Disruption of Super-Enhancers in Activated Pancreatic Stellate Cells Facilitates Chemotherapy and Immunotherapy in Pancreatic Cancer.

One major obstacle in the drug treatment of pancreatic ductal adenocarcinoma (PDAC) is its highly fibrotic tumor microenvironment, which is replete with activated pancreatic stellate cells (a-PSCs). These a-PSCs generate abundant extracellular matrix and secrete various cytokines to form biophysical and biochemical barriers, impeding drug access to tumor tissues. Therefore, it is imperative to develop a strategy for reversing PSC activation and thereby removing the barriers to facilitate PDAC drug treatment. Herein, by integrating chromatin immunoprecipitation (ChIP)-seq, Assays for Transposase-Accessible Chromatin (ATAC)-seq, and RNA-seq techniques, this work reveals that super-enhancers (SEs) promote the expression of various genes involved in PSC activation. Disruption of SE-associated transcription with JQ1 reverses the activated phenotype of a-PSCs and decreases stromal fibrosis in both orthotopic and patient-derived xenograft (PDX) models. More importantly, disruption of SEs by JQ1 treatments promotes vascularization, facilitates drug delivery, and alters the immune landscape in PDAC, thereby improving the efficacies of both chemotherapy (with gemcitabine) and immunotherapy (with IL-12). In summary, this study not only elucidates the contribution of SEs of a-PSCs in shaping the PDAC tumor microenvironment but also highlights that targeting SEs in a-PSCs may become a gate-opening strategy that benefits PDAC drug therapy by removing stromal barriers.

Self-assembly of CXCR4 antagonist peptide-docetaxel conjugates for breast tumor multi-organ metastasis inhibition.

As a representative chemotherapeutic drug, docetaxel (DTX) has been used for breast cancer treatment for decades. However, the poor solubility of DTX limits its efficacy, and the DTX based therapy increases the metastasis risk due to the upregulation of C-X-C chemokine receptor type 4 (CXCR4) expression during the treatment. Herein, we conjugated CXCR4 antagonist peptide (CTCE) with DTX (termed CTCE-DTX) as an anti-metastasis agent to treat breast cancer. CTCE-DTX could self-assemble to nanoparticles, targeting CXCR4-upregulated metastatic tumor cells and enhancing the DTX efficacy. Thus, the CTCE-DTX NPs achieved promising efficacy on inhibiting both bone-specific metastasis and lung metastasis of triple-negative breast cancer. Our work provided a rational strategy on designing peptide-drug conjugates with synergistic anti-tumor efficacy.

The Combination of Sinusoidal Perfusion Enhancement and Apoptosis Inhibition by Riociguat Plus a Galactose-PEGylated Bilirubin Multiplexing Nanomedicine Ameliorates Liver Fibrosis Progression.

Chronic liver injury and continuous wound healing lead to extracellular matrix (ECM) deposition and liver fibrosis. The elevated production of reactive oxygen species (ROS) in the liver leads to the apoptosis of hepatocytes and the activation of hepatic stellate cells (HSCs). In the current study, we describe a combination strategy of sinusoidal perfusion enhancement and apoptosis inhibition enabled by riociguat together with a tailor-designed galactose-PEGylated bilirubin nanomedicine (Sel@GBRNPs). Riociguat enhanced sinusoidal perfusion and decreased the associated ROS accumulation and inflammatory state of the fibrotic liver. Concurrently, hepatocyte-targeting galactose-PEGylated bilirubin scavenged excessive ROS and released encapsulated selonsertib. The released selonsertib inhibited apoptosis signal-regulating kinase 1 (ASK1) phosphorylation to alleviate apoptosis in hepatocytes. The combined effects on ROS and hepatocyte apoptosis attenuated the stimulation of HSC activation and ECM deposition in a mouse model of liver fibrosis. This work provides a novel strategy for liver fibrosis treatment based on sinusoidal perfusion enhancement and apoptosis inhibition.

Restoration of Sinusoid Fenestrae Followed by Targeted Nanoassembly Delivery of an Anti-Fibrotic Agent Improves Treatment Efficacy in Liver Fibrosis.

During the onset of liver fibrosis, capillarized liver sinusoidal endothelial cells (LSECs) limit substance exchange between the blood and the Disse space, further accelerating hepatic stellate cell (HSCs) activation and fibrosis progression. Limited accessibility of therapeutics to the Disse space is often overlooked and remains a major bottleneck for HSCs-targeted therapy in liver fibrosis. Here, an integrated systemic strategy for liver fibrosis treatment is reported, utilizing pretreatment with the soluble guanylate cyclase stimulator, riociguat, followed by insulin growth factor 2 receptor-mediated targeted delivery of the anti-fibrosis agent, JQ1, via peptide-nanoparticles (IGNP-JQ1). The riociguat reversed the liver sinusoid capillarization to maintain a relatively normal LSECs porosity, thus facilitating the transport of IGNP-JQ1 through the liver sinusoid endothelium wall and enhancing the accumulation of IGNP-JQ1 in the Disse space. IGNP-JQ1 is then selectively taken up by activated HSCs, inhibiting their proliferation and decreasing collagen deposition in the liver. The combined strategy results in significant fibrosis resolution in carbon tetrachloride-induced fibrotic mice as well as methionine-choline-deficient-diet-induced nonalcoholic steatohepatitis (NASH) mice. The work highlights the key role of LSECs in therapeutics transport through the liver sinusoid. The strategy of restoring LSECs fenestrae by riociguat represents a promising approach for liver fibrosis treatment.

Biomimetic Nanoparticles Carrying a Repolarization Agent of Tumor-Associated Macrophages for Remodeling of the Inflammatory Microenvironment Following Photothermal Therapy.

The complete regression of residual tumors after photothermal therapy (PTT) depends on the activation and recognition of the immune system. However, the inevitable local inflammation after PTT in residual tumor recruits abundant abnormal immune cells, especially the tumor-associated macrophages (TAMs) which further promote immune escape and survival of the remaining tumor cells, resulting in the tumor recurrence and progression. To solve this problem, herein we explored biomimetic nanoparticles carrying repolarization agent of TAMs to remodel the post-PTT inflammatory microenvironment. The polydopamine nanoparticles were used simultaneously as photothermal transduction agents to ablate tumor cells and the delivery vehicles for TMP195 which can repolarize the M2-like TAMs into an antitumor phenotype. In addition, a biomimetic decoration of macrophage membrane coating was designed to endow nanoparticles the ability to actively target the tumor site after PTT mediated by inflammation-mediated chemotaxis. In the breast tumor model, these biomimetic nanoparticles with immune-modulating ability significantly elevated the levels of M1-like TAMs, ultimately resulting in a tumor-elimination rate of 60%, increased from 10% after PTT. This synergistic treatment strategy of PTT and TAMs repolarization provides a promising approach to address the deteriorated tumor microenvironment after PTT and proposes a more effective way for combinational treatment option in clinic.

Effects of early second-look hysteroscopy combined with intrauterine balloon dilatation on reproductive outcomes for women with intrauterine adhesions.

OBJECTIVE: To investigate the effect of early second-look office hysteroscopy combined with intrauterine balloon dilatation on prognosis and pregnancy rate for women with intrauterine adhesions. METHODS: A retrospective analysis of 156 women diagnosed with intrauterine adhesions by hysteroscopy at Shenyang Women's and Children's Hospital, China, from April 2017 to January 2019. The study women underwent intrauterine balloon dilatation 10 days after transcervical resection of adhesion (TCRA) and hysteroscopy 20 days after TCRA (n=81). The control women underwent hysteroscopy 3 months after TCRA (n=75). Estrogen and aspirin were routinely administered postoperatively to all women. Data, including American Fertility Society (AFS) scores assessed by hysteroscopy, endometrial thickness measured by ultrasound, and menstruation and pregnancy outcomes assessed by interview, were compared between the two groups. RESULTS: The degree of intrauterine adhesions, menstrual status, and endometrial thickness were improved in both groups after TCRA. Greater improvement in AFS score, menstruation, and endometrial thickness was observed in the study group than in the control group. After follow-up, more women in the study group achieved pregnancy (48.1% vs 30.7%, P<0.05). CONCLUSION: Early second-look of hysteroscopy combined with intrauterine balloon dilatation after hysteroscopic TRCA might improve the prognosis and postoperative pregnancy rate for women with intrauterine adhesions.

[Identification of cryptic structural chromosomal aberrations in parents through detection of copy number variations in miscarriage tissues].

OBJECTIVE: To explore the genetic cause for abnormal pregnancies through detecting chromosomal copy number variations (CNVs) in abortic tissues by next generation sequencing (NGS). METHODS: NGS technique was used to detect CNVs in abortion tissues. Parental chromosomal karyotypes were predicted based on the results. The aberrant chromosomal segments of the parents were accurately mapped by G-banding karyotyping analysis and fluorescence in situ hybridization (FISH). RESULTS: In addition to numerical chromosomal aberrations, 12 microdeletion/microduplications were detected by NGS. For 8 families where both parents accepted chromosomal karyotyping, 4 carriers of chromosomal abnormalities were identified. One marker chromosome was missed by karyotyping analysis, and a mother was confirmed to carry a cryptic balanced translocation by FISH. CONCLUSION: NGS can facilitate detection of cryptic chromosomal translocations in couples with repeated pregnancy failure and is of great value for detecting abnormal CNVs for its high sensitivity.

PAHs pass through the cell wall and partition into organelles of arbuscular mycorrhizal roots of ryegrass.

The subcellular process and distribution of polycyclic aromatic hydrocarbons (PAHs) in arbuscular mycorrhizal plants remains to be elucidated. This work used a greenhouse experiment to show that, accompanied by the apoplastic and symplastic water movement through the root, acenaphthene (ACE) as a representative PAH passed through the cell-wall boundary, dissolved in the cell solution, and partition organelles in arbuscular mycorrhizal roots of ryegrass (Lolium multiflorum Lam.). The observed concentrations of ACE in organelles were 0.6 to 4.4 times higher than in the cell walls. The cell wall and organelles were the dominant storage domains for ACE in the root, and the distribution of ACE in cells of mycorrhizal ryegrass roots was, in descending order, cell organelles (40.8-70.8%) > cell wall (19.7-3.8%) cell solution (9.6-20.5%).

Arbuscular mycorrhizal fungal hyphae contribute to the uptake of polycyclic aromatic hydrocarbons by plant roots.

The arbuscular mycorrhizal (AM) hyphae-mediated uptake of polycyclic aromatic hydrocarbons (PAHs) by the roots of ryegrass (Lolium multiflorum Lam.) was investigated using three-compartment systems. Glomus mosseae and Glomus etunicatum were chosen, and fluorene and phenanthrene were used as representative PAHs. When roots were grown in un-spiked soils, AM hyphae extended into PAH-spiked soil and clearly absorbed and transported PAHs to roots, resulting in high concentrations of fluorene and phenanthrene in roots. This was further confirmed by the batch equilibration experiment, which revealed that the partition coefficients (K(d)) of tested PAHs by mycorrhizal hyphae were 270-356% greater than those by roots, suggesting the great potential of hyphae to absorb PAHs. Because of fluorene's lower molecular weight and higher water solubility, its translocation by hyphae was greater than that of phenanthrene. These results provide new perspectives on the AM hyphae-mediated uptake by plants of organic contaminants from soil.

[Level of urocortin mRNA during labor and effect of urocortin on myometrial contractility in vitro].

OBJECTIVE: To examine the expression of urocortin mRNA during labor and the effect of urocortin on myometrial contractility, and to investigate its role in the onset and progress of labor. METHODS: (1) Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), using beta-actin as internal standard was applied to determine the levels of urocortin mRNA in human placenta and myometrium from the group of cesarean section before (10 cases) and during (10 cases in latent phase and 10 cases in active phase) labor. (2) The isolated myometrial strips of pregnant women (n = 24) were prepared. The effects of urocortin with or without prostaglandin F(2alpha) (PGF(2alpha)) and oxytocin on myometrial contractility were evaluated by areas under the curve. RESULTS: (1) Semi-quantitative RT-PCR showed that the expression level of urocortin mRNA in placenta and myometrium after the onset of labor were higher than before labor (1.23 +/- 0.52, 1.32 +/- 0.22; 0.83 +/- 0.38, 0.94 +/- 0.13, respectively, P < 0.05). (2) Urocortin itself did not affect myometrial tension development at all concentrations tested, but it markedly increased PGF(2alpha)-induced myometrial contractility. The average area under the curve of controls was (2.12 +/- 0.15) cm(2) and of study strips was (3.90 +/- 0.33) cm(2) (P < 0.05); urocortin did not increase the myometrial response to oxytocin compared with controls (P > 0.05). CONCLUSION: The study indicates urocortin may indirectly modulate myometrial contractility during labor.

[Study on the level of placental urocortin and corticotropin-releasing hormone receptor 2beta of preeclampsia].

OBJECTIVE: To determine the relation between the expression of urocortin and corticotropin-releasing hormone receptor 2beta (CRH-R2beta) in the placenta and pathogenesis of preeclampsia. METHODS: Placentas were collected from 20 pregnant women with preeclampsia as study group and 20 normal pregnant women as control group. Urocortin mRNA and CRH-R2beta mRNA were measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Urocortin peptide was measured by immunohistochemistry. RESULTS: (1) The mRNA expression of urocortin was significantly higher (P < 0.05) in preeclampsia (1.14 +/- 0.26) compared with normal pregnancies (0.78 +/- 0.46). (2) There was no significant difference between the expression of CRH-R2beta mRNA of preeclampsia (0.89 +/- 0.33) and of normal pregnancies (0.93 +/- 0.50). (3) The result of immunohistochemistry demonstrated that urocortin was predominantly localized in syntrophoblast with weak staining in cyntrophoblast and capillaries. Urocortin in syntrophoblast of preeclampsia was significantly higher than that of normal pregnancy (P < 0.05). CONCLUSIONS: Our study demonstrates that urocortin peptide and mRNA is increased in women with preeclampsia. This may be a kind of stress-responsive compensatory mechanism in human placenta. Urocortin may play a role in the pathology of the disease.