A novel pathogenic mutation in the NEMO gene in a family with incontinentia pigmenti / 中华皮肤科杂志

Objective:To identify gene mutations in a family with incontinentia pigmenti, in order to confirm pathogenic mutations.Methods:Clinical data were collected from all patients in a family with incontinentia pigmenti. DNA was extracted from peripheral blood samples obtained from the patients, healthy members in the family, and 100 unrelated healthy controls, and Sanger sequencing was performed for all exons and their flanking sequences of the NEMO gene.Results:Totally, there were 4 patients in the 4-generation family, who all presented with typical skin lesions and different symptoms. Genetic testing indicated that the proband and the other 3 patients all carried a heterozygous nonsense mutation c.1153C>T (p.Gln385X) at position 1153 in exon 8 of the NEMO gene, which led to the substitution of the glutamine codon (CAG) by the termination codon (TAG) at amino acid position 385. The mutation was not identified in the 14 healthy relatives or 100 unrelated healthy controls. The mutation cosegregated with incontinentia pigmenti in the family. Database searching confirmed the mutation to be a novel nonsense mutation, and it was considered as a very strong pathogenic locus according to the American College of Medical Genetic and Genomics guidelines.Conclusion:The mutation c.1153C>T in the NEMO gene is associated with the occurrence of incontinentia pigmenti in this family.

Application of radiochemical separation and a-spectrometry in individual dose monitoring for occupational plutonium internal exposure / 中华放射医学与防护杂志

Objective To explore the applicable conditions for using urine plutonium monitoring data to assess personal internal doses,in order to provide references for the occupational health management and the urine plutonium monitoring in nuclear sector.Methods Using some plutonium mixtures from DOE nuclear facilities,as an example,the urine plutonium levels were estimated through simulation calculation at 1 mSv effective dose arising from either acute or chronic inhalation of plutonium compounds,respectively.The results were compared with the typical detection limit of radiochemical separation and α-spectrometry.The feasibility of urine plutonium monitoring for dose assessment of internal radiation exposure was discussed.Results Only for type M plutonium compunds,1 mSv detection limit can be achieved using radiochemical separation and α-spectrometry within 10 d after inhalation.Conclusions Before the monitoring plan of urine plutonium is made,detection limits of monitoring method should be considered.Internal dose could be accessed using workplace air monitoring and working hours when necessary.

Construction of tissue-engineered skin by culture of human amniotic epithelial cells on human deepidermized dermis for the repair of skin defects in nude mice: an experimental study / 中华皮肤科杂志

Objective To evaluate the feasibility of repair of full-thickness skin defects in nude mice with tissue-engineered skin which was constructed by culture of human amniotic epithelial cells (hAECs) and fibroblasts on human de-epidermized dermis (DED).Methods Healthy human amniotic tissues were treated with trypsin at a low concentration in multi-steps to prepare hAECs,and a two-step collagenase digestion was used to treat healthy children's prepuce tissues to prepare fibroblast suspensions.When fibroblasts were cultured in vitro up to passage 3-5 and hAECs up to passage 2,they were seeded on the reticular dermal surface and basement membrane surface of the DED respectively to construct the tissueengineered skin.A total of 20 heahhy male nude mice aged 3-4 weeks were enrolled into this experiment,and full-thickness skin defects were made on the middle of the back of mice.Then,these mice were randomly divided into 2 groups by using a lottery method,and reconstructed full-thickness tissueengineered skin grafts and vaseline oil gauze were used to cover the wounds in the tissue-engineered skin group and control group respectively.The whole body and transplantation sites of the nude mice were observed on day 7,14,21 and 28 after transplantation,the wound healing time and rate were compared between the above two groups,and skin tissues at the transplantation site were harvested at 4 weeks after transplantation and subjected to histological examination.Results HAECs had stem-cell characteristics and expressed octamer-binding protein-4 (OCT-4) and embryonic marker stage-specific embryonic antigen4 (SSEA-4).After 2-week organ culture,the in vitro reconstructed tissue-engineered skin showed 4-9 continuous layers of stratified epithelium,and the histological structure of the epidermis was similar to that of the normal human skin.Compared with the control group,the tissue-engineered skin group showed significantly higher wound healing rates on day 7,14 and 21 after transplantation (57.49％ ± 6.11％ vs.22.93％ ± 4.26％,92.80％ ± 3.10％ vs.54.57％ ± 7.94％,98.83％ ± 0.25％ vs.91.16％ ± 4.79％,respectively;n =10,t =27.36,32.23,11.80,respectively,all P ＜ 0.001),shorter wound healing time [(21.51 ± 1.51) d vs.(28.80 ± 1.14) d,n =10,t =42.23,P ＜ 0.001],with the color of skin grafts closer to that of autologous skin on day 28 after transplantation.Histological examination revealed distinct stratification of the epithelium,obvious keratinization and favorable growth of cells in the dermis in the tissue-engineered skin group,but thin epithelium with some defects,indistinct stratification of the dermis,and inflammatory cell infiltration in the control group.Condusion Tissue-engineered skin constructed by the culture of hAECs and fibroblasts on human DED can survive in nude mice after transplantation,resulting in a more favorable healing of wounds,and is expected to serve as a kind of ideal tissue-engineered skin.

Association study of alopecia areata susceptibility loci in Han Chinese population / 安徽医科大学学报

Objective To study the correlation between alopecia areata and the susceptibility genes identified in our previous study with Han Chinese population. Methods The study performed an independent replication study using 736 cases and 1 840 controls. DNA was extracted from peripheral blood leukocytes by salting out with saturat-ed NaCl solution according to standard methods. The evidence for association had been obtained from former gene study. Then a fine-mapping study and genotyped the locus with an additional 17 SNPs were performed. Data were analyzed with the use of Plink 1. 07 software. Results Only one SNP achieved nominal significance, rs3087243 (P = 0. 041, OR = 1. 18, 95% CI = 1. 01 ~ 1. 38). The other 16 genes (TLR1, DMBT1, CHIT1, GBP4, CIITA, IL31RA, CD96, INPPL1, MASP2, IL-13, KIAA0350, PTPN22, SPATA5, TRAF1 / C5, IL1A, IL2R) failed. A further stratification analysis of alopecia areata was adopted, including the analysis of family history, the age of on-set and the severity of alopecia areata. The stratification analysis revealed that the age of onset > 20 years achieved nominal significance P < 0. 05 for one SNP, rs2416808 (P = 0. 018, OR = 1. 35, 95% CI = 1. 05 ~ 1. 74), where-as, the other results were of no statistical significance. Conclusion The results indicate that 17 SNPs may not be associated with AA in Han Chinese population. Further study should be performed in a larger Han Chinese sam-ples.

Rapid Detection of Mutations of Neisseria gonorrhoeae gyrA Gene Using DNA Chip / 中华皮肤科杂志

Objective To develop a new method-DNA chip to be used for rapid detection of mutations of Neisseria gonorrhoeae gyrA gene. Methods Probes were designed according to the sequence of Neisseria gonorrhoeae gyrA genes, and DNA chip was fabricated accordingly. DNA fragment which contains gyrA gene mutation was amplified using PCR technique, labeled with Cy5 fluorescence, and then hybridized with DNA chip. Results of DNA sequencing were used as the control. Results All of the 50 urogenital swab specimens were detected using DNA chip. The consistency between the DNA chip and drug sensitivity test, and between the DNA chip and DNA sequencing were 100% and 98%, respectively. Conclusions DNA chip is a rapid technique with high sensitivity and specificity for the detection of mutations of Neisseria gonorrhoeae gyrA gene. This method can be used for the detection of drug resistance in clinical practice.

Detection of Pathogens of Urogenital Infections and Their Drug-resistant Types by a DNA Chip / 中华皮肤科杂志

0.8). Conclusions This DNA chip combined with multiplex PCR is a rapid diagnostic assay with high specificity and sensitivity for the detection of Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma Urealyticum and their drug-resistance, and may be applied in the diagnosis of urogenital infections.