Objective To analyse preliminarily the role of amelanotic melanocutes (AMMC) and SCF/c-kit signal pathway in the pathogenesis of vitiligo. Methods Three antibodies such as HMB-45, NKl/beteb and c-kit were used to stain sections from scalps from vitiligo patients and the healthy controls. Results There were no HMB-45 positive cells in outer root sheath(ORS) of follicle. NKI/beteb positive cells were small and located in groups at the middle and lower of outer root sheath with their retraction of the dendrites. They only expressed premelanosomal antigens but not melanosomal antigen such as HMB-45. There were no significant difference of AMMC in quantities between vitiligo patients and normal controls (P>0. 05). The expression of c-kit receptors on AMMC in follicle of depigment-ed scalps from vitiligo patients was lower than that in normal contols (P<0. 05). Conclusion Abnormal c-kit expression in AMMC in the follicle of depigmented scalps may play an important role in the pathogenesis of vitiligo.

Objective To analyze the status of healthcare-associated infection(HAI)and characteristics of patho-gens in patients with traumatic brain inj ury and cerebrovascular diseases,and evaluate prevention and control meas-ures.Methods Clinical data of 236 patients with traumatic brain injury and cerebrovascular diseases in a hospital from 2008 to 2010 were analyzed retrospectively.Results A total of 29 patients developed 50 times of HAI,HAI rate was 12.29%,HAI case rate was 21.19%,HAI case rate of respiratory system,urinary system,oral cavity, gastrointestinal tract,skin and soft tissue,and other sites was 46.00%(n=23),30.00%(n=15),8.00%(n=4), 6.00%(n=3),4.00%(n=2),and 6.00%(n=3)respectively.A total of 69 pathogenic strains were detected,per-centage of gram-negative bacteria was 65.22%(n=45),the major were Pseudomonasaeruginosa(n=17),Klebsiel-lapneumoniae(n=12),Escherichiacoli(n=10),and Acinetobacterbaumannii(n=4);percentage of gram-positive bacteria was 30.43%(n=21),the major were Staphylococcusaureus(n=11),Staphylococcusepidermidis (n=7), Streptococcus spp.(n=2);percentage of fungi was 4.35%(n=3).Conclusion HAI in patients with traumatic brain inj ury and cerebrovascular diseases is high,the main infection site is respiratory system,the main pathogens are gram-negative bacteria.Preventive and control measures should be taken accordingly.

Objective To investigate the relationship between the single nucleotide polymorphisms (SNPs) of TLR9 gene and the occurrence of condyloma acuminatum (CA). Methods Peripheral venous blood was obtained from 63 patients with CA and 23 normal human controls with informed consent. DNA was extracted from the blood samples and subjected to the amplification of TLR9 gene by PCR followed by sequence analysis. Results There were 4 SNPs, i.e., SNP1, SNP2, SNP3 and SNP4 at positions 1174, 1635, 1269 and 1724 of the TLR9 gene, respectively. Of these SNPs, SNP1 was located in intron 1, SNP2, SNP3 and SNP4 in exon 2. The registration number is rs352139 for SNP1, rs352140 for SNP2 in NCBI database. SNP3 and SNP4 were newly discovered positions. The frequency at SNP1 position was 0.690 and 0.609 for allele A in the patients and controls, respectively, 0.309 and 0.391 for allele G, respectively (both P > 0.05). No significant difference was observed between the patients and controls in the frequency of allele A or allele G at position SNP2 (0.302 vs. 0.698, 0.369 vs. 0.630, both P > 0.05). There were 4 haplotypes at the SNP1 and SNP2 positions, including AA, AG, GA and GG, with no significant difference in the frequency between the patients and controls (all P> 0.05). Conclusions There are 4 SNPs including SNP1, SNP2, SNP3 and SNP4 in the TLR9 gene in Guangdong Han population. SNP1 and SNP2 appear unrelated to the liability to CA.

Background and Objectives: Melanocyte (MC), derived from neural crest stem cell (NCSC), are involved in the pro-duction of melanin. The mechanism by which NCSC differentiates to MC remains unclear. N6-methyladenosine (m6A) modification was applied to discuss the potential mechanism. Methods: and Results: NCSCs were isolated from hair follicles of rats, and were obtained for differentiation. Cell via-bility, tyrosinase secretion and activity, and transcription factors were combined to evaluated the MC differentiation.RT-qPCR was applied to determine mRNA levels, and western blot were used for protein expression detection. Total m6A level was measured using methylated RNA immunoprecipitation (MeRIP) assay, and RNA immunoprecipitation was used to access the protein binding relationship. In current work, NCSCs were successfully differentiated into MCs.Fat mass and obesity associated gene (FTO) was aberrant downregulated in MCs, and elevated FTO suppressed the differentiation progress of NCSCs into MCs. Furthermore, microphthalmia-associated transcription factor (Mitf), a key gene involved in MC synthesis, was enriched by FTO in a m6A modification manner and degraded by FTO. Meanwhile, the suppression functions of FTO in the differentiation of NCSCs into MCs were reversed by elevated Mitf. Conclusions In short, FTO suppressed the differentiating ability of hair follicle-derived NCSCs into MCs by m6Amodifying Mitf.

Seven target compounds coupled by rhein and furoxan were synthesized and their chemical structures were confirmed by 1H NMR, IR, and MS. All target compounds were evaluated for anti-proliferative activity against human hepatoma cells HepG2 and Bel-7402, human colon cancer cells HCT116, human osteosarcoma cells U2OS, drug-resistant cells Bel-7402/5-FU and normal hepatocytes cells LO2 in vitro by thiazolyl blue(MTT) colorimetry. The results indicated that all target compounds had more potent anti-proliferative activity than their parent compound rhein. Additionally, compound 4g had stronger proliferation inhibitory activity on HepG2, Bel-7402, U2OS and Bel-7402/5-FU,with little effect on the proliferation of normal cells, exhibiting selective inhibitory activity. Griess assay was used to measure the release of nitric oxide in vitro. Results showed that compound 4g could increase the releases NO in HepG2 cells, which may be associated with its antitumor effects. Furthermore, the antitumor activity of compound 4g was attenuated by NO scavenger (hemoglobin), which indicates that the antitumor activity of compound 4g may be partly related to the release of NO.

Objective To construct a domain knowledge graph of dementia care,so as to provide the foundation and guarantee for the next intelligent application based on the knowledge graph.Methods A top-down approach was adopted to construct a domain knowledge graph of dementia care.Firstly,the ontology concept is constructed from the top level,namely the schema layer of knowledge graph.Then,instances are filled,and knowledge extraction is carried out from the existing data sources,and the extracted entities and relationships are filled into the pattern layer ontology database to complete the data layer construction of the knowledge graph.Finally,the"entity relationship entity"triplet data was input into the Neo4j graph database for storage.Results In this study,the personalized care plan set of 1 012 dementia cases was used as the corpus to construct a domain knowledge graph of dementia care.The knowledge graph takes people with dementia as the core,and unfolds,one by one,around basic characteristics,care problems,and care plans in a standardized"entity-relationship-entity"triplet format,forming a large knowledge network,which contains a total of 1 522 specific dementia care knowledge entities and 8 kinds of inter-entity relationships.Conclusion The domain knowledge graph of dementia care constructed in this study clearly and intuitively shows the global pedigree and logical path of knowledge,which provides an efficient and intelligent basic guarantee for the browsing,retrieval and application of dementia care knowledge,so as to realize personalized and intelligent management of people with dementia,break through the bottleneck of lack of professionals,improve the health outcomes of people with dementia,promote the implementation of inclusive pension services,and promote healthy aging.

Objective to establish a combined diagnosis model of scleroderma related genes based on gene expression comprehensive database(GEO)and artificial neural network(ANN)and to evaluate its effect and to predict and analyze targeted traditional Chinese medicine.Methods two scleroderma chips GSE23741 and GSE95065 were obtained from the GEO database as the training group data set.Random forest and lasso regression algorithms were used to screen the key genes of scleroderma and construct the ANN model for the diagnosis of scleroderma.The validation data sets GSE76807,GSE32413 and GSE59785 were used to verify the model,and the area under curve(AUC)analysis was used to evaluate the clinical application value of ANN model.The relative expression of key gene mRNA was verified by RT-qPCR experiment.The CIBERSORT algorithm was used to estimate the bioinformatics association between scleroderma and the screened biomarkers.Finally,the key genes were used to screen the targeted traditional Chinese medicine.Results A total of 167 differential genes were obtained.Furthermore,the five most relevant key genes(SERPINE2,SFRP4,SUGCT,FBLN5,NRXN2)were screened by machine learning,and the artificial neural network diagnosis model was constructed.The model was used to draw the subject operating characteristic(ROC)curves diagnosed by the training group and the verification group,and the AUC value of the training group was 1.000.The AUC of verification group were 0.770,0.795 and 0.872 respectively.The result of RT-qPCR experiment is consistent with that of machine learning algorithm.Immune cell infiltration analysis showed that the relative content of memory CD4+T cells was significantly increased in scleroderma group,while the relative content of γ δ T cells in normal group was significantly increased.Key genes are associated with macrophage M1,T cells,memory activated CD4+T cells,resting mast cells,CD8+T cells and so on.According to the key genes,12 traditional Chinese medicines were screened.Most of the four qi and five flavors belong to warm,cold,flat,sweet,pungent and bitter,and most of them belong to the meridians of liver,spleen and lung.Conclusion the artificial neural network diagnosis model of key genes of scleroderma is constructed,which can be used in clinical diagnosis of scleroderma,and the potential targeted traditional Chinese medicine for the treatment of scleroderma is predicted,which provides a new perspective for exploring the pathogenesis of scleroderma.

Objective To evaluate the predictive value and to verify the clinical effect of JPKD-vancomycin for the trough concentration of vancomycin in patients with augmented renal clearance (ARC), and to provide a reference for clinical individualized drug therapy. Methods A retrospective analysis was conducted. The clinical data of 48 adult patients with ARC using vancomycin and monitoring steady-state trough concentration of vancomycin admitted to Suzhou Hospital Affiliated to Nanjing Medical University from July 2013 to July 2017 were collected. A combination of classical Vancomycin Calculator software and JPKD-vancomycin software was used. Based on the individual conditions of patients [gender, age, height, weight, serum creatinine (SCr), disease status], Vancomycin Calculator software was used to obtain the recommended regimen and its steady-state trough concentration, and then JPKD-vancomycin software was used to predict the steady-state trough concentration of initial regimen. If the regimen was adjusted during the treatment, JPKD-vancomycin software was used to predict the steady-state trough concentration of the adjusted regimen. The measured values of steady-state trough concentration were recorded. The weight deviation between predicted concentration and measured concentration (WRES) was calculated. WRES < 30% was considered as good prediction, and the predictive value of JPKD-vancomycin software was evaluated for vancomycin trough concentration. Results Forty-eight patients with ARC were enrolled, of whom 24 patients had adjusted the dosing regimen during the treatment. The initial concentration of blood samples was 48, after adjusting the dosage regimen, 24 blood samples were collected. The initial and adjusted daily dose of vancomycin was (2 000±500) mg/d and (2 500±600) mg/d, respectively, and the initial trough concentrations and adjusted trough concentrations was (8.4±7.3) mg/L and (9.1±4.3) mg/L, respectively. Only 14.6% and 25.0% of initial and adjusted trough concentrations reached the target range (10-20 mg/L) without significant difference (P > 0.05). The WRES value of adjusted trough concentrations predicted by JPKD-vancomycin software was significantly lower than that of initial regimen [10.6% (3.0%, 16.4%) vs. 14.3% (10.5%, 38.2%), P < 0.05], and the percentage of WRES < 30% also tended to increase [95.8% (23/24) vs. 70.8% (34/48), P < 0.05]. The well predictive rate of JPKD-vancomycin software for vancomycin trough concentration was 79.2% (57/72), but there were 15 patients with WRES > 30%. Conclusions JPKD-vancomycin software has good predictive value for the vancomycin trough concentration of ARC patients, especially for the trough concentration after adjusting the treatment regimen. JPKD-vancomycin can provide a reference for the design of clinical individualized application of vancomycin.

This study was aimed to identify pET21b-HPV16E2/BL21(DE3) strain and to optimize the expression of human papillomavirus type 16 (HPV16) E2 protein by orthogonal analysis. Four influence factors on two levels were selected to increase the target protein quantity. The four factors were induction time, induction temperature, inductor concentration and cell density. The quantity of HPV16 E2 protein was used as the evaluation parameter. Induced by IPTG, HPV16 E2 protein was analyzed by SDS-PAGE and Western Blot. Target protein was analyzed by GIS imaging system to quantify the protein level. SPSS13. 0 software was applied to analyze the result. Data showed that the expression strain pET211rHPV16 E2/BL21(DE3) was identified correctly. HPV16 E2 protein expressed mainly at insoluble form. The 42KD protein band was identified by SDS-PAGE and Western blot. Orthogonal test was applied on influence factor analysis and expression optimization successfully. Main influence factors were inductor concentration and induction temperature. The optimimum condition of maximum expression quantity was 37 degrees C, 7h, 1.0 mmol/L IPTG and OD600 1.0. In this experiment, orthogonal test could not only be used to analyze the influential factors and promote the target protein expression, but also be used to provide a better experiment method for molecular biological study.
DNA-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; metabolism ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Recombinant Proteins ; biosynthesis ; genetics

Background: Tigecycline, eravacycline, and omadacycline are recently developed tetracyclines. Susceptibility of microbes to these tetracyclines and their molecular mechanisms have not been well elucidated. We investigated the susceptibility of Moraxella catarrhalis to tigecycline, eravacycline, and omadacycline and its resistance mechanisms against these tetracyclines. Methods: A total of 207 non-duplicate M. catarrhalis isolates were collected from different inpatients. The minimum inhibitory concentrations (MICs) of the tetracyclines were determined by broth microdilution. Tigecycline-, eravacycline-, or omadacycline-resistant isolates were induced under In Vitro pressure. The tet genes and mutations in the 16S rRNA was detected by PCR and sequencing. Results: Eravacycline had a lower MIC50 (0.06 mg/L) than tigecycline (0.125 mg/L) or omadacycline (0.125 mg/L) against M. catarrhalis isolates. We found that 136 isolates (65.7%) had the tetB gene, and 15 (7.2%) isolates were positive for tetL; however, their presence was not correlated with high tigecycline, eravacycline, or omadacycline ( ≥ 1 mg/L) MICs.Compared with the initial MIC after 160 days of induction, the MICs of tigecycline or eravacycline against three M. catarrhalis isolates increased ≥ eight-fold, while those of omadacycline against two M. catarrhalis isolates increased 64-fold. Mutations in the 16S rRNA genes (C1036T and/or G460A) were observed in omadacycline-induced resistant isolates, and increased RR (the genes encoding 16SrRNA (four copies, RR1-RR4) copy number of 16S rRNA genes with mutations was associated with increased resistance to omadacycline. Conclusions Tigecycline, eravacycline, and omadacycline exhibited robust antimicrobial effects against M. catarrhalis. Mutations in the 16S rRNA genes contributed to omadacycline resistance in M. catarrhalis.