometrial atypical hyperplasia were collected. The expression of SPHK1 and VEGF in the tissue was detected by immunohistochemistry. The expression of SPHK1 in patients with endometrial carcinoma was compared with the clinicopathological data. Results. 69 cases (82.1%) of endometrial carcinoma were positive for SPHK1, which was higher than 2 cases (3.7%) of endometrial atypical hyperplasia (P < 0.05). The VEGF expression in 54 patients (62.8%) with endometrial carcinoma was higher than that in 12 patients with endometrial atypical hyperplasia (22.2%) (P < 0.05). There was a positive correlation between SPHK1 and VEGF expressions in endometrial carcinoma (c = 0.595). The expression of SPHK1 in endometrial cancer patients was different in different pathological types, FIGO stages, lymph node metastasis, ER, and PR positive or not, and the difference between the two groups was significant (P < 0.05). There was no difference in age, degree of differentiation, and depth of myometrial infiltration (P < 0.05). The expression of SPHK1 in patients with endometrial carcinoma is increased, which is helpful for early detection of patients with endometrial carcinoma, and may play a synergistic role with VEGF in the pathogenesis and development of endometrial carcinoma.

Serum Soluble ST2 Is a Valuable Prognostic Biomarker in Patients With Acute Heart Failure.

BACKGROUND: This study aimed to investigate the clinical utility of different soluble suppression of tumorigenicity 2 (sST2) levels in assessing the severity and prognosis of patients with acute heart failure (AHF). METHODS: This was a prospective cohort study. Three hundred and thirty-one consecutively enrolled AHF patients from March 2018 to November 2019 were divided into 3 subgroups according to sST2 levels: T1 (1.15-7.70 ng/ml; N = 110), T2 (7.71-17.24 ng/ml; N = 111), and T3 (17.26-47.42 ng/ml; N = 110). The patients were followed up for a median period of 21.0 months for the development of the primary endpoint. Cox proportional hazards model was performed to evaluate the prognostic value of sST2 for the clinical outcomes. RESULTS: The mean age of patients was 69 years (range, 34-93 years), and 70.4% were male. During the follow-up period, 63 participants died. Patients with higher sST2 levels had lower left ventricular ejection fraction (correlation = -0.119, P = 0.031), and higher New York Heart Association classification (correlation = 0.443, P < 0.001) and N-terminal pro-B type natriuretic peptide (NT-proBNP) levels (correlation = 0.392, P < 0.001). Higher sST2 was also associated with creatinine, urea nitrogen, hemoglobin, and left ventricular mass index. Multivariate analysis revealed that sST2 (per log unit, hazard ratio: 2.174, 95% confidence interval [CI] 1.012-4.67, P = 0.047) and NT-proBNP (per log unit, HR 2.171, 95%CI 1.169-4.032, P < 0.001) were independent risk factors for the primary outcome in all patients with AHF. CONCLUSION: sST2 can provide prognostic information in AHF. The higher the sST2 level in patients with AHF, the higher the incidence of cardiovascular death.

PD-L1/TLR7 dual-targeting nanobody-drug conjugate mediates potent tumor regression via elevating tumor immunogenicity in a host-expressed PD-L1 bias-dependent way.

BACKGROUND: Various tumors are insensitive to immune checkpoint blockade (ICB) therapy. Toll-like receptors (TLRs) establish the link between innate and adaptive immunity, which can assist T-cell activation and serve as promising targets for combination to enhance ICB therapy. Here, we aimed to improve efficacy for anti-programmed death ligand 1 (PD-L1) therapy by developing a PD-L1/TLR7 dual-targeting nanobody-drug conjugate (NDC), based on the PD-L1 nanobodies and TLR7 agonist we developed. METHODS: PD-L1 nanobodies were obtained by phage display screening and identified through T-cell activation bioassay, in vivo imaging and quantitative biodistribution study. Immune activation and PD-L1-inducing of TLR7 agonists were evaluated in diverse innate cell models. We constructed PD-L1/TLR7 dual-targeting NDCs by chemically coupling PD-L1 nanobodies and TLR7 agonists. The antitumor effect was evaluated via several murine or humanized solid tumor models. Immunophenotyping, immune cell depletion, tumor rechallenge, RNA sequencing and PD-L1-deficient models were combined to determine the mechanism for NDCs function. The dynamics of the in vivo behaviors of NDCs were assessed based on multiorgan changes in PD-L1 levels. RESULTS: The screened PD-L1 nanobodies were characterized as tumor-targeting and alleviated T-cell immunosuppression. The TLR7 agonists induced broad innate immune responses and intratumoral PD-L1 expression on antigen-presenting cells (APCs), and its antitumor effect was dependent on intratumoral delivery. The combination of TLR7 agonists and PD-L1 nanobodies activated both innate and adaptive immunity and upregulated PD-L1-related signaling pathways. After coupling to form dual-targeting NDCs, TLR7 agonists and PD-L1 nanobodies exerted synergistic antitumor effects and safety in either 'hot' or 'cold' tumor and early or advanced tumor models, reshaped the tumor immune microenvironment and induced antitumor immune memory. CD8+ T cells and natural killer cells were the main effector cells for NDCs to function. NDCs can promote PD-L1 expression on intratumoral APCs and tumor cells, and subsequently achieve targeted enrichment in tumors. Moreover, the efficacy of NDCs is biased toward dependence on host expression of PD-L1. CONCLUSIONS: The novel PD-L1/TLR7 dual-targeting NDC exhibited potent efficacy against heterogeneous tumors through orchestrating innate and adaptive immunity, which could act as a promising strategy to improve ICB therapy and shows prospects for clinical development.