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.

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.

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.

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.

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.

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.

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.

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.

Structural insights into the activation of human calcium-sensing receptor.

Human calcium-sensing receptor (CaSR) is a G-protein-coupled receptor that maintains Ca2+ homeostasis in serum. Here, we present the cryo-electron microscopy structures of the CaSR in the inactive and agonist+PAM bound states. Complemented with previously reported structures of CaSR, we show that in addition to the full inactive and active states, there are multiple intermediate states during the activation of CaSR. We used a negative allosteric nanobody to stabilize the CaSR in the fully inactive state and found a new binding site for Ca2+ ion that acts as a composite agonist with L-amino acid to stabilize the closure of active Venus flytraps. Our data show that agonist binding leads to compaction of the dimer, proximity of the cysteine-rich domains, large-scale transitions of seven-transmembrane domains, and inter- and intrasubunit conformational changes of seven-transmembrane domains to accommodate downstream transducers. Our results reveal the structural basis for activation mechanisms of CaSR and clarify the mode of action of Ca2+ ions and L-amino acid leading to the activation of the receptor.

Suppressive role of microRNA-130b-3p in ferroptosis in melanoma cells correlates with DKK1 inhibition and Nrf2-HO-1 pathway activation.

Cell death pathways related to ferroptosis are implicated in the progression of melanoma. Emerging data reporting the upregulation of microRNA (miR)-130b-3p in melanoma indicate the potential implication of miR-130b-3p in this malignancy. Herein, we aimed to identify whether and how miR-130b-3p regulated ferroptosis in melanoma cells. Melanoma cells (A375, G-361) were treated with erastin or RSL3 to mimic ferroptosis in vitro. Viability, lipid peroxidation level and ferrous ion content in melanoma cells were then assessed in response to manipulation of miR-130b-3p expression. Luciferase assay was conducted to determine the binding of miR-130b-3p to Dickkopf1 (DKK1). Western blot assay was conducted to determine the expression of molecules related to nuclear factor-erythroid 2 p45-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) pathway. The results indicated that miR-130b-3p exerted an inhibitory role in erastin or RSL3-induced ferroptosis, evidenced by reductions in lipid peroxidation and ferrous ion content. By suppressing the expression of target gene DKK1, miR-130b-3p activated the Nrf2/HO-1 pathway, whereby repressing ferroptosis. miR-130b-3p blocked the antitumor activity of erastin. Further, in vitro findings were reproduced in an in vivo murine model. Together, these data suggest the potential of miR-130b-3p to inhibit ferroptosis in melanoma cells and the mechanism was related to DKK1-mediated Nrf2/HO-1 pathway.

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.

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.

miR-21 promotes growth, invasion and migration of lung cancer cells by AKT/P-AKT/cleaved-caspase 3/MMP-2/MMP-9 signaling pathway.

OBJECTIVE: This study aimed to demonstrate the effects of miR-21 on the growth, migration, and invasion of lung cancer cells A549 in vitro and the possible mechanism. METHODS: In vitro cell migration and invasion potential were determined by Transwell chamber assays. FACS was used to assess the effect of miR-21 on A549 cell cycle and apoptosis. 4-6 week-old female mice were utilized to establish a lung cancer model. The pathologic biopsy was processed by H&E staining. The expression of the proteins PTEN, RECK and Caspase 3 were detected through immunohistochemy and tumor cell apoptosis was measured by TUNEL. RESULTS: Transwell chamber assays showed that the cells going through the membrane increased significantly compared to the negative control (P<0.05). The tumor volume resulting from miR-21 mimics was significantly greater than in normal mice. Serum ELISA showed that the protein expression levels of MMP-2 and MMP-9 in miR-21 overexpression group were increased significantly. In addition, H&E staining results showed that in miR-21 overexpression tissue, invasion is more severe and immunohistochemical results proved that the miR-21 overexpression group had high expression of Caspase 3 protein but the expression of PTEN and RECK were decreased. TUNEL experiments show that increased the expression of miR-21 can inhibit the apoptosis of tumor cells. CONCLUSION: MicroRNA-21 promotes the proliferation of lung cancer cells and inhibits the apoptosis of lung cancer cells by the AKT/P-AKT/cleaved-caspase 3/MMP-2/MMP-9 signaling pathway.

Overexpressed VEPH1 inhibits epithelial-mesenchymal transition, invasion, and migration of human cutaneous melanoma cells through inactivating the TGF-ß signaling pathway.

Malignant melanoma has a profound influence on populations around the world, with the underlying mechanisms controlling this disease yet to be fully identified. Hence, the current study aimed to investigate effects associated with VEPH1 on epithelial-mesenchymal transition (EMT), proliferation, invasion, migration and the apoptosis of human cutaneous melanoma (CM) cells through the TGF-ß signaling pathway. Microarray-based gene analysis was initially performed to screen the CM-related differentially expressed genes. The expression of VEPH1, TGF-ß signaling pathway- and EMT-related genes in CM tissues and cell lines was subsequently evaluated. Gain-of- and loss-of-function experiments were conducted to examine the effects of VEPH1 and the TGF-ß signaling pathway on the expression of EMT-related genes, cell proliferation, migration, invasion, cell cycle and apoptosis in vitro. Finally, tumor formation in nude mice was conducted. VEPH1 was lowly expressed and regulated the progression of CM with involvement in the TGF-ß signaling pathway. Human CM tissues were noted to activate the TGF-ß signaling pathway and EMT. A375 cells treated with overexpressed VEPH1 plasmids or/and TGF-ß signaling pathway inhibitor SB-431542 displayed diminished TGF-ß, SMAD4, Vimentin and N-cadherin expression while the expression of E-cadherin was elevated, accompanied by decreased cell proliferation, migration, invasion, inhibited cell cycle entry. However, si-VEPH1 or TGF-ß signaling pathway activator contributed to reverse results. Taken together, the key findings of the current study present evidence suggesting that VEPH1 protects against human CM by inhibiting the activation of the TGF-ß signaling pathway, highlighting its potential as a target for the prognosis and diagnosis of CM.

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.

Ozone therapy ameliorates inflammation and endometrial injury in rats with pelvic inflammatory disease.

As a common cause of infertility, pelvic inflammatory disease (PID) is characterized by chronic pain, ectopic pregnancy as well as inflammation and infection of the female upper genital tract. Ozone water, also known as O3, has been previously reported to be a distinctly effective agent in treating inflammation. During the present study, we asserted the hypothesis that O3 could be applied by pelvic inflammation and works to regulate the expression of inflammatory factors including interleukin-6 (IL-6), IL-2 and tumor necrosis factor-α (TNF-α). In an attempt to evaluate the effect of O3 on PID, an acute PID rat model was subsequently established. O3 at concentrations of 45 µg/mL and 60 µg/mL in addition to levofloxacin (LVLX) was injected respectively into the PID rats in a bid to alter the contents of inflammatory factors and immunologic markers. Hematoxylin-eosin (HE) staining was applied to analyze endometrial inflammation. Reductions to the contents of IL-6 and TNF-α were recorded, while that of IL-2, IgA, IgG, IgM, C3 and C4, and E rosette formation rate and transformation rate of T lymphocytes exhibited notably elevated levels after the PID rats had been injected with 45 µg/mL O3, 60 µg/mL O3 or LVLX. The pathological condition of the endometrium in rats with PID was alleviated among the PID rats after injected with the 45 µg/mL O3, 60 µg/mL O3 or LVLX. Taken together, the key findings of the current study present evidence demonstrating that the administration of O3 to the pelvic cavity ameliorated the PID conditions among rat models via inhibition of the necrosis of the endometrial epithelial cells as well as alleviated the inflammatory reactions, highlighting a potential novel PID treatment target.

[Application of early continuous renal replacement therapy in the bundle treatment of severe pneumonia].

OBJECTIVE: To explore the role of early continuous renal replacement therapy (CRRT) in bundle treatment of severe pneumonia. METHODS: Seventy severe pneumonia patients admitted to respiratory intensive care unit (RICU) of Second Affiliated Hospital of Xuzhou Medical University from January 2013 to June 2017 were enrolled. The patients were randomly divided into routine treatment group and CRRT treatment group, with 35 patients in each group. All patients were treated with bundle therapy, and those in CRRT treatment group was treated with CRRT daily on the basis of conventional bundle therapy. The mode was continuous veno-venous hemofiltration (CVVH), lasting 8-24 hours at a time. On the day of admission and the treatment of 1, 3, 5, 7 days, the fasting venous blood was collected. Inflammatory stress and immune parameters including procalcitonin (PCT), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), and CD4+ cells were determined by enzyme linked immunosorbent assay (ELISA) or flow cytometer. The incidence of acute respiratory distress syndrome (ARDS) and mortality within 7 days were observed in the two groups. RESULTS: Seventy patients with severe pneumonia were enrolled. There was no significant difference in the serum PCT, IL-6, TNF-α or CD4+ before treatment between the two groups. With the prolongation of treatment time, the PCT, IL-6 and TNF-α levels were gradually decreased, and the CD4+ was gradually increased. Compared with routine treatment group, PCT and TNF-α in CRRT treatment group were significantly decreased since the 3rd day [PCT (µg/L): 3.11±1.28 vs. 3.76±1.42, TNF-α (ng/L): 98.61±11.58 vs. 119.47±12.38], and CD4+ was significantly increased (0.39±0.03 vs. 0.35±0.03, all P < 0.05). The changes in IL-6 of CRRT treatment group was delayed, the statistical significance was found since the 5th day as compared with routine treatment group (ng/L: 35.43±12.39 vs. 52.86±10.78, P < 0.05). Compared with routine treatment group, the incidence of ARDS [11.43% (4/35) vs. 31.43% (11/35)] and mortality [0% (0/35) vs. 11.43% (4/35)] within 7 days in CRRT treatment group were significantly lowered (both P < 0.05). CONCLUSIONS: CRRT in the early stage of bundle therapy for severe pneumonia is not only suggested to remove inflammatory mediators and improve immune function, but an opportunity and effective way to reduce complications and delay rapid progression of severe pneumonia. It provides the opportunity and condition for comprehensive treatment.

Hollow Mesoporous Silica@Metal-Organic Framework and Applications for pH-Responsive Drug Delivery.

Metal--organic frameworks (MOFs), a new type of porous crystalline material, hold great potential in biomedical applications, such as drug delivery. However, the efficacy of drug delivery is limited by low drug loading. In this work, we synthesized hollow mesoporous silica (HMS)@MOF capsules that can be used as a pH-responsive drug delivery system for the anticancer drug doxorubicin (DOX). DOX is loaded into the inner cavity of HMS. Zeolitic imidazolate framework-8 (ZIF-8) nanoparticles are then coated on the outer surface of the DOX-loaded HMS. The obtained material is a capsule (denoted as DOX/HMS@ZIF), in which DOX is encapsulated. The DOX/HMS@ZIF can be used as an efficient pH-responsive drug delivery system. DOX is not released under physiological conditions (pH 7.4), but is released at low pH (4-6) from DOX/HMS@ZIF. The DOX/HMS@ZIF capsule shows much higher cytotoxicity than free DOX and alters the delivery pathway for DOX in cancer cells, while the drug-free HMS@ZIF shows excellent biocompatibility. This opens new opportunities to construct a safe and efficient delivery system for targeted molecules using pH-responsive release for a wide range of applications.