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1.
Chinese Journal of Dermatology ; (12): 310-313, 2019.
Artigo em Chinês | WPRIM | ID: wpr-745786

RESUMO

Objective To establish a culture method for primary human nail matrix cells in serumfree media.Methods Nail matrix tissues were collected from 9 patients,who received nail or toe amputation and nail bed repair in Peking University Shenzhen Hospital between January 2016 and December 2016,and cultured in the serum-free DEME/F-12 media at a 37℃ incubator with an atmosphere of 5% CO2 in air for 2-3 days.Then,primary human nail matrix cells were cultured in keratinocyte serumfree media (CnT-07),and the morphology of human nail matrix cells was observed by microscopy during the culture process.Immunofluorescence cytochemistry with anti-keratin 5 (K5) and K10 was performed to identify the acquired cells,and flow cytometry to analyze the cell purity.Results After 2 or 3 days of the culture,some cells began to crawl out from the tissue.On day 10,large cell masses were formed,some cells were morphologically similar to epithelioid cells arranged in a paving stone-like pattern,and some were flat giving a spindle-shaped or star-shaped appearance.Immunofluorescence cytochemistry showed that some cells could express both K5 and K10,which proved the existence of nail matrix cells,and 37.6% of the cells expressed K10.Conclusion Human primary nail matrix cells could be successfully cultured by using the tissue culture method with serum-free culture media,and the nail matrix cells cultured in vitro can express both K5 and K10.

2.
Chinese Journal of Medical Aesthetics and Cosmetology ; (6): 330-334, 2019.
Artigo em Chinês | WPRIM | ID: wpr-756581

RESUMO

Objective To investigate the wound-healing process in a rat model of skin full-thickness incisions and to detect related possible mechanism.Methods Twenty-four female rats were selected and the dorsal skin of rats was used as the experimental area.A cutaneous excision (6 mm diameter) was made on the back of each animal,close to the cervical area.The dorsal skin of every rat was allocated to three groups which were treated with physiological saline,human recombinant epidermal growth factor (rhEGF),and CDPs,respectively.After making a rat model of skin incisions,we observed the wound healing process,took photos of the wounds under a digital microscope,and use sulfuric graph paper to record the size of every wound.At the 3rd,6th,9th,12th day after modeling,6 rats were killed,and mRNA expression of K10,K14,and EGF was detected in the skin tissues using a RT-PCR technique.Results At the 6th and 12th day after modeling,there were significant differences between the experimental group and the blank control group (P<0.05).The gene expression of EGF,K-14 in the third day and that of EGF,K-10 and K-14 in the 6th and 12th day were upregulated compared with control group,and there were significant differences between them (P < 0.05).Conclusions CDPs have a beneficial effect on the acceleration of skin wound healing,possibly due to increasing keratinocyte proliferation and up-regulating the expression of K10,K14 and EGF genes.

3.
Chinese Journal of Dermatology ; (12): 186-188, 2018.
Artigo em Chinês | WPRIM | ID: wpr-710355

RESUMO

Objective To identify mutations in keratin genes (KRT1 and KRT10) in a pair of twins with bullous congenital ichthyosiform erythroderma (BCIE),and to explore the relationship between the causative genes and phenotypes.Methods Clinical data were collected from a pair of twins with BCIE and their family members.Peripheral blood samples were obtained from the twins,their old brother and parents,and DNA was extracted from these blood samples.Polymerase chain reaction (PCR)was performed to amplify all the coding exons and their flanking sequences of the KRT1 and KRT10 genes,and 100 unrelated healthy persons served as controls.Results The 11-year-old male proband presented with recurrent blisters,hypertrophy and desquamation all over the body for 11 years.His twin brother had similar skin lesions.Skin examination of the proband showed diffuse erythema covered with thick scaly crusts on the trunk and extremities.Blisters,bullae and erosions due to ruptured blisters were observed locally with tenderness on palpation.There were obvious hyperkeratotic and hard lesions on the big joints of the extremities.Diffuse hyperkeratosis could be seen on the palms and soles.A mutation c.591 + 1G > A was identified at position 1 in intron 1 of the KRT1 gene in the twins,but not in the 3 healthy family members or the 100 unrelated healthy controls.Conclusion The mutation c.591 + 1G > A at position 1 in intron 1 of the KRT1 gene may contribute to the clinical phenotype of the twins with BCIE.

4.
Chinese Journal of Endemiology ; (6): 365-368, 2012.
Artigo em Chinês | WPRIM | ID: wpr-642766

RESUMO

Objective To observe the influences of different doses of sodium arsenite on mRNA transcription of keratinizing related and nuclear factor E2-related factor 2(Nrf2) genes in HaCaT cells.Methods Cell proliferation was evaluated by Cell Counting Kit-8(CCK-8) assay after the HaCaT cells were exposed to 0.00,3.13,6.25,12.50,25.00,50.00,75.00,100.00 μ mol/L sodium arsenite for 48 h,respectively.Based on the previous results of cell proliferation,0.00(control),6.25,12.50,and 25.00 μmol/L of sodium arsenite were selected to treat HaCaT cells for 48 h,respectively.The mRNA expression of keratin 1,keratin 10,involucrin,loricrin and Nrf2 were detected by real-time fluorescent quantitative PCR.ResultsCompared with the control group (100.05%),HaCaT cell proliferation rates(83.06%,51.04%,39.52%,24.51%,16.99% and 9.04%) were significantly lower in 6.25,12.50,25.00,50.00,75.00 and 100.00 μ mol/L of sodium arsenite groups and the 50% inhibiting concentration was 12.38 μmol/L.Compared with the control group( 1.06 ± 0.28,1.00 ± 0.12,1.00 ± 0.08),the mRNA expression of keratin 1,involucrin and loricrin (0.08 ± 0.04,0.13 ± 0.12,0.05 ± 0.03;0.47 ± 0.11,0.21 ± 0.09,0.10 ± 0.15; 0.50 ± 0.27,0.31 ± 0.10,0.57 ± 0.23) were significantly decreased(all P < 0.05) in HaCaT cells treated with 6.25,12.50,25.00 μmol/L sodium arsenite,respectively.But keratin 10 mRNA expression showed a rise trend and the 6.25 μmoL/L sodium arsenite group (1.83 ± 0.45) was significantly higher than that of the control( 1.07 ± 0.14,P < 0.05 ).The Nrf2 mRNA expressions of HaCaT cells in 12.50,25.00 μmol/L sodium arsenite groups(0.13 ± 0.07,0.69 ± 0.33) were significantly lower than that of the control ( 1.00 ± 0.09,all P < 0.05 ).ConclusionsThe cellular proliferation and keratinization are decreased when HaCaT cells are exposed to sodium arsenite,which may be regulated by lowering Nrf2 mRNA transcription.

5.
Chinese Journal of Dermatology ; (12): 704-707, 2012.
Artigo em Chinês | WPRIM | ID: wpr-420897

RESUMO

Objective To identify gene mutations in two families with epidermolytic hyperkeratosis (EHK).Methods Clinical data were collected from two families with EHK.Peripheral blood was isolated from the probands and unaffected family members in the families as well as from 50 healthy controls.PCR was performed to amplify the encoding exons and flanking intron regions of KRT1 and KRT10 genes followed by direct DNA sequencing.Results Two mutations in the KRT10 gene,including a heterozygous acceptor splice site mutation in intron 4 (c.1030-2 A>G) and a heterozygous missense mutation c.467 G>A,were identified in the probands of both families,but absent in the unaffected family members or healthy controls.ConclusionThe splice site mutation c.1030-2 A>G and missense mutation c.467 G>A might be responsible for the phenotype of EHK in the two families.

6.
Chinese Journal of Endemiology ; (6): 162-164, 2009.
Artigo em Chinês | WPRIM | ID: wpr-642236

RESUMO

Objective To observe expression of the keratin 10 (K10), c-myc and cox2 in buccal mucous membrane exfoliated cells, and changes of liver function of coal-burning arsenism patients, in order to explorate coal-burning arsenic poison circumference biology symbol. Methods The buccal mucous membrane exfoliated cells were collected from both arsenism patients and normal controls. Real-time PCR was employed to detect the changes of K10, c-myc, cox2 genes expression in these cells. Simultaneously, circumference venous blood was extracted and examinated liver function. Results K10 was found obviously overexpressed in arsenism patients(3.60±0.94) compared to control(1.82±0.68), the difference had statistics significance(t=2.15, P<0.05), c-mye and cox2 did not found obviously changed(c-myc: 3.50±2.77,3.39±2.07; cox2:5.90±1.40,4.73±1.91; t=1.26,1.65, P> 0.05). The serum ALT of patients(25.83±2A5) obviously increased than control(36.86±1735, t=2.55, P<0.05). Conclusions Expression of K10 gene in buccal mucous membrane cells may be regarded as sensitive molecular markers for skin pathologic changes in arsenic patients. The liver is a sensitive target organ of inorganic arsenic.

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