Characterization of primary endometrial epithelial cells (EECs) and endometrial stromal cells (ESCs).

<p>The cobblestone structure and expression of cytokeratin were shown in EECs (a, b) and the spindle-shape structure and vimentin expression were shown in ESCs (e, f). The negative control shown of cytokeratin and vimentin involved use of mouse IgG in place of primary antibody (c, g). Western blot analysis of estrogen receptor (ER) and progesterone receptor (PR) protein expression were shown separately in the endometrial cells (d, h). Results were quantified by densitometry analysis of the bands and normalization to β-actin protein. Data represent the mean ± standard error of the mean (SEM) from five independent experiments. Columns with different superscript letters are significantly different (<i>P</i> < 0.05). Scale bar = 30 µm.</p

Different developmental stages of goat embryos produced <i>in vitro</i>.

<p>Transferred IVF blastocysts (a) or SCNT blastocysts (b) showed clear inner cell masses (ICMs) and trphoectodermal cells (TE) under bright field at Day 7. Both the IVF blastocysts (c) and SCNT blastocysts (d) attached to the endometrial epithelial cells after co-cultured for 24 h. The trophoblast of IVF (e) and SCNT (f) underwent outgrowth on the epithelial cells monolayer after co-cultured for 72 h. Scale bar = 50 µm.</p

The secretion and expression of cytokines by uterine DBA⁺ leukocytes in response to embryos and steroids.

<p>ELISA analyses of the concentrations of IFN-γ (a) and VEGF (c) secreted by uterine DBA⁺ leukocytes in response to cloned and fertilized embryos. Representative western blotting and densitometric analysis of IFN-γ (b) and VEGF (d) normalized to β-actin to correct for protein loading. <i>Columns</i> and <i>vertical bars</i> represent the mean ± SEM from at least five independent experiments, and analyzed by two-way ANOVA with treatment group and/or embryos as the independent variable(s). Columns with different superscript letters are significantly different from each other (<i>P</i> < 0.05).</p

Flow cytometry analysis of the expression of CD56⁺CD16^- on goat uterine leukocytes.

<p>Representative flow cytometry analysis of the percentage of CD56⁺CD16^- cells on isolated goat uterine leukocytes in response to cloned and fertilized embryos (a). The absolute numbers of uterine CD56⁺CD16^- leukocytes over 120-hr incubation with different conditioned mediums or control medium (b). Fold expansion of total numbers of isolated goat uterine leukocytes after 120-hr incubation with different conditioned mediums or control medium in comparison to the cells at the beginning of the culture period (c). <i>Columns</i> and <i>vertical bars</i> represent the mean ± SEM from at least five independent experiments, and analyzed by two-way ANOVA with treatment group and/or embryos as the independent variable(s). Columns with different superscript letters are significantly different from each other (<i>P</i> < 0.05).</p

Isolation and identification of uterine <i>Dolichos biflorus</i> agglutinin (DBA) positive leukocytes in non-pregnant goat endometrium.

<p>Lymphocytes were identified by their size and granularity in the FSC/SSC dot plot. Representative flow cytometry analysis of the percentages of DBA⁺ and CD56⁺ cells among CD45⁺ leukocytes in the non-pregnant goat endometrium (a). Differential interference contrast (DIC) image of viable goat uterine leukocytes attached to DBA⁺ lectin-coated magnetic beads (b) and detached after addition of 0.1 M Gal-NAc (c). The expression of CD56, CD16 on the isolated uterine leukocytes was evaluated by flow cytometer (d). Representative microphotographs for DBA lectin reactive uterine leukocytes scattered between endometrial glands (e) and stroma (f) of the non-pregnant goat endometrium. No positive reaction was seen in the control reaction performed with DBA lectin inhibited with NacGal (g). Representative flow cytometry analysis of the expression of perforin and Granzyme B on gated CD56⁺DBA⁺ leukocytes isolated (h). Scale bar = 30 µm (b, c) and 90 µm (e-g).</p

Dual-Modal Imaging-Guided Theranostic Nanocarriers Based on Indocyanine Green and mTOR Inhibitor Rapamycin

The development of treatment protocols that resulted in a complete response to photothermal therapy (PTT) was usually hampered by uneven heat distribution and low effectiveness. Here, we reported an NIR fluorescence and photoacoustic dual-modal imaging-guided active targeted thermal sensitive liposomes (TSLs) based on the photothermal therapy agent Indocyanine green (ICG) and antiangiogenesis agent Rapamycin (RAPA) to realize enhanced therapeutic and diagnostic functions. As expected, the <i>in vitro</i> drug release studies exhibited the satisfactory result of drug released from the TSLs under hyperthermia conditions induced by NIR stimulation. The <i>in vitro</i> cellular studies confirmed that the FA-ICG/RAPA-TSLs plus NIR laser exhibited efficient drug accumulation and cytotoxicity in tumor cells and epithelial cells. After 24 h intravenous injection of FA-ICG/RAPA-TSLs, the margins of tumor and normal tissue were accurately identified via the <i>in vivo</i> NIR fluorescence and photoacoustic dual-modal imaging. In addition, FA-ICG/RAPA-TSLs combined with NIR irradiation treated tumor-bearing nude mice inhibited tumor growth to a great extent and possessed much lower side effects to normal organs. All detailed evidence suggested that the theranostic TSLs which were capable of enhancing the therapeutic index might be a suitable drug delivery system for dual-modal imaging-guided therapeutic tools for diagnostics as well as the treatment of tumors