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1.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 369-378, 2023.
Article in Chinese | WPRIM | ID: wpr-973232

ABSTRACT

ObjectiveTo compare the therapeutical effect of exosomes derived from fibroblasts and mesenchymal stem cells on acute wound healing. MethodsPrimary human dermal fibroblasts (hDF) were isolated, cultured and identified. Human bone marrow mesenchymal stem cell exosomes (hMSC-EXO) and hDF exosomes (hDF-EXO) were extracted by ultracentrifuga tion. After 24 h of coincubation with hDF-EXO or hMSC-EXO, hDFs proliferation and migratory capacity were evaluated by cell counting kit-8 (CCK8) assay and scratch test. Full-thickness cutaneous wounds were created on 8-week-old female C57BL/6 mice, and topically applied with PBS (control), hDF-EXO or hMSC-EXO. Wounds were measured at day 0, 2, 4, 7, and the uptake of exosomes in wound was observed at day 1. Quantitative PCR (qPCR) analysis was performed to detect the mRNA expression levels of TNF-α, IL-6, IL-1β, IL-10 in wound at day 1. HE staining was conducted to analyze the histological structure of wounds at day 7, while immunofluorescence staining was used to examine expression of PDGFR-α、α-SMA、Ki67. ResultshDF exhibited certain fibrolast-like characteristics with respect to expression of cell surface markers and specific proteins. hDF-EXO and hMSC-EXO presented exosomal morphology, size, and markers, and both concentrations were not statistically different (P>0.05); CCK8 assay showed that both exosomes promoted hDF cell viability, compared with the negative control (P<0.01), and hDF-EXO group had greater cell viability than hMSC-EXO group (P<0.01). Scratch test indicated that hDF-EXO induced a significant increase in scratch healing rate versus the negative control (P<0.01), hMSC-EXO (P<0.05). In vivo experiments showed wound tissues took up exosomes at day 1. qPCR detected TNF-α, IL-6, IL-1β expression levels in wound at day 1 were lower in exosomes group than in the control group, and were the lowest in hMSC-EXO group (all P<0.01). Wound areas were measured smaller at day 7 in exosomes group than in the control group (all P<0.01) and hDF-EXO group had better closure than hMSC-EXO group (P<0.05). HE staining revealed that compared with control group, scar, incomplete epidermis and few collagen deposition remained in the hMSC-EXO group, whereas hDF-EXO group showed re-epithelialization, continuous neo-epidermis and regenerated dermis. Immunofluorescence staining suggested that the number of fibroblasts, myofibroblasts, proliferating cells was higher in both exosomes group than that in the control group, especially the highest in hDF-EXO group. ConclusionOur study shows both exosomes accelerate wound healing, whereas hDF-EXO is more effective in promoting fibroblasts proliferation, migration, transition to myofibroblasts, and hMSC-EXO may play a role in inhibiting inflammatory reaction during early stage of wound healing.

2.
National Journal of Andrology ; (12): 392-397, 2013.
Article in Chinese | WPRIM | ID: wpr-350893

ABSTRACT

<p><b>OBJECTIVE</b>To explore the P75NTR expression in the mouse testis and its relationship with nestin.</p><p><b>METHODS</b>We observed the location of the expressions of P75NTR and nestin in the testis of the nestin-GFP transgenic mouse on postnatal day (PND) 5, 14 and 30 using immunofluorescence, and detected the expression levels of P75NTR in the testicular tissue of mice in different age groups by real-time quantitative PCR (RTqPCR) and flow cytometry. Then we cultured the P75NTR positive cells in neural stem cell culture medium and observed their neuronal differentiation capacity by orientation differentiation.</p><p><b>RESULTS</b>Immunofluorescence showed the expressions of P75NTR and nestin in the Leydig cells of the mouse testis. RTqPCR and flow cytometry exhibited the peak of the P75NTR expression on PND 14. The positive rates of P75NTR were (2.88 +/- 0.52), (9.54 +/- 1.81) and (2.63 +/- 0.43)% on PND 5, 14 and 30, respectively. The P75NTR positive cells obtained also expressed nestin and P75NTR and had the capacity of neuronal differentiation.</p><p><b>CONCLUSION</b>P75NTR and nestin are co-expressed in the Leydig cells of the mouse testis, and the P75NTR positive cells have the ability of neural differentiation, which is presumably attributed to neural crest cells.</p>


Subject(s)
Animals , Male , Mice , Intermediate Filament Proteins , Genetics , Metabolism , Leydig Cells , Metabolism , Mice, Inbred C57BL , Mice, Transgenic , Nerve Tissue Proteins , Genetics , Metabolism , Nestin , Receptor, Nerve Growth Factor , Genetics , Metabolism , Testis , Cell Biology , Metabolism
3.
Journal of Southern Medical University ; (12): 716-719, 2010.
Article in Chinese | WPRIM | ID: wpr-355036

ABSTRACT

<p><b>OBJECTIVE</b>To observe the expression of protein arginine N-methyltransferase (PRMT) genes in the lung and spleen of E3 rats with acute asthma.</p><p><b>METHODS</b>E3 rats with ovalbumin-induced pulmonary inflammation were divided into two groups (n=10), and the validity of the acute asthma model was evaluated by histological observation with HE and PAS staining and by measurement of NO production. Semi-quantitative RT-PCR was employed to detect the expressions of PRMT1-PRMT6 genes in the lung and spleen tissues of the rats.</p><p><b>RESULTS</b>In the lung tissue of the asthmatic rats, the gene expressions of PRMT1 (P<0.01), PRMT2 (P<0.01), PRMT3 (P<0.05) and PRMT5 (P<0.05) were significantly increased, but the expression of PRMT4 gene (P<0.05) was significantly decreased as compared with those in the control tissue. In the spleen tissue of the asthmatic rats, the expressions of PRMT2 (P<0.05) and PRMT5 genes (P<0.05) showed a significant increase as compared with those in the control rat tissue.</p><p><b>CONCLUSION</b>The gene expressions of PRMTs vary significantly between asthmatic rats and control rats, suggesting that PRMTs play an important role in the post-translational modification process of asthma-related genes.</p>


Subject(s)
Animals , Female , Male , Rats , Acute Disease , Asthma , Protein Processing, Post-Translational , Protein-Arginine N-Methyltransferases , Classification , Genetics , Metabolism , Random Allocation , Rats, Inbred Strains
4.
Journal of Korean Society of Endocrinology ; : 398-405, 2004.
Article in Korean | WPRIM | ID: wpr-131914

ABSTRACT

The familial occurrence of a pituitary adenoma associated with multiple endocrine neoplasia (MEN) type 1 or Carney complex is a well-recognized entity. However, an isolated familial somatotropinoma is a rare inherited disease, which is characterized by clustering of a somatotrophic adenoma and acromegaly or gigantism in a family, but without other manifestations of MEN type 1, with only 68 cases, in 28 families, described in the literature. The mode of inheritance is autosomal dominant, with incomplete penetration, but the genetic background of these pituitary adenomas remains unknown. A family exists where both the father and son were affected. Endocrinological investigations confirmed hypersecretion of GH and IGF-1, and the pituitary adenomas were identified by magnetic resonance image in both cases. There was no symptom of MEN type 1 or other form of endocrine dysfunction. Herein is reported a case of an isolated familial somatotropinoma in Korea, with a review of the literature


Subject(s)
Humans , Male , Acromegaly , Adenoma , Carney Complex , Fathers , Gigantism , Growth Hormone-Secreting Pituitary Adenoma , Insulin-Like Growth Factor I , Korea , Multiple Endocrine Neoplasia , Pituitary Neoplasms , Somatotrophs , Wills
5.
Journal of Korean Society of Endocrinology ; : 398-405, 2004.
Article in Korean | WPRIM | ID: wpr-131911

ABSTRACT

The familial occurrence of a pituitary adenoma associated with multiple endocrine neoplasia (MEN) type 1 or Carney complex is a well-recognized entity. However, an isolated familial somatotropinoma is a rare inherited disease, which is characterized by clustering of a somatotrophic adenoma and acromegaly or gigantism in a family, but without other manifestations of MEN type 1, with only 68 cases, in 28 families, described in the literature. The mode of inheritance is autosomal dominant, with incomplete penetration, but the genetic background of these pituitary adenomas remains unknown. A family exists where both the father and son were affected. Endocrinological investigations confirmed hypersecretion of GH and IGF-1, and the pituitary adenomas were identified by magnetic resonance image in both cases. There was no symptom of MEN type 1 or other form of endocrine dysfunction. Herein is reported a case of an isolated familial somatotropinoma in Korea, with a review of the literature


Subject(s)
Humans , Male , Acromegaly , Adenoma , Carney Complex , Fathers , Gigantism , Growth Hormone-Secreting Pituitary Adenoma , Insulin-Like Growth Factor I , Korea , Multiple Endocrine Neoplasia , Pituitary Neoplasms , Somatotrophs , Wills
6.
Chinese Journal of Medical Genetics ; (6): 19-22, 2003.
Article in Chinese | WPRIM | ID: wpr-248510

ABSTRACT

<p><b>OBJECTIVE</b>To determine the molecular basis of late onset ornithine transcarbamylase (OTC) deficiency in a Chinese family of Han nationality and the exon sequences of OTC gene of this patient.</p><p><b>METHODS</b>Polymerase chain reaction-single strand conformation polymorphism and direct sequencing were used to identify the mutation type.</p><p><b>RESULTS</b>A missense mutation E122G in the conserved residue of exon 4 was identified which is unreported before.</p><p><b>CONCLUSION</b>The E122G mutation in human OTC gene may cause late onset OTC deficiency.</p>


Subject(s)
Child, Preschool , Female , Humans , Male , Age of Onset , Base Sequence , DNA , Chemistry , Genetics , DNA Mutational Analysis , Family Health , Fatal Outcome , Models, Molecular , Mutation, Missense , Ornithine Carbamoyltransferase , Chemistry , Genetics , Ornithine Carbamoyltransferase Deficiency Disease , Genetics , Pathology , Pedigree , Polymorphism, Single-Stranded Conformational , Protein Structure, Secondary
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