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BACKGROUND@#Systemic lupus erythematosus (SLE) is a complex autoimmune disease, and the mechanism of SLE is yet to be fully elucidated. The aim of this study was to explore the role of two-pore segment channel 2 (TPCN2) in SLE pathogenesis.@*METHODS@#Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of TPCN2 in SLE. We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell. Knockdown of TPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting. Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation, apoptosis, and cell cycle of TPCN2-deficient cells. In addition, gene expression profile of TPCN2-deficient cells was analyzed by RNA sequencing (RNA-seq).@*RESULTS@#TPCN2 knockdown with short hairpin RNA (shRNA)-mediated lentiviruses inhibited cell proliferation, and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells. We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells, and screened the differential genes, which were enriched for the G2/M checkpoint, complement, and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways, as well as changes in levels of forkhead box O, phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin, and T cell receptor pathways; moreover, TPCN2 significantly influenced cellular processes and biological regulation.@*CONCLUSION@#TPCN2 might be a potential protective factor against SLE.
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Humanos , Apoptosis/genética , División Celular , Células Jurkat , Lupus Eritematoso Sistémico/genética , ARN Interferente Pequeño/genéticaRESUMEN
Objective To measure the thickness and echo density of the skin at multiple sites of healthy adults by using high-frequency ultrasound.Methods A total of 50 healthy volunteers were enrolled from Department of Dermatology and Venereology,China-Japan Friendship Hospital between June and December in 2018,including 33 females and 17 males aged 22-69 years.The thickness and echo density of the epidermis,dermis and epidermis-dermis layer were detected by using high-frequency ultrasound at 12 sites,including the forehead,cheek,chest,abdomen,and the inner and outer sides of the upper arm,forearm,thigh and leg.The means of two groups were compared by using t test,and means of several groups were compared by using one-way analysis of variation.Results High-frequency skin ultrasound images differed among different anatomical sites.There were significant differences in the thickness and echo density of the epidermis,dermis and epidermis-dermis layer among the 12 sites (P ＜ 0.05).The epidermis was thickest at the inner side of the thigh (160.68 μm ± 25.71 μm),the dermis was thickest at the cheek(1 828.78 μm ± 399.10 μm),and the epidermis-dermis layer was thickest at the cheek (1 943.48 μm ± 402.4 μm).The echo density of the epidermis,dermis and epidermis-dermis layer was highest at the inner side of the leg (152.27 ± 21.56),forearm (52.71 ± 15.57) and forearm (62.56 ± 15.76) respectively.The thickness of the epidermis,dermis and epidermis-dermis layer at the forehead,cheek and the inner side of the forearm was significantly higher in male volunteers than in female volunteers (P ≤ 0.05 or ＜ 0.01),while the echo density of the dermis and epidermis-dermis layer at the forehead,cheek and the outer side of the leg was significantly lower in male volunteers than in female volunteers (P ＜ 0.05 or 0.01).Conclusion Differences exist in skin thickness and echo density among different anato mical sites and between male and female healthy adults.
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Objective To evaluate the diagnostic value of dermoscopy and reflectance confocal microscopy (RCM) alone or in combination for melanocytic nevus.Methods A total of 37 patients with clinically diagnosed melanocytic nevus were collected.Skin lesions were firstly examined by dermoscopy and RCM,then were resected to be subjected to histopathological examination for final diagnosis.The imaging features of melanocytic nevus were summarized.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of different skin imaging techniques were calculated,and the consistency was analyzed between skin imaging techniques and histopathological examination.Results Based on the dermoscopic and RCM findings,2 kinds of nevus cells with different morphological features were observed in the dermis of intradermal nevus.One kind of nevus cells was characterized by a nonfusional,highly-refractive round structure in the papillary dermis under RCM,and by a brown or light brown homogenous pattern under dermoscopy,which was observed in 5 skin lesions.The other kind of nevus cells appeared as irregular,highly-refractive cell clumps in the papillary dermis under RCM,and by a cobblestone or globular pattern under dermoscopy,which was observed in 31 skin lesions.For the diagnosis of melanocytic nevus,the sensitivity,specificity,accuracy,positive predictive value and negative predictive value of RCM combined with dermoscopy were 91.7％,87.5％,90.9％,97.1％ and 70％ respectively,those of RCM were 86.1％,75％,84％,93.9％ and 54.5％ respectively,and those of dermoscopy were 77.8％,87.5％,75％,96.3％ and 41.2％ respectively.All the diagnostic indices of RCM combined with dermoscopy were higher than those of RCM or dermoscopy alone,except that the specificity was equal to that of dermoscopy alone.RCM showed higher sensitivity,accuracy and negative predictive value,but lower specificity and positive predictive value compared with dermoscopy.There were no significant differences in the diagnostic yield in melanocytic nevus between RCM combined with dermoscopy or RCM alone and histopathological examination (x2 =0.25,0.57,P =0.63,0.45,Kappa value =0.72,0.53,respectively).However,a significant difference in the diagnostic yield in melanocytic nevus was observed between dermoscopy and histopathological examination (x2 =5.81,P =0.012).Conclusion RCM combined with dermoscopy shows higher diagnostic accuracy for melanocytic nevus compared with RCM or dermoscopy alone.
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Objective To investigate in vitro combined effect of ceftriaxone and azithromycin against clinical isolates of Neisseria gonorrhoeae(NG). Methods A total of 25 NG clinical isolates were collected from the STD clinic of Dalian Dermatosis Hospital in 2012. Epsilometer test(Etest)method was used to determine the minimum inhibitory concentrations(MICs)of ceftriaxone and azithromycin against NG isolates. Fractional inhibitory concentration index (FICI) was calculated to evaluate the in vitro combined effect of ceftriaxone and azithromycin against NG isolates. Results The mean MICs of ceftriaxone and azithromycin were 0.032 mg/L (range, 0.008- 0.064 mg/L) and 0.834 mg/L (range, 0.064-4.000 mg/L), respectively. The FICI ranged from 0.724 to 2.696, and ceftriaxone and azithromycin showed an additive effect against the above NG isolates. Conclusion Ceftriaxone and azithromycin show an additive effect against NG in vitro, but further studies with large sample size are needed to confirm their effects.
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Objective To compare the effects of medroxyprogesterone acetate (MPA) and compound norethisterone enanthate (CNE) on the susceptibility of BABL/c mice to lower reproductive tract infection with chlamydia trachomatis (Ct). Methods A total of 60 BALB/c mice were randomly and equally divided into 6 groups:MPA-pretreated control group and CNE-pretreated control group inoculated with MyCoy cell suspensions in the vagina on the 5th day after single treatment with MPA and CNE respectively, blank control group receiving no treatment, MPA-pretreated infected group and CNE-pretreated infected group inoculated with 1 × 107 inclusion-forming units(IFU)of Ct serovar E in the vagina on the 5th day after single treatment with MPA and CNE respectively, control infected group inoculated with the same quantity of IFU of Ct serovar E in the vagina but receiving no pretreatment. On day 4, 7 and 14 after inoculation, vaginal irrigation fluid was obtained from all the mice for cell culture of Ct. Three mice were randomly selected from each of these groups at the above three time points and sacrificed, and vaginal and uterine tissue specimens were obtained for hematoxylin-eosin(HE)staining and microscopic examination. Chi-square test and Fisher's exact test were conducted to compare infection rate among different groups. Results No growth of Ct was observed in the three control groups at the above time points. The culture-positive rate of Ct was 1/10 on day 4 but 0 on day 7 and 14 in both the CNE-pretreated infected group and control infected group, 7/10 on day 4, 2/7 on day 7 but 0 on day 14 in the MPA-pretreated infected group. Fisher's exact test revealed that the culture-positive rate of Ct was significantly higher in the MPA-pretreated infected group than in the control infected group and CNE-pretreated infected group on day 4 (both P =0.03), but similar among the three infected groups on day 7 (P = 0.23). Both the MPA-pretreated control group and infected group showed an increase in endovaginal mucus, thinning of vaginal stratified squamous epithelium, mucification of vaginal epithelium, presence of secretions in vaginal lumen and submucosal infiltration of a few inflammatory cells on day 4, 7 and 14, as well as appearance of pathological changes (including the presence of large quantities of purulent secretions in lumen, mild tissue edema and submucosal infiltration of a few inflammatory cells) in the vagina on day 4. Vaginal tissues were normal in both the CNE-pretreated infected group and control group at the above three time points, but mild tissue edema, lumen expansion, secretion retention and infiltration of scattered inflammatory cells were observed in the uterus on day 4 after inoculation. Conclusions MPA can arrest the estrous cycle of mice at diestrus with the mucification of vaginal epithelium, which may increase the susceptibility to Ct vaginal infection in mice. In contrast, CNE has no obvious effect on the estrous cycle and susceptibility to Ct vaginal infection despite of the appearance of pathological changes in the uterus.
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Objective To test the ceftriaxone susceptibility of Neisseria gonorrhoeae (NG) isolates from Nanjing city,and to assess their genotypes by using the NG multi-antigen sequence typing (NG-MAST) method.Methods A total of 204 NG strains isolated in 2007 and 81 in 2012 from Nanjing city were included in this study.The minimum inhibitory concentration (MIC) of ceftriaxone was determined for these strains using an agar dilution method.DNA was extracted by the Qiagen commercial kit from these strains followed by NG-MAST.Results All the isolates were susceptible to ceftriaxone (MIC,≤ 0.25 μg/ml).The MIC of ceftriaxone was ≥ 0.06 μg/ml for 63.2％ of all the NG strains,70.6％ of those isolated in 2007 and 44.4％ of those in 2012,and ≥ 0.125 μg/ml for 31.6 ％ of all the NG strains,39.7％ of those isolated in 2007,11.1％ of those in 2012.Totally,166 genotypes were identified among the 285 isolates,of which,73 had been reported,and 93 were previously unreported.The most prevalent genotype was ST568 (n =13) in NG strains isolated in 2007,followed by ST270 (n =9),ST421 (n =7),ST2288 (n =5),ST1731 (n =4),ST1766 (n =4),ST1866 (n =4),ST1870 (n =4),while ST2318 (n =5),ST1053 (n =4),ST5990 (n =4),ST8726 (n =4) were the common genotypes in 2012.Those isolates with identical or similar genotypes tended to display similar MICs for ceftriaxone.Conclusions The prevalent genotypes of NG are markedly different between 2007 and 2012 in Nanjing region,and there is a strong association between the genotypes and ceftriaxone susceptibility of NG.NG-MAST results may serve as a genetic marker in the surveillance of antibiotic susceptibility in NG.
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Objective To compare the pathogenicity between Ureaplasma urealyticum serotype 1 (Up1)and 8 (Uu8) in the genital tract of BALB/c mice.Methods A total of 48 BALB/c mice were randomly and equally divided into four groups:blank control group receiving no treatment,estradiol group pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with sterial liquid culture media,Up1 and Uu8 groups pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with suspensions of Up1 and Uu8 respectively.Three mice were randomly selected from each group to be sacrificed after the collection of vaginal lavage fluid on day 3,7,14 and 21 after the inoculation.Vaginal and uterine tissue specimens were obtained from these sacrificed mice and underwent hematoxylin and eosin (HE) staining.Vaginal lavage fluid samples were subjected to culture of Uu and measurement of tumor necrosis factor-α (TNF-α).Results No evidences were observed for Uu growth in either the blank control group or estradiol group at any of the time points after the inoculation,with the average level of TNF-α in vaginal lavage fluid being (4.17 ± 0.85) pg/ml at these time points in both groups.Uu grew in all the vaginal lavage fluid samples from the Up1 and Uu8 groups at the four time points,with the color change unit (CCU) value decreasing with time.The level of TNF-α in vaginal lavage fluid peaked on day 14 after the inoculation in the Up 1 ((14.93 ± 1.11) pg/ml) and Uu8 ((27.04 ± 24.26) pg/ml) groups.Both Up1 and Uu8 infection caused acute and chronic inflammatory responses in the mice,which were mainly located in the uterus,and Up1 might cause intrauterine adhesion.Conclusions At the same inoculation concentration,no significant difference is found in the pathogenicity between Up1 and Uu8,both of which appear to mainly cause cervicitis.Upl might be partially responsible for intrauterine adhesion in mice.