Objective To analyze the mutations in keratin 1 (KRT1), KRT9 and KRT10 genes in a Chinese family with epidermolytic palmoplantar keratoderma (EPPK). Methods Clinical data were collected from a family with EPPK. Genomic DNA was extracted from the peripheral blood of 12 family members, including 6 patients and 6 unaffected members, as well as from 50 unrelated normal human controls. PCR was performed to amplify all the exons and flanking sequences of KRT1, KRT9 and KRT10 genes followed by DNA sequencing.Results A missense mutation C.1436T > C was found in the highly conserved helix termination motif of KRT1 gene of all the patients, resulting in a substitution of isoleucine by threonine at position 479 of the KRT1 protein. No mutation was found in the unaffected members or unrelated controls. Conclusions The missense mutation C.1436T > C in K.RT1 gene is likely to be the main cause of the phenotype of EPPK in this family.This is the first report of a pedigree with KRT1 gene mutation-induced EPPK in China.

Objective To analyze the influence of mode of delivery on post-neonatal gut microbiota using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technology.Methods From April to August in 2013,thirty healthy urban full-term neonates in Nanjing City were enrolled in the study,including fifteen exclusive breastfed ones (seven born of caesarean section and eight born vaginally) and fifteen mixed feeding ones (eight born of cesarean section and seven born vaginally).Stool specimens were collected on the 28th day after birth and bacterial genomic DNA was extracted and examined by PCR on 16S rDNA V3 variable region.Bacterial community profiles were obtained by DGGE.Diversity and similarity differences of the gut microbial community structures were analyzed.Two independent sample t test or Chi-square tests were used for stastistical analysis.Results (1)Diversity analysis showed that among exclusive breastfeeding infants,the Strip number and Shannon-Weaver Diversity Index of gut microbiota in infants born abdominally were significantly lower than those born vaginally [9.71 ±4.27 vs 15.12±4.19,2.13±0.39 vs 2.61±0.32,both P＜0.05],but the Simpson index of gut microbiota was significantly higher [0.13 ± 0.04 vs 0.08± 0.03,P＜0.05],and no significant difference was shown in Pielou Index (P＞0.05).In the mixed feeding group,the Strip number and Shannon-Weaver Diversity Index of gut microbiota in infants born abdominally were significantly lower than those born vaginally [10.88±3.23 vs 16.29±5.38,2.26±0.37 vs 2.66±0.31,both P＜0.05],the Simpson index was higher,but together with the Pielou Index,neither showed significant difference (both P＞0.05).(2) Similarity analysis found that gut microbiota from neonates born of same mode of delivery mostly gathered together and had much more similar structures.Conclusions In the post-neonatal period,the species and numbers of gut microbiota in infants born abdominally are all behind of those born vaginally.The predominant microbiota in babies born of cesarean section are more prominent,and gut microbiota in vaginal delivered babies are more uniformly distributed.

@@

Objective To test the possibility that the thoracic defomity is a risk factor of the diopathic pneumothorax in youth by making a statistical analysis of the thoracic sagittal diameter and transverse diameter ratio in patients and normal people. To study the correlation between this ratio and age of the patients. Methods 107 cases of idiopathic pneumothorax were taken as subjects and other 105 cases in' the same period with normal pneumothorax were used as the control group. T-test,correlation study and scottered spot maps were made based on the ratio of vertical and horizontal diameters,the mean and the age. Results The mean thorax aspect ration( MTAR) of the patients with spontaneous pneumothorax was 0.38, and MTAR in normal people was 0.44, indicating a statistical significance ( P < 0.01) of the difference of MTAR between the two groups. The age of the patients' first onset age and the ratio of vertical and horizontal diameter was positively correlated. Conclusion Thoracic deformity of idiopathic pneumothorax had a higher incidence in young people,and there was a positive correlation between of the age of their first onset and the ratio of vertical and horizontal tracks.

Objective To explore the correlation between circadian clock gene clock circadian regulator (CLOCK) and attention-deficit/hyperactivity disorder (ADHD) and the role of CLOCK and sleep problems on inhibition in male children with ADHD. Methods Two single nucleotide polymorphisms (SNPs) of CLOCK were genotyped in 854 male ADHD children and 320 male controls. Sleep problems were assessed using parent symptom questionnaire. In ADHD cases, the main effects and interaction of CLOCK SNPs and sleep problems on inhibition assessed by using Stroop Color and Word Test, were analyzed using the analysis of covariance (ANCOVA). Results No significant differences of allele and genotype frequencies were found for rs6832769 and rs11932595 in all case-control groups (P>0.05). In ADHD cas?es, the main effects of rs6832769 and rs11932595 genotypes and sleep problems on inhibition were not significant (P>0.05). However, the interaction of rs6832769 genotype and sleep problems was significant (F=6.71, P=0.01). When ac?companied with sleep problems, ADHD cases carrying the AA&AG genotype showed the longest time of word interfer? ence (F=6.63, P=0.01). Conclusions Inhibition of male ADHD children can be modulated by the interaction of CLOCK rs6832769 and sleep problems.

Objective To explore the gender difference in clinical features and outcomes of elderly patients with coronary true bifurcation lesions after percutaneous coronary intervention (PCI).Methods A total of 169 consecutive patients were included in the study, who were diagnosed coronary true bifurcation lesions by coronary angiography (CAG)and received PCI in our hospital from December 2013 to December 2015.All patients were divided into 2 groups according to their gender (71 femals and 98 males).Angiographic characteristics of the coronary lesions and clinical data were analyzed in both groups.Clinical outcomes during follow up were recorded and studied.Results The levels of total cholesterol (TC), triglyceride (TG),high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) in the female group were significantly higher than in the male group (all P<0.05).The levels of BUN,Cr,UA,CyC and TBIL in the female group were significantly lower than in the male group (all P<0.05).Comorbidity rates of hypertension and diabetes were higher in female patients without significant differences (P>0.05).The median follow-up time was 17 months and the incidence rate of overall adverse events in women was higher than that of men (25.4% vs.11.2%, P<0.05).Conclusions The clinical outcomes of elderly patients with coronary true bifurcation lesions after PCI had gender differences.The rates of adverse events in female patients was found significantly higher than male patients.

Objective To evaluate the efficacy of benvitimod on allergic contact dermatitis (ACD) in a mouse model of allergic contact dermatitis.Methods Acute and chronic ACD models were established respectively in 42 BALB/c mice through 2,4-dinitrofluorobenzene (DNFB) sensitization and challenge.Then,the BALB/c mice were equally divided into 7 groups with 6 mice in every group:normal control group receiving no treatment,five treatment groups topically treated with 0.1％ dexamethasone solution,0.1％ tacrolimus (FK506) solution,0.5％ benvitimod solution,1.0％ benvitimod solution and 2.0 ％ benvitimod solution respectively,and dehydrated alcohol group treated with dehydrated alcohol only.All the drug solutions were topically applied at 24 and 36 hours after the last challenge in the murine models of acute ACD which were sacrificed at 48 hours,and twice daily from day 7 to 21 after the initial sensitization in the murine models of chronic ACD which were sacrificed on day 21 after the first topical treatment.Ear tissues were obtained from these mice and subjected to measurement of ear thickness and weight,as well as pathological examination and evaluation of inflammatory infiltrate by hematoxylin-eosin (HE) staining.The safety of these drugs was also estimated at the end of treatment.Results In the murine models of acute ACD,benvitimod showed no obvious therapeutic effect at 24 hours,with no significant differences in bilateral difference in ear thickness or weight between the three benvitimod groups and dehydrated alcohol group (all P ＞ 0.05).Meanwhile,in the murine models of chronic ACD,benvitimod markedly decreased the swelling degree of ears,with significant differences between the three benvitimod groups (0.5％,1.0％ and 2.0％) and dehydrated alcohol group in bilateral difference in ear thickness ((71.50 ± 3.15) × 10-3 mm,(75.50 ± 3.02) × 10-3 mm and (69.50 ± 2.59) × 10-3 mm vs.(91.83 ± 2.04) × 10-3 mm,all P＜ 0.01) and weight ((2.33 ± 0.45) mg,(2.30 ± 0.57) mg and (2.38 ± 0.27) mg vs.(3.73 ± 0.33) mg,all P ＜ 0.01) after 3 weeks of treatment.The inflammatory infiltration in ear tissue was significantly attenuated in murine models of both acute and chronic ACD by the three concentrations of benvitimod compared with dehydrated alcohol (all P ＜ 0.01).Conclusions Topical benvitimod can inhibit chronic ACD in mice induced by 2,4-DNFB,but exhibits no obvious effect on acute ACD.No apparent local adverse effects were observed during the treatment with benvitimod in these mice.

Abstract: Objective To construct a shuttle vector pHT315-AaCPR100A with two spore-producing-dependent promoters and the target gene AaCPR100A in Escherichia coli-Bacillus thuringiensis. Methods The forward promoter of Cry3A, named Pro-1 (+), was amplified by PCR using pSVP27A plasmid as the template, and the target gene AaCPR100A was amplified using Aedes aegypti RNA reverse conversion cDNA as the template. The plasmid pHT315 was linearized by digestion with Hind Ⅲ and Sal Ⅰ. The forward promoter and the target gene were inserted into the linearized vector pHT315 successively by in-fusion cloning according to the transcription direction. The synthesized plasmid containing the Cry3A reverse promoter sequence was used as the template, and the Pro-1 (-) reverse promoter was amplified by PCR. The intermediate vector containing the forward promoter and the target gene was linearized by EcoR I restriction enzyme, and the reverse promoter was inserted downstream of the target gene by in-fusion cloning in the direction of transcription. Results By agarose gel electrophoresis, the forward promoter, target gene AaCPR100A and reverse promoter bands were clear and of good quality, which could be used for in-fusion cloning experiments. The two spore-producing-dependent promoters and target gene fragments were connected by In-fusion cloning. The recombinant vector pHT315-AaCPR100A was verified by PCR. The forward promoter, target gene fragment and reverse promoter were successfully amplified in the recombinant vector. Nucleotide sequencing verified that the sequencing results of the bidirectional promoter sequence and the target gene sequence were basically consistent with the sequence alignment results, which met the requirements of the construction of vector elements and proved that the recombinant vector was successfully constructed. Conclusions Based on the above results, this study proves that the recombinant shuttle vector with two spore-producing-dependent promoters can be successfully constructed by in-fusion cloning technology, laying the foundation for the construction of engineered Bacillus thuringiensis expressing dsRNA of AaCPR100A.

Objective:To identify the hub differentially expressed genes(DEGs)of glomerular pathological changes and potential pathways in molecular process of type 2 diabetic nephropathy(DN)based on bioinformatics technology.Methods:The differentially expressed genes of Gene Expression Omnibus(GEO)dataset GSE96804 in DN and normal kidney tissues were analyzed by R 3.6.2 software. DEGs were further assessed by Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway analysis. Subsequently, the hub genes and their associated pathways were analyzed using String 11.0 and Cytoscape 3.7.2 software.Results:A total of 168 DEGs were obtained in the dataset. Among them, seven hub genes were identified, including ALB, FN1, EGF, PTGS2, PLG, KDR, and LOX. Three hub genes, ALB, EGF, PLG, exerted a direct action on glomerulus. GO enrichment analysis of DEGs was mainly manifested in extracellular matrix organization, extracellular structure organization, platelet degranulation and other biological processes, extracellular matrix, secretory granule lumen, platelet alpha granule and other cell components, chaperone binding, copper ion binding, antioxidant activity, and other molecular functions. DEGs mainly regulated metabolic process, which was related to fatty acid degradation signal pathway, exogenous substance metabolism related to CYP enzyme and drug metabolism signal pathway.Conclusion:A bioinformatics analysis of DN from the perspective of glomerulopathy is helpful to understand the potential molecular mechanism of DN and provide reference for further validation.

The study aims to investigate the associations of SUMO4 polymorphisms with tacrolimus concentrations in Chinese renal transplant recipients. Blood samples and clinical data were collected from 132 renal transplant recipients with tacrolimus treatment. CYP3A5*3 genotypes were detected by PCR-RFLP, and SUMO4 (rs237024, rs237025) genotypes were detected by Sequenom® MassARRAY system. SUMO4 rs237024 and rs237025 genotypes were in complete linkage disequilibrium (D' = 1). The dose-adjusted concentration of tacrolimus in SUMO4 rs237024A-rs237025A (GA-GA +AA-AA) carriers was considerably higher than that in GG-GG carriers (P < 0.05). After stratification by CYP3A5*3 genotypes, SUMO4 rs237024A-rs237025A carriers (GA-GA+AA-AA) had a higher dose-adjusted tacrolimus concentration than that in GG carriers in CYP3A5 expresser (P < 0.05). The results illustrated that SUMO4 rs237024 and rs237025 polymorphisms were associated with tacrolimus concentrations, and the test of these genotypes may be useful for individualized medicine of tacrolimus.