Objective To construct yeast cDNA expression library of human dermal papillae cells (DPCs) in primary culture.Methods Human dermal papilla cells (DPCs) were isolated by two-step digestion method and cultured in DMEM medium.Total RNA was extracted from primary DPCs that exhibited an aggregative behavior in culture,then,cDNA was synthesized and amplified by using CloneMinerTM cDNA Library Construction kit to construct primary cDNA library and yeast cDNA expression libary.Results The average titer and total clones were 7.0×106 colony forming units(cfu)/ml and 1.4×107 cfu respectively in the primary library,5.5×106 cfu/ml and 1.1×107 cfu respectively in the yeast expression library.The average insert size Was 1.2 kb and the recombination rate was above 95%.Conclusions The yeast cDNA expression library of DPCs in primary culture has been successfully constructed.which will lay a foundation for screening proteins interacting with HSPC016 gene in DPCs with yeast two-hybrid system.

To train medical students' clinical thinking ability is an important aspect of clinical probation practice and also the common goal of dermartology probation training. This article analyzes the problems around the cultivation of clinical thinking ability during probation practice in our department,and proposes the corresponded measures to improve it.

Objective To detect the expression and function of cysteinyl leukotriene receptors (CysLTRs)in keratinocytes.Methods Human keratinocytes were isolated from the tissue of foreskin by digestion with dispase Ⅱ and trypsin,and subjected to primary culture.By using confocal laser scanning microscopy and reverse transcriptase PCR,the localization and expression of CysLTRs were studied in kemtinocytes.respectively.Some primarily cultured keratinocytes were pretreated with leukotriene D4 (30 nmoi/L),MK571(300 nmol/L),and BAYu9773 for 5 minutes followed by the detection of intmcellular calcium level using the Ca2+ indicator dye Fura-2/AM as well as cell proliferation bv MTT assay.Results The expressions of CysLTR1 and CysLTR2 were observed in cultured keratinocytes,and they were mainly located on cell membrane,partly in cytoplasm and nuclei.Compared with non.stimulated cells,a significant increase Was noted in the expression of CysLTRs,especially in the nuclei of keratinocytes stimulated by LTD4(P＜0.05),together with an elevation in intracellular calcium level(42.27±3.00 mmol/L,P＜0.01)and acceleration in cell proliferation (P＜0.01).However,both MK571 and BAYu9773 could completely block the effect of LTD4 on intmcellular calcium level and cell proliferation.and there was no significant difference in the blocking effect between MK571 and BAYu9773.Conclusions Functional CysLTRs are expressed in human keratinocytes.and they carl increase the intracellular calcium level in,and cell proliferation of,keratinocytes.

Objective To compare the efficacy and safety of excimer laser 308 nm phototherapy alone and the combination of excimer laser 308 nm and topical application of vitamine D3 alanogue tacalcitol in the treatment of vitiligo. Methods Seventy-eight patients with vitiligo were enrolled in the single-blind clinical trial, treated with excimer laser 308 nm. The lesions were devided into two groups: patients in the experimental group were instructed to use tacalcitol ointment and the control group were applied with placebo ointment. The lesions were evaluated once per month and photos taken for analyses of clinical effects. Results The results in different locations were compared, the effective rates of the experimental group in cephalofacial site, trunk and limbs were 93.51%, 84.16 % and 42.35 %, respectively. The effective rates of control group in opposite and adjacent sites were 90.9 %, 77.45 % and 34.15 %, respectively (P ＜ 0.05). The comparison of results in different types of lesions indicated that the effective rate of the experimental group in vitiligo vulgaris and segmental vitiligo were 73.81% and 84.00 %, respectively. The effective rate of control group in vulgaris and segmental vitiligo were 86.8 %, 77.45 % and 34.15 %, respectively (P ＜0.05 ). The comparison of results in radiation times and doses of phototherapy showed that the radiation time and dose on the time of initial pigment regeneration were (16. 15 ± 3.22)times and (4.40 ± 5.03)J/cm2 in the experimental group, while ( 18.56 ± 3.50) times and ( 6.60 ± 1.01 ) J/cm2 ( P ＜ 0.05 ) in the control group, the time and dose on the time of apparent effect were ( 20. 36 ± 1.50 ) times and ( 7.50 ± 3.54 ) J/cm2 in the experimental group, and (21.68 ± 2.40) times and( 8.80 ± 9.24)J/cm2 (P ＜ 0.05 ) in the control group. Conclusion Application of tacalcitol ointment in combination with twice-weekly 308 nm excimer laser light phototherapy is an effective alternative treatment for patients with generalized vitiligo.

Objective To study the effects of recombinant chimeric toxin Dsg3EC_(1-2)PE40 on T and B cells from PV patients. Methods The recombinant protein Dsg3EC_(1-2)PE40 was expressed on BL21TrxB (DE3) cells, then identified and purified. ELISPOT assay was used to detect and quantitate autoantibody-producing B cells in different concentrations of recombinant chimeric toxin, and MTT assay and ~3H-TdR assay to observe the metabolism and proliferation of T cells from PV patients in vitro. Results The purity of expressed protein Dsg3EC_(1-2)PE40 was up to 80%. The number of anti-Dsg3 antibody-producing B cells in PBMC from PV patients decreased by 40% with treatment of Dsg3EC_(1-2)PE40, which was significantly lower (P

Objective To evaluate the efficacy and safety of olopatadine hydrochloride for the treatment of chronic idiopathic urticaria (CIU). Methods A multicentre, double-blind, randomized, parallel-group, controlled clinical trial was conducted. A total of 144 patients with CIU from 3 research centers were enrolled into this study, and randomly and equally divided into a test group and a control group. The test group administrated olopatadine hydrochloride 5 mg twice a day for 28 consecutive days, while the control group administrated levocetirizine hydrochloride 5 mg in the forenoon and a placebo tablet of olopatadine hydrochloride 5 mg in the afternoon for 28 consecutive days. The symptom score reducing index(SSRI)served as the primary outcome, and global assessment score for efficacy and total response rates as the secondary outcome. Results Totally, 137 patients completed the trial, including 70 in the test group and 67 in the control group. As intention-to-treat analysis showed, there were no significant differences in the total response rate between the test group and control group on day 7 (64.29% (45/70)vs. 56.72%(38/67), P > 0.05), 14(82.86%(58/70)vs. 74.63%(50/67), P > 0.05), or 28(87.14%(61/70)vs. 77.61%(52/67), P >0.05)after start of treatment. The SSRI was significantly higher in the test group than in the control group after 4 weeks of treatment(82.67% ± 22.70% vs. 70.51% ± 32.07%, P < 0.05). In addition, no significant difference was observed in the incidence of adverse reactions between the test group and control group(33.80%(24/71)vs. 27.94%(19/68), P > 0.05), and adverse reactions mainly included lethargy, dry mouth, fatigue, etc. Conclusion Olopatadine hydrochloride is effective and safe for the treatment of CIU.

[Objective] To evaluate the efficacy of autologous whole blood injections in patients with chronic spontaneous urticaria and positive autologous serum skin test (ASST).[[Methods]] After assessment of clinical history,patients with chronic spontaneous urticaria underwent skin prick test (SPT) and ASST.Then,100 patients with positive ASST but negative SPT for common allergens were randomly classified into treatment group (n =60) and control group (n =40).Oral loratadine was given to all the patients with a gradual tapering to the least maintenance dose.Patients in the treatment group were also injected with autologous whole blood once a week for 12 times.Patients were evaluated by urticaria activity score (UAS) and dermatology life quality index (DLQI) at the baseline,the end of the 3rd and 6th month after the initial treatment.The total amount of antihistamines required for the control of urticaria every month was calculated.The UAS,DLQI,accumulative amount of administrated antihistamines,and the diameter of wheal/flush induced by autologous serum were compared by t test before and after the treatment,and the efficacy was compared by rank sum test between the two groups.[Results] No significant difference was observed between the control and treatment group in UAS at the baseline (5.73 ± 0.51 vs.5.32 ± 0.79,P＞ 0.05).The UAS reached 1.57 ± 1.42 and 0.69± 0.92 with a decrease rate of 69％ and 81％ in the treatment group,and 3.65 ± 1.53 and 2.65 ± 1.61 with a decrease rate of 35％ and 53％ in the control group,respectively at the end of the 3rd and 6th month,and statistical difference was observed for the decrease in both groups at the two time points (all P ＜ 0.05).The total amount of antihistamines required for the control of urticaria per month averaged 8.63 pills and 3.83 pills respectively in the treatment group after 3 and 6 months of treatment,significantly less than that in the control group (16.85 and 15.27 pills,respectively).[Conclusion]s The combination of oral antihistamine and autologous whole blood injections can not only reduce disease activity and improve patients' quality of life,but also decrease the total amount of antihistamines required for the control of urticaria.

Objective To assess the association between rs7597593 polymorphism of ZNF804A gene and schizophrenia,and to assess the relationship between rs7597593 polymorphism and working memory.Methods Schizophrenia patients and healthy controls were diagnosed in accordance with Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-Ⅳ) ; 767 schizophrenia patients and 690 healthy controls were involved.Restriction fragment length polymorphism (RFLP) was carried out to genotype rs7597593 polymorphism.The cognitive function of working memory was assessed by the N-back task.Statistical analyses were carried out with SPSS19.0 software.Results The study found no significantly different genotype frequencies (x2=1.519,P=0.468) and allele frequencies(x2=1.263,P=0.261) of rs7597593 polymorphism between schizophrenia patients and healthy controls,however in the subgroup of higher IQ (IQ ≥ 110),there were significant different distributions of both genotype and allele (x2 =9.411 and 6.529; P=0.009 and 0.011 respectively).It was also found in this subgroup that risk T allele was associated with more error at 1-back task (F=6.854,P=0.009).Conclusion These results indicated that rs7597593 polymorphism was associated with individuals having spared cognitive function; carriers of T allele had worse cognitive function,which maybe a pathway that it contributes to schizophrenia.

Pseudoxanthoma elasticum(PXE) is a rare, genetic, metabolic disease characterized by ectopic calcification of connective tissue that primarily affects the skin, retina and cardiovascular system, which characteristic histopathology is calcification and fragmentation of elastic fibers in dermis.PXE is mainly caused by ABCC6 gene mutation, which is one of the important regulators of the serum inorganic pyrophosphate (PPi) homoeostasis, a main inhibitor of ectopic calcification and the deficiency of PPi can lead to ectopic calcification. The clinical features are highly heterogeneous.Typical skin lesions of PXE are yellowish flat papules and plaques, and the symptoms of skin relaxation and shrinkage can be manifested in the later stage.Retina, cardiovascular and other complications seriously affect the health and quality of life of patients. The current therapy of PXE include symptom improvement, systemic anti-ectopic calcification medicine, gene therapy and so on.We review the pathogenesis, clinical manifestations, diagnosis and treatment of PXE.

Objective:To analyze the differentially expressed genes in PBMCs of patients with systemic lupus erythematosus (SLE) by bioinformatics methods screening and analyzing the key genes related to ferroptosis, and explore the possible mechanism of ferroptosis involved in the pathogenesis of SLE at the transcription level.Methods:The data sets and samples of healthy people (HC) and SLE patients who met the screening criteria were retrieved from the Gene Expression Omnibus (GEO), a sub-database of the National Center for Biotechnology Information (NCBI). The differentially expressed genes, GO enrichment analysis and KEGG pathway enrichment analysis were analyzed by GEO2R, R language and related software packages. The protein interaction network (PPI) of differential genes was analyzed by STRING, Cytoscape and other tools to explore the key genes and pathways. In addition, real-time quantitative reverse transcription PCR (RT-qPCR) was used to verify the expression of key genes. Mann-Whitney U test was used to compare the expression of key genes in PBMCs between the two groups. Spearman rank correlation analysis was used to explore the relationship between SLE disease activity and the level of key genes. Results:Six data sets were included in this study. A total of 166 genes related to ferroptosis were differentially expressed between SLE and HC groups. The differential genes were specifically expressed in alveolar macrophages, neutrophils, CD49 + cells and CD31 + cells. GO enrichment analysis and KEGG pathway enrichment analysis showed that the differentially expressed genes were mainly involved in multiple signaling pathways closely related to SLE, such as oxidative stress response, infection and TNF signaling pathway. Hub genes screened by different algorithms all suggested RELA as a key gene, and RT-qPCR confirmed that compared with the RELA gene expression level in the HC group [0.75(0.37,1.13)], the expression level in SLE group [2.02 (1.19,4.06)] was increased, the difference was statistically significant ( Z=-3.08, P=0.002), and was positively correlated with the corresponding SLEDAI score of SLE samples ( r=0.52, P=0.019). Conclusion:The ferroptosis of many immune cells, including alveolar macrophages and CD49 + NK cells, is involved in the pathogenesis of SLE. RELA may be involved in the ferroptosis of PBMCs in SLE through the NF-κB pathway.