Rapamycin circumvents anti PD-1 therapy resistance in colorectal cancer by reducing PD-L1 expression and optimizing the tumor microenvironment.

The unresectable or postoperative recurrence of advanced metastatic colorectal cancer (CRC) is the difficulty of its clinical management, and pharmacological therapy is the main source of benefit. Immune checkpoint inhibitors are therapeutic options but are effective in approximately 5 % of patients with deficient mismatch repair (MMR)/microsatellite instability CRC and are ineffective in patients with MMR-proficient (pMMR)/microsatellite stable (MSS) CRCs, which may be associated with the tumor microenvironment (TME). Here, we propose a new combination strategy and evaluate the efficacy of rapamycin (Rapa) combined with anti-PD-1 (αPD-1) in CT26 tumor-bearing mice, azoxymethane (AOM)/dextran sodium sulfate (DSS) inflammation-associated CRC mice, CT26-Luc tumor-bearing mice with postoperative recurrence, and CT26 liver metastasis mice. The results revealed that Rapa improved the therapeutic effect of αPD-1 and effectively inhibited colorectal carcinogenesis, postoperative recurrence, and liver metastasis. Mechanistically, Rapa improved the anticancer effect of αPD-1, associated with Rapa reprograming of the immunosuppressive TME. Rapa effectively depleted α-SMA+ cancer-associated fibroblasts and degraded collagen in the tumor tissue, increasing T lymphocyte infiltration into the tumor tissue. Rapa induced the downregulation of programed cell death 1 ligand 1 (PD-L1) protein and transcript levels in CT26 cells, which may be associated with the inhibition of the mTOR/P70S6K signaling axis. Furthermore, co-culture of tumor cells and CD8+ T lymphocytes demonstrated that Rapa-induced PD-L1 downregulation in tumor cells increased spleen-derived CD8+ T lymphocyte activation. Therefore, Rapa improves the anti-tumor effect of αPD-1 in CRCs, providing new ideas for its use to improve combinatorial strategies for anti-PD-1 immunotherapy.

Nanoliposomes Encapsulated Rapamycin/Resveratrol to Induce Apoptosis and Ferroptosis for Enhanced Colorectal Cancer Therapy.

OBJECTIVES: Monotherapy is often ineffective for treating colorectal cancer. In this study, we developed PEG-modified liposomes loaded with rapamycin (Rapa) and resveratrol (Res) (Rapa/Res liposomes, or RRL) to investigate their therapeutic potential in colorectal cancer. METHODS: RRL were constructed using the reversed-phase evaporation method. We assessed the cytotoxicity, apoptosis, and ferroptotic effects of RRL on colorectal cancer HCT116 cells. The anti-tumor efficacy of RRL was evaluated in HCT116 xenograft mice. RESULTS: RRL had a particle size of 86.67 ± 1.10 nm and a zeta potential of -33.13 ± 0.49 mV. The coloaded formulation demonstrated satisfactory performance both in vitro and in vivo, resulting in increased cytotoxicity to HCT116 cells and significant suppression of HCT116 xenografts tumor growth. Mechanically, RRL significantly increased the apoptosis rate of HCT116 cells, induced ROS accumulation in tumor cells, and effectively downregulated the expression of the ferroptosis-associated proteins GPX4 and SLC7A11, demonstrating its superior efficacy compared to that of Rapa liposomes (Rapa/Lps) or Res liposomes (Res/Lps) alone. CONCLUSION: Coloading Rapa and Res into liposomes to promote apoptosis and ferroptosis in tumor cells represents a promising strategy for the treatment of colorectal cancer.

Stromal and tumor immune microenvironment reprogramming through multifunctional cisplatin-based liposomes boosts the efficacy of anti-PD-1 immunotherapy in pancreatic cancer.

The dense stromal barrier in pancreatic cancer tissues blocks intratumoral delivery and distribution of chemotherapeutics and therapeutic antibodies, causing poor chemoimmunotherapy responses. We designed a multi-targeted pH-sensitive liposome which encapsulates cisplatin (Pt) in its water core (denoted as ATF@Pt Lps) and shows high affinity for uPAR receptors in pancreatic cancer cells, tumor-associated macrophages, and cancer-associated fibroblasts. Systemic administration of ATF@Pt Lps enabled overcoming the central stromal cellular barrier and effective drug delivery into tumor cells, resulting in a strong therapeutic response in a Panc02 cell derived transplanted tumor mouse model. More importantly, ATF@Pt Lps degradation of collagen contributes to the infiltration of CD8+ T cells into tumors as well as an enhanced accumulation of anti PD-1 monoclonal antibodies. Furthermore, the killing of tumor cells by Pt also leads to the release of tumor antigens, which promote the proliferation of immune cells, especially CD83+ cells, Th1 CD4+ cells, and CD8+ cytotoxic T cells, that converted an immunoscore "cold" pancreatic cancer into a pro-immune "hot" tumor. A further combination with an immune checkpoint agent, anti PD-1 antibodies that inhibit PD-1, can enhance tumor specific cytotoxic T cell response. Accordingly, ATF@Pt Lps displays multi-targeting, controlled drug release, stromal disruption, enhanced penetration, killing of cancer cells, modification of the immunosuppressive microenvironment, and enhancement of immunity. This study provides important mechanistic information for the further development of a combination of ATF@Pt Lps and anti PD-1 antibodies for the effective treatment of pancreatic cancer.

Multi-targeting liposomal codelivery of cisplatin and rapamycin inhibits pancreatic cancer growth and metastasis through stromal modulation.

Pancreatic cancer treatment faces challenges due to drug resistance as well as liver metastasis. As a new strategy for treating pancreatic cancer, combination therapy is now available, but the dense mesenchymal barrier in the tumor tissue blocks drug delivery and impairs its therapeutic efficacy. To address this issue, we prepared an ATF peptide-decorated liposomal co-loaded with cisplatin and rapamycin (ATF@Pt/Rapa Lps), which targets both tumor cells and cancer-associated fibroblasts that express uPAR receptors. In tumor sphere penetration experiments, ATF peptide modified liposomes significantly enhanced deep penetration. More importantly, the ATF@Pt/Rapa Lps disrupted the stroma, as demonstrated by the downregulation of ɑ-SMA, I collagen, and fibronectin protein in vivo and in vitro. In this way, highly effective drug delivery to tumor cells can be achieved. As expected, there was a stronger inhibition of cell proliferation and migration by ATF@Pt/Rapa Lps in vitro compared to free Pt/Rapa and Pt/Rapa Lps. Furthermore, ATF@Pt/Rapa Lps showed greater therapeutic effects in PANC02 transplanted tumor mice and liver metastasis mice models. Ultimately, multi-targeting nanomedicines co-loaded with Rapa and cisplatin may provide a new approach to treating metastatic pancreatic cancer.

Extraction and decontamination of microplastics from high organic matter soils: A simple, cost-saving and high efficient method.

This article introduces a simple, cost-saving and high efficient for the extraction and separation of microplastics (MPs) from soil with a high organic matter (SOM) content. In this study, MP with particle sizes of 154-600 µm of polyethylene (PE), polypropylene (PP), polystyrene (PS), polyvinyl chloride (PVC) and polyethylene terephthalate (PET) were artificially added into the five Mollisols with the high SOM. Three flotation solutions were used to extract these MPs from soils, and four digestion solutions were used to digest the SOM. As well, their destruction effects on MPs were also examined. The results showed that the flotation recovery rates of PE, PP, PS, PVC and PET were 96.1%-99.0% by ZnCl2 solution, while were 102.0%-107.2% by rapeseed oil, were 100.0%-104.7% by soybean oil. The digestion rate of SOM was 89.3% by H2SO4:H2O2 (1:40, v:v) at 70 °C for 48 h, and this was higher than by H2O2 (30%), NaOH and Fenton's reagent. However, the digestion rate of PE, PP, PS, PVC and PET were 0.0%-0.54% by H2SO4:H2O2 (1:40, v:v), and this was lower than by H2O2 (30%), NaOH and Fenton's reagent. As well, the factors influencing on MP extraction was also discussed. Generally, the best flotation solution was ZnCl2 (ρ > 1.6 g cm-3) and the best digestion method was H2SO4:H2O2 (1:40, v:v) at 70 °C for 48 h. The optimal extraction and digestion method were verified by the known concentrations of MPs (recovery rate of MPs was 95.7-101.7%), and this method was also used to extract MPs from long-term mulching vegetable fields in Mollisols of Northeast China.

Isoorientin reverses lung cancer drug resistance by promoting ferroptosis via the SIRT6/Nrf2/GPX4 signaling pathway.

Cisplatin, or DDP, is a highly successful and well-known chemotherapy drug used to treat cancer. Acquired resistance to chemotherapy is a major clinical concern, yet the mechanisms of this resistance are still unknown. Ferroptosis is a type of cell death distinct from other forms, fueled by a buildup of iron-associated lipid reactive oxygen species (ROS). Gaining insight into the process of ferroptosis could lead to novel treatments for overcoming cancer resistance. In this study, the combination of isoorientin (IO) and DDP treatment resulted in a significant decrease in the viability of drug-resistant cells, a substantial increase in intracellular iron, malondialdehyde (MDA) and ROS concentrations, a notable decrease in glutathione concentration, and the occurrence of ferroptosis in cells, as revealed by in vitro and in vivo experiments. Additionally, there was a decrease in the expression of nuclear factor-erythroid factor 2-related factor 2 (Nrf2), glutathione peroxidase 4 (GPX4), and sirtuin 6 (SIRT6) proteins, and an increase in cellular ferroptosis. Isoorientin acts as a mediator to regulate cellular ferroptosis and reverse drug resistance in lung cancer cells by controlling the SIRT6/Nrf2/GPX4 signaling pathway. The findings of this study suggest that IO can promote ferroptosis and reverse drug resistance in lung cancer through the SIRT6/Nrf2/GPX4 signaling pathway, thus offering a theoretical basis for its potential clinical application.

Rapamycin Liposomes Combined with 5-Fluorouracil Inhibits Angiogenesis and Tumor Growth of APC (Min/+) Mice and AOM/DSS-Induced Colorectal Cancer Mice.

Background: Transgenic C57BL/6-APC(Min/+) spontaneous cancer mouse model and the Azoxymethane (AOM)/Dextran Sulfate Sodium (DSS) chemically induced orthotopic colorectal cancer mouse model represented distinct pathogenesis of colorectal cancers. Our previous study revealed that the combination of Rapamycin liposomes (Rapa/Lps) and 5-Fluorouracil (5-FU) has anti-colorectal cancer effects. However, the therapeutic efficacy of Rapa/Lps and 5-FU in other colorectal cancer mice models is yet to be thoroughly explored. The purpose of this study was to investigate the anti-tumor effect of Rapa/Lps combined with 5-FU in vivo and in vitro. Methods: In this study, we evaluated the effect of Rapa/Lps and 5-FU on APC (Min/+) mice and AOM/DSS-induced colorectal cancer mice. The small intestine, colorectum, serum, and plasma of mice in each group were collected following sacrifice to record the number of tumors. HE staining was utilized for observing pathological damage to intestine tissues. Tube formation assay, Transwell assay, wound healing assay, Western Blot were used to explore the anti-angiogenesis effect of drugs in HUVECs. Results: As expected, Rapa/Lps and 5-FU significantly suppressed tumor formation, decreased the number of tumors, and tumor load both in two mouse models, and had no influence on mouse weight. Mechanically, the anti-tumor effect of the drug also was associated in inhibiting angiogenesis and proliferation. Furthermore, we found that Rapa/Lps obviously inhibited HUVECs tube formation and migration. Conclusion: Altogether, we revealed the Rapa/Lps synergism with 5-FU decreased colon and small intestinal tumorigenesis in AOM/DSS-treated and APC (Min/+) mice, respectively, and correlated with anti-angiogenesis.

Heterogeneity of plastic residue was determined by both mulch film and external plastic pollutants in the farmland of Northeast China.

Plastic pollution in farmland ecosystems has been widely concerned. However, the heterogeneity and driving mechanisms of plastic residue (PR) remain unclear in the farmland surrounded by complex pollution points. In this study, the abundance, mass, and accumulation areas of PR of mulch film (MF) and non-MF (NMF) were investigated in a large area of the vegetable field covered by plastic mulching in a long-term in Northeast China. Geostatistics combined with classical statistics were used to clarify how pollution source and migration factors change the PR heterogeneity in the farmlands. Results indicated that the MF type was only polyethylene (PE) (79.1 % of total PR), while NMF accounted for 20.9 % of total PR. As well, NMF-polypropylene (PP) and NMF-PE accounted for 45.3 % and 39.7 % of total NMF respectively, followed by polystyrene accounting for 7.5 %. In the 0-20 cm soil layer, the spatial autocorrelation of mass and accumulation areas of MF were significantly (p < 0.05) positive, and their spatial pattern tended to cluster. The accumulation areas of MF was predoniment in northwest and southeast near the roadside in all soil layers, while the accumulation areas of NMF was higher near the landfill in the 0-20 cm soil layer. Landfill and residential areas were critical sources of PR for the farmland. Southwest wind and southeast wind were the main driving force of PR migration and their annual migration rates were 23.7 and 19.8 m·year-1. The functional groups on the surface of plastics were different after degradation (including different types and different utilization methods), and PR was oxidized could release or adsorb toxic substances from the soils. Generally, in order to reduce PR pollution, in addition to source control and recycling, farmland tillage should be avoided in the weather when the wind speed is strong, especially near the PR pollution source.

Field management changes the distribution of mesoplastic and macroplastic in Mollisols of Northeast China.

Mesoplastic (MaP) and macroplastic (MeP) coming from plastic mulching tend to cause negative effects on biota in ecosystems. However, it is still not clear how field management influences the distribution of MeP/MaP in soils. In this study, MeP/MaP was investigated in 0-20 and 20-30 cm soil layers of three vegetable fields (3.4-6.5 ha) after 13 years plastic-mulching in Mollisols of Northeast China under different management methods (MM) of fertilization and tillage frequency. The tillage frequency was MM2 > MM1 > MM3, while the fertilization was MM1 > MM2 > MM3. The results showed that polyethylene (PE), polypropylene (PP), polystyrene, polyvinyl chloride, polyethylene terephthalate (PET), polyamide, melamine-formaldehyde resin and polyether urethane were found in soil, and PE (>83.76%, from plastic mulching) was the predominant type of MeP/MaP. MeP abundance was significantly (p < 0.05) higher in MM1 and MM2 than that in MM3 in the 0-20 cm soil layer. MM1 and MM2 had the highest abundance of MeP/MaP of size <4 cm2 and 4-16 cm2, while MM3 had the highest abundance at the size >16 cm2. The broken index of MeP/MaP was significantly (p < 0.05) lower in MM2 compared with MM1 and MM3 in the 20-30 cm soil layer. Both tillage frequency and fertilization accelerate the breaking of plastics, especially since the influence was stronger from fertilization. Compared with original plastics, the PE, PP and PET's carbonyl index was significantly (p < 0.05) higher in the three MMs. Generally, fertilization and frequent tillage can reduce the physical effects of large-sized plastic debris on crop growth and increases the negative effects of small-sized plastic and new pollutants formed on biota in the agroecosystems. MeP/MaP recycling should be strengthened, and the irrigation and rotation of farmland should be carried out when the wind speed is weak to avoid plastic invasion.

Crops Change the Morphology, Abundance, and Mass of Microplastics in Mollisols of Northeast China.

Degradation of microplastics (MPs) by both physicochemical and biological processes in the natural environment is determined by the enzymes inside the soil, and which was severely influenced by crop growth and straw amendment (SA). However, it is still unclear how crop growth and SA influence degradation of MPs in soils. In this study, both catalase and sucrase were measured, and the stereomicroscope combined with microscopic infrared spectroscopy and scanning electron microscope (SEM) was used to detect the morphology and quantity of low-density polyethylene microplastic (LDPE-MP) and low-density polypropylene microplastic (LDPP-MP), after crop growth (maize and soybean, with and without SA, 1 and 2% MP) in an outdoor pot experiment, in the Mollisols. The results showed that the growth of the crops changed the morphology, functional groups (e.g., methylene, carbonyl), total mass, and abundance ratio of MPs of different sizes. These were possibly caused by enzymes that were significantly influenced by crop types, abundance, and types of MPs in the soils. Maize growth decreased the mass of LDPE-MP and LDPP-MP by 28.7 and 32.7%, respectively, and 2% (w/w) of LDPP-MP addition in soil decreased mass of 9%, which was higher than that in 1% (w/w) LDPP-MP addition in soil. Soybean growth with SA decreased the mass of LDPE-MP and LDPP-MP by 36.6 and 20.7%, respectively, than the control treatment (CK). Compared with CK, both crop growth and SA changed the abundance of MPs of different sizes and decreased the mean size of MPs. The LDPE-MP could be more easily degraded by enzymes in the soils compared to LDPP-MP when the MP size was smaller with surface roughness. Generally, both maize and soybean growth can accelerate MP change in soils, and MP change process was mainly determined by SA, MP types, and the dose effect of MP.

Molecularly Imprinted Polymers with Enzymatic Properties Reduce Cytokine Release Syndrome.

A core-shell molecularly imprinted polymer nanoparticle with biological enzyme functional characteristics was developed by oxidative polymerization of template protein and polydopamine on the surface of protease-copper phosphate hybrid nanoflowers by molecular imprinting technology and enzyme immobilization technology. The obtained molecularly imprinted polymer showed specific binding characteristics with the template protein. It recognized and enriched the target molecules through the surface molecularly imprinted sites of the shell structure. In addition, the bound target molecules were further degraded into fragments by nanozymes with biological enzyme characteristics in the core. In this study, molecular imprinting technology and biotechnology were combined to obtain bifunctional molecularly imprinted polymer nanoparticles that can not only enrich template molecules but also degrade them into fragments. Herein, we selected interleukin 6 (IL-6), the target molecule of cytokine release syndrome (CRS), as a template molecule, and reported a molecularly imprinted polymer with degrading enzyme properties that can rapidly reduce IL-6 levels in vivo, including a molecularly imprinted layer that can recognize and bind IL-6 and nanozymes that can degrade IL-6 and deactivate it. It is used to clear the excessive secretion of IL-6 in CRS and reduce the level of IL-6 in the body to achieve the purpose of adjuvant treatment of CRS.

Myrothecol A, a new alkylresorcinol with cytotoxicity from Myrothecium sp.

A new alkylresorcinol, myrothecol A (1), along with two known ones (2 and 3), were isolated from a fungal strain Myrothecium sp. GY170016. Their structures were elucidated by extensive spectroscopic analysis. The absolute configuration of 1 was determined by electronic circular dichroism experiment. This is the first case of the presence of alkylresorcinols in genus Myrothecium. All the isolates were evaluated for their cytotoxic activities against human cancer cell line MCF-7 with IC50 values of 16.7, 13.2, 21.3 µM, respectively.

Delivery of Rapamycin by Liposomes Synergistically Enhances the Chemotherapy Effect of 5-Fluorouracil on Colorectal Cancer.

BACKGROUND: Rapamycin is a promising agent for treating tumors, but clinical applications of rapamycin are limited due to its poor water solubility and low bioavailability. This paper constructs a liposome delivery system for rapamycin to improve the effect in treating colorectal cancer. METHODS: We prepared the rapamycin liposomes using the ethanol injection method. The cellular uptake and biodistribution were detected by LC-MS and in vivo imaging system. MTT assay, transwell migration experiment, flow cytometry, and Western blot analysis evaluated the antitumor effect of rapamycin liposomes in vitro. Furthermore, HCT-116 tumor-bearing mice were used to assess the therapeutic efficacy of rapamycin liposomes in vivo. RESULTS: The prepared rapamycin liposomes had a particle size of 100±5.5 nm and with a narrow size distribution. In vitro cellular uptake experiments showed that the uptake of rapamycin liposomes by colorectal cells was higher than that of free rapamycin. Subsequently, in vivo imaging experiments also demonstrated that rapamycin liposomes exhibited higher tumor accumulation. Therefore, the ability of rapamycin liposomes to inhibit tumor proliferation, migration and to induce tumor apoptosis is superior to that of free rapamycin. We also demonstrated in vivo good antitumor efficacy of the rapamycin liposomes in HCT-116 xenograft mice. In addition, rapamycin liposomes and 5-FU can synergistically improve the efficacy of colorectal cancer via the Akt/mTOR and P53 pathways. CONCLUSION: Collectively, rapamycin liposomes are a potential treatment for colorectal cancer, as it not only improves rapamycin's antitumor effect but also synergistically enhances 5-FU's chemotherapy effect.

Non-biodegradable microplastics in soils: A brief review and challenge.

Non-biodegradable microplastics (MPs) pollution long-termly existed in soils, and was only concerned in recent years. In order to better understand MP behavior in soils, the sources, migration, distribution, biological effects, degradation and analytical methodology of non-biodegradable MPs in soils were quantificationally summarized from 170 publications based on Web of Science in 1950-2020. From the publications, we found these studies were mainly carried out in the Asia (60.0%) and Europe (23.3%), and most were on agricultural soils (68.5%). Polyethylene-MP (78.8% of the studies), Polypropylene-MP (78.8%), and Polystyrene-MP (45.5%) were the MPs most frequently found in the soils, with a MP size of 20-5000 µm being most common. Of the soil samples 64.3% contained MP 1000-4000 items kg-1, and the colour frequency ranking is blue (66.7%) > white (61.1%) ≈ red ≈ black. MPs changed the soil microenvironment and microorganism activity, and caused the negative effects on both soil animals (100%) and plants (57.9%). MP degradation was influenced by the photooxidation reactions, microorganism activities, enzymatic effects, environmental conditions, and by the composition, size and morphology of the MPs. An optional analytical method was suggested in this study. At the end of paper, the urgent and important research work in the future was prospected.

T-cell death-associated gene 8 accelerates atherosclerosis by promoting vascular smooth muscle cell proliferation and migration.

BACKGROUND AND AIMS: Atherosclerosis is a serious cardiovascular disease, featuring inflammation, abnormal proliferation and migration of vascular smooth muscle cells (VSMCs). During atherosclerosis, inflammation may cause low pH. T-cell death-associated gene 8 (Tdag8) is a proton-sensing receptor, however, the role of Tdag8 in VSMCs remains unknown. This study aimed to investigate the potential effects of Tdag8 in VSMCs during atherosclerosis. METHODS: We examined the expression of Tdag8 in an atherosclerotic model of high-fat-diet-fed ApoE-/- mice, while the role and mechanism of Tdag8 in phenotype transformation, proliferation and migration of VSMCs were investigated in a series of in vivo and in vitro experiments. RESULTS: We first found that Tdag8 expression at the mRNA and protein level was significantly increased in atherosclerotic ApoE-/- mice. Immunofluorescence staining showed that Tdag8 was primarily distributed in PCNA-positive VSMCs and the phenotype of VSMCs switching from contractile phenotype to synthetic phenotype. Additionally, the protein level of Tdag8 was upregulated in FBS-treated VSMCs. VSMCs proliferation and migration were inhibited by Tdag8 silencing and increased by Tdag8 overexpression. Further mechanistic studies showed that cAMP level was increased in Tdag8-overexpressing VSMCs and ApoE-/- mice. However, the PKA inhibitor H-89 reversed Tdag8-induced VSMC proliferation and migration. CONCLUSIONS: The results demonstrate that Tdag8 mediated phenotype transformation, proliferation and migration of VSMCs via the cAMP/PKA signaling pathway, thus partially contributing to atherosclerosis.

Nox1/4 dual inhibitor GKT137831 attenuates hypertensive cardiac remodelling associating with the inhibition of ADAM17-dependent proinflammatory cytokines-induced signalling pathways in the rats with abdominal artery constriction.

NADPH oxidases (Noxs) 1/4 dual inhibitor GKT137831 prevents hypertensive cardiac remodelling in angiotensin II-infused transgenic mice with cardiomyocyte-specific human Nox4 (c-hNo x 4 Tg); however, further research is still required to determine the beneficial role of GKT137831 in hypertensive cardiac remodelling in other types of hypertensive models because this hypertensive model is insufficient to mimic the complicated pathological mechanisms of hypertension. A disintegrin and metalloprotease 17 (ADAM17) promotes the shedding of tumour necrosis factor α (TNF-α), TNF-α receptor, interleukin 1 receptor-II and interleukin 6 (IL-6) receptor from cells, thereby mediating the signalling pathways induced by corresponding proinflammatory cytokines. This study aimed to determine whether GKT137831 prevents hypertensive cardiac remodelling and its mechanisms of action in the rats with abdominal artery coarctation (AAC). The rats subjected to AAC were orally given GKT137831 for a consecutive period of 28 days. Echocardiography and histological analysis were performed to evaluate cardiac remodelling; and immunohistochemistry and real-time PCR were used to detect the expression of proinflammatory cytokines. GKT137831 significantly suppressed hypertensive cardiac remodelling in AAC-induced hypertensive rats. Concurrently, Nox1/4 dual inhibitor GKT137831 reduced the protein and mRNA levels of proinflammatory cytokines interleukin 1ß (IL-1ß), IL-6, and TNF-α in the left ventricle of AAC-induced hypertensive rats. Moreover, the treatment with GKT137831 markedly diminished the protein and mRNA levels of ADAM17 in the left ventricle of AAC-induced hypertensive rats. In summary, Nox1/4 dual inhibitor GKT137831 protects against hypertensive cardiac remodelling in AAC-induced hypertensive rats, and the inhibition of ADAM17-dependent proinflammatory cytokines-induced signalling pathways are related to its beneficial effect on hypertensive cardiac remodelling.

Knockdown of zinc transporter ZIP8 expression inhibits neuroblastoma progression and metastasis in vitro.

Neuroblastoma is one of the leading causes of cancer­associated mortality worldwide, particularly in children, partially due to the absence of effective therapeutic targets and diagnostic biomarkers. Therefore, novel molecular targets are critical to the development of therapeutic approaches for neuroblastoma. In the present study, the functions of zinc transporter ZIP8 (Zip8), a member of the zinc transporting protein family, were investigated as novel molecular targets in neuroblastoma cancer cells. The proliferation rates of neuroblastoma cancer cells were significantly decreased when Zip8 was knocked down by lentiviral­mediated RNA interference. Study of the molecular mechanism suggested that Zip8 modulated the expression of key genes involved in the nuclear factor­κB signaling pathway. Furthermore, Zip8 depletion suppressed the migratory potential of neuroblastoma cancer cells by reducing the expression levels of matrix metalloproteinases. In conclusion, the results of the present study suggested that Zip8 was an important regulator of neuroblastoma cell proliferation and migration, indicating that Zip8 may be a potential anticancer therapeutic target and a promising diagnostic biomarker for human neuroblastoma.

Polymorphisms and haplotypes of the TGF-ß1 gene are associated with risk of polycystic ovary syndrome in Chinese Han women.

OBJECTIVES: Transforming growth factor-beta 1 (TGF-ß1) is one of the multifunctional cytokine families. It takes part in a series of physiological and pathological processes in the human body, including wound healing, tissue fibrosis and embryonic development. Recent studies have shown that TGF-ß1 participates in the development of polycystic ovary syndrome (PCOS). This study was therefore designed to investigate the association of TGF-ß1 polymorphism with the risk of PCOS. STUDY DESIGN: We enrolled 328 PCOS patients and 358 healthy individuals in this study. Five single nucleotide polymorphisms (SNPs) - rs4803457C/T, rs11466313 deletion/AGG, rs2217130C/T, rs1800469C/T and rs1800470C/T - were detected using Snapshot technology. Linkage disequilibrium and haplotype analysis was conducted among the five SNPs. The relationship between genotypes and haplotypes and the risk of PCOS was also explored. RESULTS: The TT/CT/CC genotype frequencies of rs4803457 in the PCOS group and the control group were 0.2805/0.4878/0.2317 and 0.3659/0.4749/0.1592 respectively. The C/T allele frequencies in the PCOS group and control group were 0.3813/0.6187 and 0.3966/0.6034 respectively. There were significant differences in genotype distribution frequencies and allele frequencies between these two groups (P=0.018). Logistic regression analysis showed that CC genotype had higher risk of PCOS than the no CC genotype in rs4803457 loci (OR=1.75, 95%CI=1.11-2.75). Haplotype analysis further showed that the haplotypes "T-del-C-C-C", "C-del-C-C-C" and "C-del-C-T-C" were associated with the highest risk of PCOS. However, for rs11466313 deletion/AGG, rs2217130C/T, rs1800469C/T and rs1800470C/T, no significant association with PCOS risk was observed. CONCLUSION: The TGF-ß1 gene rs4803457C/T polymorphism is associated with susceptibility to PCOS, and is the key contributor for the development of PCOS in Chinese Han women. The haplotypes T-del-C-C-C, C-del-C-C-C and C-del-C-T-C are also risk factors for PCOS susceptibility among Chinese Han women.

Meta-analysis of estradiol for luteal phase support in in vitro fertilization/intracytoplasmic sperm injection.

OBJECTIVE: To evaluate whether the addition of E(2) for luteal phase support (LPS) in IVF/intracytoplasmic sperm injection (ICSI) could improve the outcome of clinical pregnancy. DESIGN: Meta-analysis. SETTING: University hospital center. PATIENT(S): Women underwent IVF or ICSI using the GnRH agonist or GnRH antagonist protocol. INTERVENTION(S): Progesterone alone or combined with E(2) for LPS. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate per patient (CPR/PA), clinical pregnancy rate per ET, implantation rate, ongoing pregnancy rate per patient, clinical abortion rate, and ectopic pregnancy rate. RESULT(S): Fifteen relevant randomized controlled trials (RCTs) were identified that included a total of 2,406 patients. There was no statistical difference between E(2) + P group and P-only group regarding the primary outcome of CPR/PA for different routes of administration of E(2) (oral, vaginal, and transdermal) or other relevant outcome measures. No significant effect was observed for different daily doses of E(2) (6, 4, and 2 mg), even through oral medication in CPR/PA. CONCLUSION(S): The best available evidence suggests that E(2) addition during the luteal phase does not improve IVF/ICSI outcomes through oral medication, even with different daily doses. Furthermore, RCTs that study other administration routes are needed.

HIV-infection resistance in PMBC-derived dendritic cells modified with recombinant virus.

This study aimed to identify the characteristics of recombinant-adenovirus-modified PBMC-derived dendritic cells and their resistance to HIV-1 infection by integrating the CCR5∆32, CCR5siRNA, HIV-1 pol and HIV-1 int genes into a recombinant adenovirus vector using the AdEasy system. Dendritic cells (DCs) were isolated from human PBMCs from blood of healthy donors. The expression of CCR5∆32, CCR5, CXCR4 and HIV-1 p24 in PBMCs or modified cells was measured by western blot, p24 expression in cell lysates was measured by ELISA, and HIV-1 entry was measured by ß-galactosidase assay. Furthermore, T-cell immunity induced by the recombinant adenovirus was measured by ELISPOT assay. After the cells were modified by Ad-R5∆32siRNA, the expression of CCR5∆32 increased, while the expression of CCR5 and CXCR4 decreased. There was no adverse effect of adenoviral gene transfer on DC development. CD83 expression on the surface of mature DCs did not change after gene transfer. The expression of p24 remained at low levels in modified cells when challenged by HIV-1. The modified cells showed resistance to HIV-1 infection. Results indicated that recombinant-adenovirus-modified cells demonstrated good resistance to HIV-1 infection. Modification of HSC-derived immune cells, such as DCs, may be a potent strategy to resist HIV-1 infection.