Preferential Recruitment of Th17 Cells to Cervical Cancer via CCR6-CCL20 Pathway

<div><p>Our previous studies suggest that Th17 cells accumulate within tumor tissues and correlate with recurrence of cervical cancer patients. However, the source of the increased tumor-infiltrating Th17 cells remains poorly understood. We investigated the prevalence, phenotype and trafficking property of Th17 cells in patients with cervical cancer. Our results showed that Th17 cells highly aggregated within tumor tissues in an activated phenotype with markedly increased expression of CCR6. Correspondingly, level of CCL20 in the tumor tissues was significantly higher than that in non-tumor and normal control tissues, and strongly positively associated with Th17 cells. Further, in vitro migration assay showed CCL20 had effective chemotaxis to circulating Th17 cells. In conclusion, Th17 cells are recruited into tumor tissues preferentially through CCR6-CCL20 pathway, which can serve as a novel therapeutic target for cervical cancer.</p></div

Direct Imaging of Transmembrane Dynamics of Single Nanoparticles with Darkfield Microscopy: Improved Orientation Tracking at Cell Sidewall

Investigation of the cellular internalization processes of individual nanoparticles (NPs) is of great scientific interest with implications to drug delivery and NP biosafety. Herein, by using dual-channel polarization darkfield microcopy (DFM) and single gold nanorods (AuNRs) as orientation probes, we developed a method that is capable of monitoring AuNR orientation dynamics during its transmembrane process. With annular oblique illumination and a birefringent prism to split AuNR plasmonic scattering into two channels of orthogonal polarizations, the AuNR azimuth and polar angles are obtained from their intensity difference and intensity sum. By placing the focal plane of the microscope objective at the elevated cell sidewall rather than at the flat cell top, interference from cellular background is reduced and the signal-to-noise ratio of AuNR orientation sensing is improved significantly, especially for AuNRs inserting into the membrane at a large out-of-plane angle. As a result, we were able to capture the complete membrane-crossing dynamics of single AuNRs. This powerful method could be utilized to obtain valuable insights on NP endocytosis mechanisms of various cells

Chemokine effects on Th17 cell recruitment.

<p>Migration assays were performed in a Transwell system. (A) Th17 cells migrate in response to recombinant human CCL17, CCL20 and CCL22 in dose-dependent manner (n = 5). Specific antibodies to chemokines significantly inhibit Th17 cell migration.***<i>P</i> < 0.001. (B and C) Expression of CCL20 by HeLa, Siha, and C-33A cells detected using Real-time PRC (B) and ELISA (C). All the three cervical cells could highly secrete CCL20 with highest level of CCL20 by HeLa cells. (D) Th17 cells also migrate toward culture supernatants of HeLa, Siha, and C-33A cells, which can be efficiently blocked by antibody against CCL20 alone or in combination with CCL17 and CCL22, but significantly less effectively by antibody against CCL17 or CCL22 (n = 3). *<i>P</i> < 0.05, **<i>P</i> < 0.01,***<i>P</i> < 0.001.</p

Association of intratumoral Th17-cell prevalence with clinical parameters.

<p>Association of intratumoral Th17-cell prevalence with clinical parameters.</p

Phenotypic analysis of Th17 cells in patients with cervical cancer.

<p>(A) Representative expression profiles of CD45RO, HLA-DR, Granzyme B and PD-1 in tumor-infiltrating Th17 cells. The percentages represent the frequencies of various markers in Th17 cells. (B) Representative expression profiles of CCR4, CCR6 and CD49d on Th17 cells from peripheral blood (long dotted line), non-tumor (dotted line) and tumor tissues (solid line). The percentages represent the frequencies of various markers on tumor-infiltrating Th17 cells. (C) Statistical analysis of surface expression of CCR4, CCR6, CD49d on Th17 cells from peripheral blood, non-tumor and tumor tissues (n = 25). *<i>P</i> < 0.05, **<i>P</i> < 0.01, ***<i>P</i> < 0.001.</p

Th17 cells are highly enriched in tumors of patients with cervical cancer.

<p>Th17 cells were gated from CD3⁺ T cells by flow cytometry. (A) Representative IL-17 expression profiles in CD4⁺ T cells from the four studied groups. The percentages represent the frequency of Th17 cells among CD4⁺ T cells. (B) Statistical analysis show that the frequency of Th17 cells was higher in patients with cervical cancer, especially among tumor-infiltrating lymphocytes (n = 35). **<i>P</i> < 0.01, ***<i>P</i> < 0.001. (C) Representative images for Th17 cells (IL-17⁺) and Tregs (FoxP3⁺) infiltration in cervical cancer tissue from the same patient. Immunostained cells (brown, indicated by black arrow) and tumor cells (blue). Magnification,×100.</p

Table1_Advances in the research of sulfur dioxide and pulmonary hypertension.docx

Pulmonary hypertension (PH) is a fatal disease caused by progressive pulmonary vascular remodeling (PVR). Currently, the mechanisms underlying the occurrence and progression of PVR remain unclear, and effective therapeutic approaches to reverse PVR and PH are lacking. Since the beginning of the 21st century, the endogenous sulfur dioxide (SO2)/aspartate transaminase system has emerged as a novel research focus in the fields of PH and PVR. As a gaseous signaling molecule, SO2 metabolism is tightly regulated in the pulmonary vasculature and is associated with the development of PH as it is involved in the regulation of pathological and physiological activities, such as pulmonary vascular cellular inflammation, proliferation and collagen metabolism, to exert a protective effect against PH. In this review, we present an overview of the studies conducted to date that have provided a theoretical basis for the development of SO2-related drug to inhibit or reverse PVR and effectively treat PH-related diseases.</p

Synthesis of Pyrazolo[5,1‑<i>a</i>]isoindoles and Pyrazolo[5,1‑<i>a</i>]isoindole-3-carboxamides through One-Pot Cascade Reactions of 1‑(2-Bromophenyl)buta-2,3-dien-1-ones with Isocyanide and Hydrazine or Acetohydrazide

A novel and efficient method for the construction of the pyrazolo­[5,1-<i>a</i>]­isoindole scaffold via a one-pot three-component cascade reaction of 1-(2-bromophenyl)­buta-2,3-dien-1-one with hydrazine and isocyanide promoted by a Pd catalyst is described. This cascade process proceeds through initial condensation of the allenic ketone with hydrazine followed by Pd-catalyzed isocyanide insertion into the C–Br bond and intramolecular C–N bond formation. Interestingly, when acetohydrazide was used in place of hydrazine, a more sophisticated procedure involving condensation, isocyanide insertion into C–H and C–Br bonds, deacetylation, and formation of C–C, C–O, and C–N bonds occurred to afford pyrazolo­[5,1-<i>a</i>]­isoindole-3-carboxamides with good efficiency

Siderophore Biosynthesis Governs the Virulence of Uropathogenic Escherichia coli by Coordinately Modulating the Differential Metabolism

Urinary tract infections impose substantial health burdens on women worldwide. Urinary tract infections often incur a high risk of recurrence and antibiotic resistance, and uropathogenic E. coli accounts for approximately 80% of clinically acquired cases. The diagnosis of, treatment of, and drug development for urinary tract infections remain substantial challenges due to the complex pathogenesis of this condition. The clinically isolated UPEC 83972 strain was found to produce four siderophores: yersiniabactin, aerobactin, salmochelin, and enterobactin. The biosyntheses of some of these siderophores implies that the virulence of UPEC is mediated via the targeting of primary metabolism. However, the differential modulatory roles of siderophore biosyntheses on the differential metabolomes of UPEC and non-UPEC strains remain incompletely understood. In the present study, we sought to investigate how the differential metabolomes can be used to distinguish UPEC from non-UPEC strains and to determine the associated regulatory roles of siderophore biosynthesis. Our results are the first to demonstrate that the identified differential metabolomes strongly differentiated UPEC from non-UPEC strains. Furthermore, we performed metabolome assays of mutants with different patterns of siderophore deletions; the data revealed that the mutations of all four siderophores exerted a stronger modulatory role on the differential metabolomes of the UPEC and non-UPEC strains relative to the mutation of any single siderophore and that this modulatory role primarily involved amino acid metabolism, oxidative phosphorylation in the carbon fixation pathway, and purine and pyrimidine metabolism. Surprisingly, the modulatory roles were strongly dependent on the type and number of mutated siderophores. Taken together, these results demonstrated that siderophore biosynthesis coordinately modulated the differential metabolomes and thus may indicate novel targets for virulence-based diagnosis, therapeutics, and drug development related to urinary tract infections

Correlation of VSTM1-v2 expression level and IL-17A mRNA expression level.

<p>(A) The expression IL-17A mRNA was detected with real time-PCR. Relative IL-17A mRNA was normalized to GAPDH in PBMCs from patients with rheumatoid arthritis (RA; n = 40) and healthy controls (HC; n = 40). Data are shown as scatter plot. <i>P</i> value was determined by Mann-Whitney test. (B) The correlation between VSTM1-v2 mRNA expression level and IL-17A mRNA expression level was determined by using Spearman test. The relation between VSTM1-v2 and IL-17A was presented.</p