An additional medium renewal of D4 embryo culture improves the concordance of noninvasive chromosome screening with trophectoderm biopsy.

Our research question was to evaluate the chromosome concordance of trophectoderm (TE) biopsy with noninvasive chromosome screening (NICS) using embryo culture medium renewed twice on Day 3 (D3) and Day 4 (D4). In this study, we evaluated 64 cycles with 223 biopsied blastocysts. These were categorized into two groups based on replacing embryo culture medium on D3 (control group) or on D3 and D4 (experimental group). The fundamental characteristics and main outcomes were compared. The concordance rates of NICS results with TE biopsy were determined according to next generation sequencing results. In total, 103 experimental and 120 control embryo cultures were collected, and the euploid status was analyzed using NICS technology. The overall concordance rates with TE biopsy of the experimental and control groups were 0.86 and 0.75, respectively. Statistically significant difference was found between the two groups. An additional medium renewal of the D4 embryo culture can improve the concordance of NICS with TE biopsy.

CLDN1 regulates trophoblast apoptosis and proliferation in preeclampsia.

Preeclampsia is a gestational hypertensive disease; however, preeclampsia remains poorly understood. Bioinformatics analysis was applied to find novel genes involved in the pathogenesis of preeclampsia and identified CLDN1 as one of the most differentially expressed genes when comparing patients with preeclampsia and healthy controls. The results of the qRT-PCR, Western blotting and immunohistochemistry experiments demonstrated that CLDN1 was significantly downregulated in the chorionic villi in samples from patients with preeclampsia. Furthermore, knockdown of CLDN1 in HTR-8/SVneo cells resulted in the inhibition of proliferation and induction of apoptosis, and overexpression of CLDN1 reversed these effects. In addition, RNA-seq assays demonstrated that the gene BIRC3 is potentially downstream of CLDN1 and is involved in the regulation of apoptosis. Knockdown of CLDN1 confirmed that the expression level of BIRC3 was obviously decreased and was associated with a significant increase in cleaved PARP. Interestingly, the apoptotic effect in CLDN1 knockdown cells was rescued after BIRC3 overexpression. Overall, these results indicate that a decrease in CLDN1 inhibits BIRC3 expression and increases cleaved PARP levels thus participating in the pathogenesis of preeclampsia.

Expert Consensus on Polymyxin Antimicrobial Susceptibility Testing and Clinical Interpretation.

The polymyxins are important antimicrobial agents against antibiotic-resistant gram-negative bacilli. In 2020, the Clinical and Laboratory Standards Institute modified the clinical breakpoints for polymyxin susceptibility test by eliminating the "susceptible" interpretive category, only reporting intermediate (≤2 mg/L) and resistant (≥4 mg/L). However, the European Committee on Antimicrobial Susceptibility Testing recommended the use of clinical breakpoints of ≤2 mg/L as susceptible and >2 mg/L as resistant. The first-line laboratorians and clinicians in China have been perplexed by the inconsistence of international polymyxin clinical breakpoints and discouraged by the difficulty of conducting polymyxin susceptibility testing. Therefore, it is urgently needed to make it clear for the laboratorians in China to know how to accurately carry out polymyxin susceptibility testing and standardize the interpretation of susceptibility testing results. To this end, the experts from relevant fields were convened to formulate this consensus statement on the testing and clinical interpretation of polymyxin susceptibility. Relevant recommendations are proposed accordingly for laboratorians and clinicians to streamline their daily work.

PDIA3 regulates trophoblast apoptosis and proliferation in preeclampsia via the MDM2/p53 pathway.

Protein disulfide isomerase 3 (PDIA3) is a chaperone protein that modulates the folding of newly synthesized glycoproteins, has isomerase and redox activity, and has been implicated in the pathogenesis of many diseases. However, the role of PDIA3 in pregnancy-associated diseases remains largely unknown. Our present study reveals a key role for PDIA3 in the biology of placental trophoblasts from women with preeclampsia (PE). Immunohistochemistry and Western blot analysis revealed that PDIA3 expression was decreased in villous trophoblasts from women with PE compared to normotensive pregnancies. Further, using a Cell Counting Kit-8 assay, flow cytometry, and 5-ethynyl-2'-deoxyuridine (EdU) staining, we found that siRNA-mediated PDIA3 knockdown significantly promoted apoptosis and inhibited proliferation in the HTR8/SVneo cell line, while overexpression of PDIA3 reversed these effects. Furthermore, RNA sequencing and Western blot analysis demonstrated that knockdown of PDIA3 inhibited MDM2 protein expression in HTR8 cells, concurrent with marked elevation of p53 and p21 expression. Conversely, overexpression of PDIA3 had the opposite effects. Immunohistochemistry and Western blot further revealed that MDM2 protein expression was downregulated and p21 was increased in trophoblasts of women with PE compared to women with normotensive pregnancies. Our findings indicate that PDIA3 expression is decreased in the trophoblasts of women with PE, and decreased PDIA3 induces trophoblast apoptosis and represses trophoblast proliferation through regulating the MDM2/p53/p21 pathway.

Quantitative proteomic analysis reveals that Luks-PV exerts antitumor activity by regulating the key proteins and metabolic pathways in HepG2 cells.

Hepatocellular carcinoma (HCC) is a complicated and poor prognosis cancer, necessitating the development of a potential treatment strategy. In this study, we initially revealed that LukS-PV belonged to leukocidin family performs an anti-HCC action. Then, we used liquid chromatography-mass spectrometry (LC/MS) to compare protein expression profiles of the LukS-PV-treated human HCC cell lines HepG2 and the control cells. GO annotations and Kyoto Encyclopedia of Genes and Genomes pathway analysis were carried out of differential expression followed by protein-protein interactome, to explore the underlying cancer suppressor mechanisms of LukS-PV for human HCC. A total of 88 upregulated proteins and 46 downregulated proteins were identified. The top 10 proteins identified by the MCC method are FN1, APP, TIMP1, nucleobindin-1, GOLM1, APLP2, CYR61, CD63, ENG, and CD9. Our observation on protein expression indicated that LukS-PV produces a signature affecting central carbon metabolism in cancer, galactose metabolism, and fructose and mannose metabolism pathways. The results give a functional effects and molecular mechanism insight, following LukS-PV treatment.

[Chemical constituents from green walnut husks and their antitumor activity in vitro].

Fourteen chemical constituents, including 5-hydroxy-4-methoxy-1-tetralone(1), 4,8-dihydroxy-1-tetralone(2), 4,5-dihydroxy-α-tetralone(3), blumenol B(4), dehydrovomifoliol(5), megastigm-5-ene-3,9-diol(6), juglanin B(7), blumenol C(8), loliolide(9), oleracone B(10), syringarsinol(11), pinoresinol(12), methyl 4-hydroxy-3-methoxybenzoate(13), and isovanillic acid(14), were isolated from the dichloromethane fraction of 95% methanol extract of green walnut husks by silica gel and MCI column chromatography, and Pre-HPLC. Their structures were determined by spectroscopic methods, such as NMR, MS and so on. Among them, compounds 1, 4-6, 8-13 were isolated from the green walnut husks for the first time, and compounds 4-6, 8, 10, 12, 13 were isolated from the Juglans genus for the first time. All of isolates were detected their inhibitory activities against HeLa, HGC-27 and Ht-29 cell lines by the MTT assay. The result showed that compounds 2, 3, 7, 9 and 11 exhibited inhibitory activity against the tested cell line. The IC_(50) of 7 were 26.5, 9.0, 25.4 µmol·L~(-1), respectively.

The YY1-HOTAIR-MMP2 Signaling Axis Controls Trophoblast Invasion at the Maternal-Fetal Interface.

We aimed to determine the effect of YY1 expression on the expression profile of long noncoding RNAs (lncRNAs) in trophoblasts, and we studied the involvement of certain lncRNAs and YY1 in the pathogenesis of recurrent miscarriage (RM). RT2 lncRNA PCR arrays revealed that YY1 overexpression in trophoblasts significantly promoted the expression of the HOX transcript antisense RNA HOTAIR and demonstrated that HOTAIR expression was significantly lower in the RM trophoblasts than in control trophoblasts. Ectopic HOTAIR overexpression and knockdown experiments revealed that it was a novel target of YY1. Bioinformatics analysis identified two YY1-binding sites in the HOTAIR promoter region, and chromatin immunoprecipitation (ChIP) analysis verified that YY1 binds directly to its promoter region. Interestingly, HOTAIR overexpression enhanced trophoblast invasion in an ex vivo explant culture model, while its knockdown repressed these effects. Furthermore, liquid chromatography-tandem mass spectrometry (LC-MS/MS) label-free quantitative proteomics screening revealed that HOTAIR overexpression activated phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) signaling in trophoblasts. In an ex vivo explant culture model, HOTAIR overexpression effectively elevated matrix metalloproteinase 2 (MMP2) expression via the PI3K-AKT signaling pathway, enhancing trophoblast migration and invasion. These findings reveal a new regulatory pathway in which YY1 activates PI3K-AKT signaling via HOTAIR, promoting MMP2 expression, suggesting that HOTAIR is a potential therapeutic target for RM.

Four New Sesquiterpenoids from the Roots of Diarthron Tianschanica with Their Antineoplastic Activity.

Four new sesquiterpenoids, known as diarthronchas A⁻D (1⁻4), and one known daphnauranol B (5) were isolated from the methanol extract of the roots of Diarthron tianschanica. The compounds structures were determined on the basis of spectroscopic data. All of the isolated compounds were profiled for their antineoplastic activity.

LukS-PV-Regulated MicroRNA-125a-3p Promotes THP-1 Macrophages Differentiation and Apoptosis by Down-Regulating NF1 and Bcl-2.

BACKGROUND/AIMS: LukS-PV is a component of Panton-Valentine leukocidin (PVL). We have previously demonstrated that LukS-PV potently promoted differentiation and induced apoptosis in THP-1 cells. However, the precise mechanisms of these actions remain unknown. MicroRNAs (miRs) play important roles in cellular differentiation and apoptosis. This study aimed to investigate the role of miR-125a-3p in LukS-PV-regulated differentiation and apoptosis and its underlying mechanism in THP-1 cells. METHODS: MicroRNA profiling analyses were conducted to determine differential miRNA expression levels in THP-1 cells treated with LukS-PV. Cell differentiation and apoptosis were measured in THP-1 cells by gain-of-function and loss-of-function experiments. Bioinformatics analysis and luciferase reporter assays were used to confirm the targets of miR-125a-3p. The effects of the miR-125a-3p targets on cellular differentiation were determined by knocking them down. RESULTS: MiR-125a-3p was up-regulated after treating the human monocytic leukaemia cell line THP-1 with LukS-PV. In addition, miR-125a-3p positively regulated apoptosis and differentiation in THP-1 cells treated with LukS-PV. Concordantly, luciferase reporter assays confirmed that neurofibromatosis type 1 (NF1) and B-cell lymphoma 2 (Bcl-2) were direct target genes of miR-125a-3p. Moreover, NF1 knockdown in THP-1 cells significantly promoted differentiation in vitro. Finally, the extracellular signal-regulated kinase (ERK) pathway, a downstream target of NF1, was activated after NF1 knockdown. CONCLUSIONS: These findings confirm that miR-125a-3p is involved in LukS-PV-mediated cell differentiation and apoptosis in THP-1 cells.

The YY1/MMP2 axis promotes trophoblast invasion at the maternal-fetal interface.

YY1 is a sequence-specific DNA-binding transcription factor that has many important biological roles. However, its function in trophoblasts at the maternal-fetal interface remains to be elucidated. In this study, we used an mRNA microarray and reverse transcription qPCR and compared the YY1 mRNA expression level in trophoblasts between patients with recurrent miscarriage (RM) and healthy control subjects. Our results revealed that YY1 mRNA expression was significantly lower in the trophoblasts of the RM group compared with the healthy control group. Furthermore, immunofluorescence and immunohistochemical data showed that YY1 was highly expressed in human placental villi during early pregnancy, especially in cytotrophoblast cells and invasive extravillous trophoblasts, and it was expressed at a much lower level in the placental villi of term pregnancy. YY1 overexpression enhanced, and knockdown repressed, the invasion and proliferation of trophoblasts. Antibody array screening revealed that YY1 significantly promoted MMP2 expression in trophoblasts. Bioinformatics analysis identified three YY1-binding sites in the MMP2 promoter region, and chromatin immunoprecipitation analysis verified that YY1 binds directly to its promoter region. Importantly, inhibition of YY1 by siRNA clearly decreased trophoblast invasion in an ex vivo explant culture model. Overall, our findings revealed a new regulatory pathway of YY1/MMP2 in trophoblast cell invasion during early pregnancy and indicated that YY1 may be involved in the pathogenesis of RM.

Study on Spatial Distribution and Seasonal Variations of Trace Metal Contamination in Sediments from the Three Adjacent Areas of the Yellow River Using HR-ICP-MS.

Spatial distribution, seasonal variations and pollution conditions of 6 heavy metals (Cr, Pb, Cd, Cu, Zn and Ni) in sediments in three different seasons (wet, dry and normal) from Gansu province, Ningxia and Inner Mongolia Autonomous Regions of the Yellow River were studied using high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS), geo-accumulation index (Igeo), respectively. The results indicated that content of heavy metals in all three seasons of Inner Mongolia sediment samples were higher compared to Ningxia and Gansu sections, and except Cd, the other metals concentrations in wet season are relatively higher, and they are lower and approximated in dry and normal seasons. Igeo showed that high Igeo values of Cd, Cu and Cr were found in Inner Mongolia region in wet and dry seasons, and the low Igeo values of the three metals were obtained in Gansu region. For season, the Igeo values of all study metals were higher in wet season than dry season, and they were decreased in normal season except Cd, while ICdgeo were high in all sampling sites in normal season indicating the strong anthropogenic sources there. Igeo values of metals including Pb, Zn and Ni were below 0 in wet season of Gansu, in dry season of Gansu, Ningxia regions and in normal season of all study regions, respectively, indicating a safer level. The results of spatial distribution and Igeo of study heavy metals suggested that higher total metal concentrations and potential risks were found in Inner Mongolia reaches of the Yellow River, and Cr, Cd and Cu were being more significant with the strong anthropogenic sources among all study metals. Considering the three different seasons, except normal season, the higher metal concentrations and potential risks for metals in sediments were observed in wet season than dry season, since the higher pH values of the sedimentary samples in wet season can help to promote adsorption and precipitation of heavy metal in sediments, meanwhile, the higher total organic carbon (TOC) values in sediments in wet season were also beneficial to the adsorption of heavy metals since the affinity between TOC and metal cations. Besides, the coast sediments with variety pollutants were brought into the river by rainfall in summer, which may also contribute to the rapid enrichment of heavy metals in sediments in wet season than dry season. The correlation among heavy metals and other variables and potential sources for heavy metals were evaluated by correlation coefficient analysis (CCA). The results of CCA revealed that Pb, Cu, Zn and Ni showed a significant correlation with each other, and they would be easily influenced by pH and TOC. Furthermore, Cr showed a correlation with Cd and TOC and reflected a strong anthropogenic source. The present work could provide new information on the metals behavior in surface sediments and reflect the sediment geochemistry in the regions in the three seasons, whilst the study of element interrelationships, including pH and TOC gave the available information on their possible origins.

Studied on Distribution and Heavy Metal Pollution Index of Heavy Metals in Water from Upper Reaches of the Yellow River, China.

In order to understand the spatial distribution and evaluate the pollution degree of heavy metals (As, Cd, Cr, Co, Cu, Mn, Ni, Pb, and Zn) in upper reaches of the Yellow River, surface water samples were collected from 12 selected sites during two field surveys in April 2014 (drought season) and October 2014 (normal season). The concentrations of heavy metals were determined using inductively coupled plasma mass spectrometry (ICP-MS) for spatial variation and heavy metal pollution index. The average concentrations of the metals in the drought season and normal season decreased respectively in the order: Cr (18.56 µg·L-1)>As (2.95 µg·L-1)>Ni (1.87 µg·L-1)>Mn (1.20 µg·L-1)>Cu (1.12 µg·L-1)>Zn (0.59 µg·L-1)>Pb (0.08 µg·L-1)>Cd (0.01 µg·L-1); Mn (596.89 µg·L-1)>Zn (52.46 µg·L-1)>Cu (36.27 µg·L-1)>Ni (25.11 µg·L-1)>Cr (23.19 µg·L-1)>Pb (19.51 µg·L-1)>As (7.30 µg·L-1)>Cd (0.37 µg·L-1). The results were compared with national and international water quality guidelines, as well as literature values reported for the same river in flood season. To assess the composite influence of all the considered metals on the overall quality of the water, heavy metal pollution indices were calculated. The Heavy Metal Pollution Index (HPI) of the river calculated for the individual locations showed ranging from 6.46 to 11.95 in drought season, 4.53 to 210.53 in normal season, respectively. Both the distribution of metals and HPI of two seasons revealed obvious seasonal variation. The results showed that the degree of heavy metal contaminant was not very high and had seasonal variation; both the concentrations of metals and HPIs indicated the pollution level of the normal season was higher than it of the drought season. The results of this paper provided reliable experimental data and theoretical basis for the relevant departments to make further policy decision.

SRC2-1 is required in PcINF1-induced pepper immunity by acting as an interacting partner of PcINF1.

Elicitins are elicitors that can trigger hypersensitive cell death in most Nicotiana spp., but their underlying molecular mechanism is not well understood. The gene Phytophthora capsici INF1 (PcINF1) coding for an elicitin from P. capsici was characterized in this study. Transient overexpression of PcINF1 triggered cell death in pepper (Capsicum annuum L.) and was accompanied by upregulation of the hypersensitive response marker, Hypersensitive Induced Reaction gene 1 (HIR1), and the pathogenesis-related genes SAR82, DEF1, BPR1, and PO2. A putative PcINF1-interacting protein, SRC2-1, was isolated from a pepper cDNA library by yeast two-hybrid screening and was observed to target the plasma membrane. The interaction between PcINF1 and SRC2-1 was confirmed by bimolecular fluorescence complementation and co-immunoprecipitation. Simultaneous transient overexpression of SRC2-1 and PcINF1 in pepper plants triggered intensive cell death, whereas silencing of SRC2-1 by virus-induced gene silencing blocked the cell death induction of PcINF1 and increased the susceptibility of pepper plants to P. capsici infection. Additionally, membrane targeting of the PcINF1-SRC2-1 complex was required for cell death induction. The C2 domain of SRC2-1 was crucial for SRC2-1 plasma membrane targeting and the PcINF1-SRC2-1 interaction. These results suggest that SRC2-1 interacts with PcINF1 and is required in PcINF1-induced pepper immunity.

[Study on Speciation Analysis and Ecological Risk Assessment of Heavy Metals in Surface Sediments in Gansu, Ningxia and Inner Mongolia Sections of the Yellow River in Wet Season with HR-ICP-MS].

In order to continuously study the contents, pollution condition and potential ecological risk of heavy metals in surface sediments in Gansu, Ningxia and Inner Mongolia sections of the Yellow River in wet seasons in different years, the speciation analysis of 9 kinds of heavy metals including Cd, Pb, Cr, Ni, Cu, V, Co, Zn and Mn, pollution condition and potential ecological risk of heavy metals in surface sediments from 10 sampling sites like Baotoufuqiao (S2), Shizuishantaolezhen (S6) and Wujinxia (S9) in Gansu, Ningxia and Inner Mongolia sections of the Yellow River in 2012 wet season were studied with BCR sequential extraction and high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS) based on our previous works. The results implied that the order of heavy metals average contents in the 10 sediment samples were the same: Mn>V> Zn>Cr>Cu>Ni>Pb>Co>Cd. In the sediments, heavy metals mainly existed in the form of residual fraction, which indicated that the bioavailability or environmental impact was low. Results of geo-accumulation indices (Igeo) showed that Igeo(CD), was the largest among the heavy metals with the strongest pollution, while IGEO(Mn)was the smallest. Enrichment factor (EF) indicated that only Cd and Cu were enriched at some sampling sites. In S5, because EFcd reached 4. 69, Cd was affected by human activities obviously and the result was consistent with I. Potential ecological risk index (RI) implied that the RI values in S1, S2 and S5 were between 150 and 300, which belonged to moderate polluting degree, while others were less than 150, belonging to light pollution degree. The results of this paper could not only provide reliable experimental data and theoretical basis for the relevant departments, but also supply the technical support for constructing mathematics model of sediments-pollutants transport, systematically researching the migration and transformation rule of persistent toxic substances and environmental assessment in these reaches.

[Research on the Content Characteristics and Pollution Evaluation of Heavy Metals in Filtered Water and Suspended Particles from Gansu, Ningxia and Inner Mongolia Sections of the Yellow River in Wet Season Using HR-ICP-MS].

The content characteristics, pollution evaluation and source identification of 6 heavy Metals (Cd, Pb, Cr, As, Cu and Zn) in filtered water and 9 heavy Metals (Cd, Pb, Cr, Ni, Cu, V, Co, Zn and Mn) in suspended particles from 10 sampling sites such as Zhaojunfuqiao (S1) and Baotoufuqiao (S2), etc. from Gansu, Ningxia and Inner Mongolia sections of the Yellow River in 2012 Wet Season were studied to understand the condition of the heavy metal pollution in Gansu, Ningxia and Inner Mongolia Sections of the Yellow River by using high resolution inductively coupled plasma-mass spectrometry (HR-ICP-MS). Multivariate geochemical approaches and statistical analysis were also exploited for assessing the level of heavy metals in filtered water and suspended particles from studied area. The results showed that in filtering water, only the concentrations of Cr exceeded the standard value of Environmental Quality Standard for Surface Water (GB3838-2002) and were the highest (74.8-94.7 µg x L(-1)) among all elements in 10 sampling sites; Single factor pollution index (I(i)) results suggested that the water quality in all sampling sites were contaminated by both Cr and total nitrogen (TN), with the exception of TN in Baotoufuqiao (S2); Integrated Nemerow pollution index (I) indicated that the I values in all sampling sites were between 1-2 (light pollution), which implied that the water quality in Gansu, Ningxia and Inner Mongolia sections, especially downstream sections (S1-S6) of the Yellow River wasn't an ideal source for drinking and using in aquaculture any more. In suspended particles, concentrations of heavy metals were relatively higher than their soil background values in 10 sampling sites, except Ni in S10 (34.7 µg x L(-1)). Index of geo-accumulation (I(geo)) indicated that the I(geo) values of Pb, Cr, Ni, Cu, V, Co, Zn and Mn in all sampling sites were less than 1 (unpolluted or unpolluted-moderately polluted), respectively, while I(geo)Cd were the highest in 10 sampling sites among all heavy metals and with the moderately to strong contamination in Zhaojunfuqiao (S1), Baotoufuqiao (S2), Wuhai (S5) and Dongdagouruhuanghekou (S8). The results of this paper would help to supply reliable experimental data for researching of distribution, migration and effective protection of heavy metals in study area.

Diffusion simulation and risk assessment model establishment of chlorine gas leakage based on terrain conditions.

This study researches the impact of terrain factors on chlorine gas diffusion processes based on SLAB model. Simulating the law of wind speed changing with altitude by calculating the real-time speed with vertical height combing actual terrain data, and integrating the influence of terrain on wind speed by using Reynolds Average Navier-Stokes (RANS) algorithm, K-turbulence model, and standard wall functions, then plotting the gas diffusion range in the map with terrain data according to the Gaussian-Cruger projection algorithm and dividing the hazardous areas according to the public exposure guidelines (PEG). The accidental chlorine gas releases near Lishan Mountain, Xi'an City, were simulated by the improved SLAB model. The results show that there are obvious differences analyzing contrastively the endpoint distance and area of chlorine gas dispersion under real terrain condition and ideal condition at different times; it can be found that the endpoint distance of the real terrain conditions is 1.34 km shorter than that of the ideal conditions at 300 s with terrain factors, and also the thermal area is 3,768,026m2 less than that of the ideal conditions. In addition, it can predict the specific number of casualties within different levels of harm at 2 min after chlorine gas dispersion, and casualties are constantly changing over time. The fusion of terrain factors can be used to optimize the SLAB model, which is expected to provide an important reference for effective rescue.

Mechanism of Procyanidin B2 in the Treatment of Chronic Myeloid Leukemia Based on Integrating Network Pharmacology and Molecular Docking.

OBJECTIVE: To study the pharmacological mechanism of procyanidin B2 (PCB2) on chronic myeloid leukemia (CML) by integrating network pharmacological methods systematically. METHODS: Firstly, the potential target genes of PCB2 were predicted by the pharmacological database and analysis platform (TCMSP and Pharmmapper). Meanwhile, the relevant target genes of CML were collected from GeneCards and DisGene. Pooled data were collected to screen for common target genes. Furthermore, the above intersection genes were imported into the String website to construct a protein-protein interaction (PPI) network, and the Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were further analyzed. Besides, molecular docking was performed to verify the possible binding conformation between PCB2 and candidate targets. Finally, MTT and RT-PCR experiments of K562 cells were performed to verify the above results of network pharmacology. RESULTS: A total of 229 PCB2 target genes were retrieved, among which 186 target genes had interaction with CML. The pharmacological effects of PCB2 on CML were related to some important oncogenes and signaling pathways. The top ten core targets predicted by Network Analysis were as follows: AKT1, EGFR, ESR1, CASP3, SRC, VEGFA, HIF1A, ERBB2, MTOR, and IGF1. Molecular docking studies confirmed that hydrogen bonding was the main interaction force of PCB2 binding targets. According to the molecular docking score, the following three target proteins were most likely to bind to PCB2: VEGFA (-5.5 kcal/mol), SRC (-5.1 kcal/mol), and EGFR (-4.6 kcal/mol). After treatment of PCB2 for 24h, mRNA expression levels of VEGFA and HIF1A decreased significantly in K562 cells. CONCLUSION: Through integrating network pharmacology combined with molecular docking, the study revealed the potential mechanism of PCB2 anti-chronic myeloid leukemia.

Mechanism of human chorionic gonadotropin in endometrial receptivity via the miR-126-3p/PI3K/Akt/eNOS axis.

Human chorionic gonadotropin (hCG) might affect endometrial receptivity, exerting integral roles in embryo implantation. This study explored the action of hCG in endometrial receptivity via the miR-126-3p/PIK3R2/PI3K/Akt/eNOS axis. The embryo implantation dysfunction (EID) mouse models were established by administrating mifepristone and human endometrial epithelial cells (EECs) were used for in vivo experiments, both followed by hCG treatment. Expression level of CD105 and protein levels of cadherin CD144 and CD146 in mice were determined by immunohistochemistry and Western blot. The levels of miR-126-3p and PIK3R2 mRNA and PIK3R2, p-PI3K p85 α, PI3K p110 α, p-Akt, Akt, p-eNOS, and eNOS protein levels were measured. Cell proliferation was evaluated by CCK-8 and EdU assays. The binding sites of miR-126-3p and PIK3R2 were predicted and verified. hCG-treated EECs were further transfected with miR-126-inhibitor for functional rescue experiments. hCG ameliorated endometrial receptivity in EID mice. Moreover, hCG promoted miR-126-3p and suppressed PIK3R2 in EID mice and EECs. miR-126-3p targeted PIK3R2. EEC proliferation was enhanced after hCG treatment but inhibited by miR-126-3p downregulation. Both in vivo and in vitro experiments validated that hCG activated the PI3K/Akt/eNOS pathway through the miR-126-3p/PIK3R2 axis. Collectively, hCG improves endometrial receptivity by activating the PI3K/Akt/eNOS pathway via regulating miR-126-3p/PIK3R2.

18ß-glycyrrhetinic acid inhibits proliferation of gastric cancer cells through regulating the miR-345-5p/TGM2 signaling pathway.

BACKGROUND: Gastric cancer (GC) is a common gastrointestinal malignancy worldwide. Based on cancer-related mortality, the current prevention and treatment strategies for GC still show poor clinical results. Therefore, it is important to find effective drug treatment targets. AIM: To explore the molecular mechanism of 18ß-glycyrrhetinic acid (18ß-GRA) regulating the miR-345-5p/TGM2 signaling pathway to inhibit the proliferation of GC cells. METHODS: CCK-8 assay was used to determine the effect of 18ß-GRA on the survival rate of GES-1 cells and AGS and HGC-27 cells. Cell cycle and apoptosis were detected by flow cytometry, cell migration was detected by a wound healing assay, the effect of 18ß-GRA on subcutaneous tumor growth in BALB/c nude mice was investigated, and the cell autophagy level was determined by MDC staining. TMT proteomic analysis was used to detect the differentially expressed autophagy-related proteins in GC cells after 18ß-GRA intervention, and then the protein-protein interaction was predicted using STRING (https://string-db.org/). MicroRNAs (miRNAs) transcriptome analysis was used to detect the miRNA differential expression profile, and use miRBase (https://www.mirbase/) and TargetScan (https://www.targetscan.org/) to predict the miRNA and complementary binding sites. Quantitative real-time polymerase chain reaction was used to detect the expression level of miRNA in 18ß-GRA treated cells, and western blot was used to detect the expression of autophagy related proteins. Finally, the effect of miR-345-5p on GC cells was verified by mir-345-5p overexpression. RESULTS: 18ß-GRA could inhibit GC cells viability, promote cell apoptosis, block cell cycle, reduce cell wound healing ability, and inhibit the GC cells growth in vivo. MDC staining results showed that 18ß-GRA could promote autophagy in GC cells. By TMT proteomic analysis and miRNAs transcriptome analysis, it was concluded that 18ß-GRA could down-regulate TGM2 expression and up-regulate miR-345-5p expression in GC cells. Subsequently, we verified that TGM2 is the target of miR-345-5p, and that overexpression of miR-345-5p significantly inhibited the protein expression level of TGM2. Western blot showed that the expression of autophagy-related proteins of TGM2 and p62 was significantly reduced, and LC3II, ULK1 and AMPK expression was significantly increased in GC cells treated with 18ß-GRA. Overexpression of miR-345-5p not only inhibited the expression of TGM2, but also inhibited the proliferation of GC cells by promoting cell apoptosis and arresting cell cycle. CONCLUSION: 18ß-GRA inhibits the proliferation of GC cells and promotes autophagy by regulating the miR-345-5p/TGM2 signaling pathway.