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Background@#Previous studies demonstrate that eccrine sweat glands are innervated by both cholinergic and adrenergic nerves. However, it is still unknown whether the secretory coils and ducts of eccrine sweat glands are equally innervated by the sympathetic nerve fibers. To well understand the mechanisms on sweat secretion and reabsorption, the differential innervation of secretory coils and ducts in human eccrine sweat glands was investigated in the study.@*Methods@#From June 2016 to June 2017, six human skins were fixed, paraffin-embedded, and cut into 5 μm-thick sections, followed by costaining for nerve fiber markers protein gene product 9.5 (PGP 9.5), tyrosine hydroxylase (TH) and vasoactive intestinal peptide (VIP), and eccrine sweat gland markers K7, S100P, and K14 by combining standard immunofluorescence with tyramide signal amplification (IF-TSA). Stained sections were observed under the microscope, photographed, and analyzed.@*Results@#The fluorescent signals of PGP 9.5, TH, and VIP were easily visualized, by IF-TSA, as circular patterns surrounding eccrine sweat glands, but only PGP 9.5 could be observed by standard IF. The IF-TSA method is more sensitivity than standard IF in detecting antigens expressed at low levels. PGP 9.5, TH, and VIP appeared primarily surrounding the secretory coils and sparsely surrounding the sweat ducts.@*Conclusion@#Sweat secretion is mainly controlled by autonomic nerves whereas sweat reabsorption is less affected by nerve activity.
Subject(s)
Humans , Eccrine Glands , Fluorescent Antibody Technique , Nerve Fibers , Sweat Glands , Vasoactive Intestinal PeptideABSTRACT
Background and purpose:Adenosine triphosphate-binding cassette superfamily G member 2 (ABCG2), which has been found over-expressed in a variety of cancer cells, takes part in the drug resistance of cancer through effux of anticancer drugs. The purpose of this study was to investigate the mechanisms of human glioblastoma cells sensitivity to pyropheophorbide-a methyl ester (MPPa)-mediated photodynamic therapy (PDT) eradicating tumour cells and its relationship to ABCG2.Methods:U87 and A172 glioma cell lines in the logarithmic growth phase were selected and exposed to the treatment of MPPa-PDT and MPPa-PDT+fumitremorgin C (FTC) respectively. The cell viability was measured with the use of CCK-8 assay. The expression of ABCG2 was detected by Western blot. The intracellular contents of MPPa in each group without illumination were tested by lfow cytometry. Flow cytometry with AnnexinⅤ-FITC/PI double staining was used to detect the cell apoptotic rate. DCFH-DA staining was used to assess the generation of intracellular reactive oxygen species (ROS).Results:The MPPa-mediated PDT could eradicate A172 and U87 cancer cells in an energy-dependent manner. The light energy density in A172 was 8 times of that in U87 when the cell viability reached median lethal dose after MPPa-mediated PDT. The high expression of ABCG2 in A172 cells affected the accumulation of intracellular MPPa. Inhibition of ABCG2, not only could enhance the eradicating effect of MPPa-PDT on A172 cells, but also could increase the yield of ROS triggered by MPPa-PDT and the accumulation of intracellular MPPa.Conclusion:The human glioblastoma cell line A172 is insensitive to MPPa-mediated PDT. The mechanism may relate to ABCG2, which decreases the MPPa content in cancer cells through effux of MPPa, resulting in decline of cytotoxicity.
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Objective To investigate the mechanism by which methyl pyropheophorbide-a-mediated photodynamic therapy (Mppa-PDT) inhibit cell viability and induce apoptosis in human ovarian cancer cell line SKOV3. Methods Human ovarian cancer cells SKOV3 at the logarithmic growth phase were divided into Mppa-PDT treated group (both Mppa and PDT treated group) and control groups (the blank group, the only Mppa treated group and only PDT treated group). After Mppa-PDT treatment, the cell viability was examined with CCK-8 assay; cell apoptosis was detected by flow cytometry with Annexin -FITC/PI; and nuclear morphological changes during cell apoptosis was detected by DAPI staning. Moreover, the celluar reactive oxygen species (ROS) were detected by DCFH-DA staining; DNA damage was observed by single cell gel electrophoresis; and the protein expression of p53, Caspase-3, Bax, and Bcl-2 were assessed by Western blotting analysis. Results (1) Mppa-PDT could greatly suppress the cell viability of human ovarian cancer cells SKOV3 in a dose-dependent manner. (2) The cell apoptosis rate of Mppa-PDT treated group was significanlty higher than those of three control groups (blank group, Mppa group and PDT group) (P0.05). (3) After treating with Mppa-PDT, DAPI staining showed strongly stained nuclei of the apoptotic cells; DCFH-DA staining displayed higher level of ROS than those of the three control groups; single cell gel electrophoresis showed greater DNA damage than those of the three control groups; and Western blotting analysis showed that the expression of p53, Caspase-3 and Bax protein was increased and Bcl-2 protein was decreased (P<.05). Conclusion Mppa-PDT can significantly suppress cell viability and induce apoptosis in human ovarian cancer cell SKOV3, accompanied by DNA damage and the activation of mitochondrial apoptosis pathway.
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<p><b>OBJECTIVE</b>To construct and identify the stable expression system of NIH3T3 fibroblast with eukaryotic expression vector of human transforming growth factor beta3 (pcDNA3.1 (-)/TGFbeta3). So as to investigate the proliferation of NIH3T3 fibroblasts transfected with hTGFbeta3 gene stably.</p><p><b>METHODS</b>The stable transfection of NIH3T3 fibroblasts with recombinant plasmid expressing hTGFbeta3 was established by using LipofectamineTM2000 and G418 selection. The mRNA and protein expression of TGFbeta3 were detected by the RT-PCR and Western blot method, respectively. Microscope and MTT were adopted to examine the proliferation of the stable expression system of fibroblasts with hTGFbeta3.</p><p><b>RESULTS</b>After G418 selection, RT-PCR and Western blot analysis, 7 out of 10 cell lines transfected with pcDNA3.1 (-)/TGFbeta3 expressed with very high level of TGFbeta3, as compared with vector control transfectants that showed no expression, and compared with the other cell lines that expressed relatively low level. The stable transfection of NIH3T3 fibroblasts growth slowed down significantly (P < 0.05).</p><p><b>CONCLUSION</b>The stable expression system of NIH3T3 fibroblast with hTGFbeta3 were constructed successfully. The TGFbeta3 gene could inhibit the proliferation of NIH3T3 fibroblasts in vitro.</p>
Subject(s)
Animals , Humans , Mice , Cell Proliferation , Fibroblasts , Metabolism , NIH 3T3 Cells , Plasmids , Transfection , Transforming Growth Factor beta3 , Genetics , MetabolismABSTRACT
<p><b>AIM</b>To explore the causes of the difference in spermatogenic suppression between responders and non-responders in Chinese men treated with levonorgestrel (LNG) implants plus testosterone undecanoate (TU) injectable.</p><p><b>METHODS</b>The 16 Chinese volunteers treated were divided into two groups in regard to the sperm count during the treatment period, 7 men in the responder group (Group R), including 6 azoospermia and one severe oligozoospermia, and the remaining 9 in the non-responder group (Group N), including 4 oligozoospermia and 5 with sperm counts greater than 20 x 10(6)/mL. The differences in serum profiles of FSH, LH, T, LNG and T/LH ratio were compared between the two groups and the correlation between the seminal fluid parameters and serum reproductive hormones was analyzed.</p><p><b>RESULTS</b>The serum FSH level was lower in Group R than that in Group N (P<0.05), while the serum LH and LNG levels were higher in Group R than those in Group N (P<0.05). The sperm density (P<0.01, r=0.235), motility(P<0.01, r=0.326) and vitality (P<0.01, r=0.219) showed significantly positive correlation with the serum FSH level.</p><p><b>CONCLUSION</b>The blood LNG and T levels, the degree of FSH inhibition and/or the sensitivity of the pituitary-testis axis to exogenous steroids, as well as the individual spermatogenetic potential and the functional status of the Leydig cells may be factors bringing about individual differences in spermatogenic suppression in Chinese men treated with LNG and TU.</p>