RESUMEN
BACKGROUND: Streptococcus mutans forms biofilms as a resistance mechanism against antimicrobial agents in the human oral cavity. We recently showed that human cathelicidin LL-37 exhibits inhibitory effects on biofilm formation of S. mutans through interaction with lipoteichoic acid (LTA), but without antibacterial or biofilm dispersal abilities. (-)-Epigallocatechin gallate (EGCG) is the most abundant constituent of tea catechins that has the greatest anti-infective potential to inhibit the growth of various microorganisms and biofilm formation. Therefore, in this study, we evaluated whether LL-37 interacts with EGCG to enhance the antibiofilm effect of EGCG on S. mutans biofilm formation. METHODS: Clinical S. mutans strains (n = 10) isolated from children's saliva were tested in a biofilm formation assay. The antibiofilm effect of EGCG with and without LL-37 was analyzed by the minimum biofilm eradication concentration assay and confirmed using field emission-scanning electron microscopy. In addition, the interaction among EGCG, LL-37, and LTA of S. mutans was determined using quartz crystal microbalance analysis. RESULTS: EGCG killed 100 % of planktonic S. mutans within 5 h, inhibited biofilm formation within 24 h, and reduced bacteria cells in preformed biofilms within 3 h at a concentration of 0.2 mg/mL. However, EGCG did not appear to interact with LTA. LL-37 effectively enhanced the bactericidal activity of EGCG against biofilm formation and preformed biofilms as determined by quantitative crystal violet staining and field emission-scanning electron microscopy. In addition, quartz crystal microbalance analysis revealed that LL-37 interacted with EGCG and promoted binding between EGCG and LTA of S. mutans. CONCLUSIONS: We show that LL-37 enhances the antibiofilm effect of EGCG on S. mutans. This finding provides new knowledge for dental treatment by using LL-37 as a potential antibiofilm compound.
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Antibacterianos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Biopelículas/efectos de los fármacos , Streptococcus mutans , Catequina/análogos & derivados , Catequina/farmacología , Humanos , Pruebas de Sensibilidad Microbiana , CatelicidinasRESUMEN
BACKGROUND: Enterovirus 71 (EV71) is one of the major etiological pathogens of hand, foot and mouth disease (HFMD) and can cause severe cerebral and pulmonary complications and even fatality. MicroRNAs (miRNAs), a class of small non-coding RNA molecules, play an important role in post-transcriptional regulation of gene expression and thereby influencing various physiological and pathological processes. Increasing evidence suggests that miRNAs act as key effector molecules in the complicated pathogen-host interactions. However, the roles of miRNAs in EV71 infection and pathogenesis are not well understood. METHODS: To identify special miRNAs involved in EV71 infection, a microarray assay was performed to study the expression pattern of miRNAs in EV71-infected human rhabdomyosarcoma cells (RD cells) and uninfected RD cells. We further predicted the putative target genes for the dysregulated miRNAs using the online bioinformatic algorithms (TargetScan, miRanda and PicTar) and carried out functional annotation including GO enrichment and KEGG pathway analysis for miRNA predicted targets. Then, the results of microarray were further confirmed by quantitative RT-PCR. RESULTS: Totally, 45 differentially expressed miRNAs ware identified by microarray, among which 36 miRNAs were up-regulated and 9 were down-regulated. 7166 predicted target genes for the dysregulated miRNAs were revealed by using TargetScan in conjunction with miRanda and PicTar. The GO annotation suggested that predicted targets of miRNAs were enriched into the category of signal transduction, regulation of transcription, metabolic process, protein phosphorylation, apoptotic process and immune response. KEGG pathway analysis suggested that these predicted target genes were involved in many important pathways, mainly including endocytosis and focal adhesion, MAPK signaling pathway, hypertrophic cardiomyopathy, melanogenesis and ErbB signaling pathway. The expression levels of 8 most differentially up-regulated miRNAs and 3 most differentially down-regulated miRNAs were confirmed by qRT-PCR. The expressions of hsa-miR-4530, hsa-miR-4492, hsa-miR-6125, hsa-miR-494-3p, hsa-miR-638, hsa-miR-6743-5p, hsa-miR-4459 and hsa-miR-4443 detected by qRT-PCR were consistent with the microarray data. CONCLUSION: These results might extend our understanding to the regulatory mechanism of miRNAs underlying the pathogenesis of EV71 infection, thus strengthening the preventative and therapeutic strategies of HFMD caused by EV71.
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Enterovirus Humano A/crecimiento & desarrollo , Interacciones Huésped-Patógeno , MicroARNs/biosíntesis , Línea Celular Tumoral , Biología Computacional , Perfilación de la Expresión Génica , Humanos , MicroARNs/genética , Análisis por Micromatrices , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Vibrio vulnificus (Vv) is known to cause life-threatening infections, particularly septicemia. These patients often exhibit elevated levels of pro-inflammatory cytokines. While it is established that mitogen-activated protein kinase (MAPK)-interacting kinase (MNK) contributes to the production of pro-inflammatory cytokines, the role of MNK in macrophages during Vv infection remains unclear. In this study, we investigate the impact of MNK on macrophages. We demonstrate that the inhibition of MNK in J774A.1 cells, when treated with lipopolysaccharide or Vv, resulted in decreased production of tumor necrosis factor alpha and interleukin-6, without affecting their transcription. Interestingly, treatment with MNK inhibitor CGP57380 led to enhanced phosphorylation of MNK1 but decreased phosphorylation of eIF4E. Moreover, MNK1 knockout cells exhibited an increased capacity for phagocytosis and clearance of Vv, with more acidic phagosomes than the parental cells. Notably, CGP57380 did not impact phagocytosis, bacterial clearance, or phagosome acidification in Vv-infected J774A.1 cells. Considering the reported association between MNK and mammalian target of rapamycin complex 1 (mTORC1) activation, we investigated the mTORC1 signaling in MNK1 knockout cells infected with Vv. Our results revealed that attenuation of the mTORC1 signaling in these cells and treatment with the mTORC1 inhibitor rapamycin significantly enhanced bacterial clearance in J774A.1 cells following Vv infection. In summary, our findings suggest that MNK promotes the Vv-induced cytokine production in J774A.1 cells without affecting their transcription levels. MNK1 appears to impair the phagocytosis, bacterial clearance, and phagosome acidification in Vv-infected J774A.1 cells through the MNK1-mTORC1 signaling pathway rather than the MNK1-eIF4E signaling pathway. Our findings highlight the importance of the MNK1-mTORC1 pathway in modulating macrophage responses to Vv infection. IMPORTANCE: Mitogen-activated protein kinase (MAPK)-interacting kinase (MNK) plays a role in promoting the production of tumor necrosis factor alpha and interleukin-6 in macrophages during Vibrio vulnificus (Vv) infection. Inhibition or knockout of MNK1 in J774A.1 cells resulted in reduced cytokine production without affecting their transcription levels. MNK1 also impairs phagocytosis, bacterial clearance, and phagosome acidification in Vv-infected cells through the MNK1-mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. The findings highlight the importance of the MNK1-mTORC1 pathway in modulating macrophage responses to Vv infection.
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Macrófagos , Diana Mecanicista del Complejo 1 de la Rapamicina , Fagocitosis , Proteínas Serina-Treonina Quinasas , Vibrio vulnificus , Vibrio vulnificus/metabolismo , Vibrio vulnificus/genética , Macrófagos/microbiología , Macrófagos/inmunología , Macrófagos/metabolismo , Animales , Ratones , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Línea Celular , Vibriosis/inmunología , Vibriosis/microbiología , Transducción de Señal , Citocinas/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Fosforilación , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Humanos , Compuestos de Anilina , PurinasRESUMEN
Leifsonia aquatica is an aquatic bacterium that is typically found in environmental water habitats. Infections due to L. aquatica are rare and commonly catheter associated in immunocompromised patients. We report the first case of an acute septicemia caused by L. aquatica in a healthy immunocompetent host after cryopexy in the absence of a catheter.
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Infecciones por Actinomycetales/diagnóstico , Actinomycetales/aislamiento & purificación , Complicaciones Posoperatorias/diagnóstico , Retina/cirugía , Sepsis/diagnóstico , Infecciones por Actinomycetales/microbiología , Infecciones por Actinomycetales/patología , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Complicaciones Posoperatorias/microbiología , Complicaciones Posoperatorias/patología , ARN Ribosómico 16S/genética , Sepsis/microbiología , Sepsis/patología , Análisis de Secuencia de ADNRESUMEN
The study for the first time attempted to accomplish 18S ribosomal DNA (rDNA) complete sequence amplification and analysis for three Demodex species (Demodex folliculorum, Demodex brevis and Demodex canis) based on gDNA extraction from individual mites. The mites were treated by DNA Release Additive and Hot Start II DNA Polymerase so as to promote mite disruption and increase PCR specificity. Determination of D. folliculorum gDNA showed that the gDNA yield reached the highest at 1 mite, tending to descend with the increase of mite number. The individual mite gDNA was successfully used for 18S rDNA fragment (about 900 bp) amplification examination. The alignments of 18S rDNA complete sequences of individual mite samples and those of pooled mite samples ( ≥ 1000mites/sample) showed over 97% identities for each species, indicating that the gDNA extracted from a single individual mite was as satisfactory as that from pooled mites for PCR amplification. Further pairwise sequence analyses showed that average divergence, genetic distance, transition/transversion or phylogenetic tree could not effectively identify the three Demodex species, largely due to the differentiation in the D. canis isolates. It can be concluded that the individual Demodex mite gDNA can satisfy the molecular study of Demodex. 18S rDNA complete sequence is suitable for interfamily identification in Cheyletoidea, but whether it is suitable for intrafamily identification cannot be confirmed until the ascertainment of the types of Demodex mites parasitizing in dogs.
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ADN Ribosómico/química , Genoma/genética , Ácaros/genética , ARN Ribosómico 18S/genética , Animales , Secuencia de Bases , Clonación Molecular , ADN Ribosómico/aislamiento & purificación , Ácaros/clasificación , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
In the present study, surveys of case numbers, constituent ratios, conventional biotyping, and multilocus sequence typing (MLST) were applied to characterize the incidence rate and epidemiological characteristics of human brucellosis in Shaanxi Province, China. A total of 12,215 human brucellosis cases were reported during 2008-2020, for an annual average incidence rate of 2.48/100,000. The most significant change was that the county numbers of reported cases increased from 36 in 2008 to 84 in 2020, with a geographic expansion trend from northern Shaanxi to Guanzhong, and southern Shaanxi regions; the incidence rate declined in previous epidemic northern Shaanxi regions while increasing each year in Guanzhong and southern Shaanxi regions such as Hancheng and Xianyang. The increased incidence was closely related to the development of large-scale small ruminants (goats and sheep) farms in Guanzhong and some southern Shaanxi regions. Another significant feature was that student cases (n = 261) were ranked second among all occupations, accounting for 2.14% of the total number of cases, with the majority due to drinking unsterilized goat milk. Three Brucella species were detected (B. melitensis (bv. 1, 2, 3 and variant), B. abortus bv. 3/6, and B. suis bv. 1) and were mainly distributed in the northern Shaanxi and Guanzhong regions. Three known STs (ST8, ST2, and ST14) were identified based on MLST analysis. The characteristics that had not changed were that B. melitensis strains belonging to the ST8 population were the dominant species and were observed in all nine regions during the examined periods. Strengthened human and animal brucellosis surveillance and restriction of the transfer of infected sheep (goats) as well as students avoiding drinking raw milk are suggested as optimal control strategies.
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Brucella melitensis/genética , Brucelosis/epidemiología , Monitoreo Epidemiológico , Tipificación de Secuencias Multilocus , Animales , China/epidemiología , Femenino , Variación Genética , Genotipo , Geografía , Cabras , Humanos , Incidencia , Masculino , Leche , OvinosRESUMEN
MicroRNAs (miRNAs), which are evolutionarily well-conserved, 21- to 23-nucleotide-long, small non-coding RNAs, are widely involved in the regulation of multiple biological processes, such as development, organogenesis, cell proliferation, cell differentiation, and apoptosis. Recent studies have shown that polymorphism in miRNAs and their target sites are closely related to various diseases, such as tumor and cardiological diseases. This review introduces recent progresses on polymorphism in microRNAs and their targeting sites and the related diseases.
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MicroARNs/genética , MicroARNs/metabolismo , Polimorfismo Genético , Metilación de ADN , Enfermedad/genética , Humanos , MutaciónRESUMEN
BACKGROUND: Preterm premature rupture of membrane (PPROM) can lead to serious consequences such as intrauterine infection, prolapse of the umbilical cord, and neonatal respiratory distress syndrome. Genital infection is a very important risk which closely related with PPROM. The preliminary study only made qualitative research on genital infection, but there was no deep and clear judgment about the effects of pathogenic bacteria. This study was to analyze the association of infections with PPROM in pregnant women in Shaanxi, China, and to establish Bayesian stepwise discriminant analysis to predict the incidence of PPROM. METHODS: In training group, the 112 pregnant women with PPROM were enrolled in the case subgroup, and 108 normal pregnant women in the control subgroup using an unmatched case-control method. The sociodemographic characteristics of these participants were collected by face-to-face interviews. Vaginal excretions from each participant were sampled at 28-36+6 weeks of pregnancy using a sterile swab. DNA corresponding to Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), Candida albicans, group B streptococci (GBS), herpes simplex virus-1 (HSV-1), and HSV-2 were detected in each participant by real-time polymerase chain reaction. A model of Bayesian discriminant analysis was established and then verified by a multicenter validation group that included 500 participants in the case subgroup and 500 participants in the control subgroup from five different hospitals in the Shaanxi province, respectively. RESULTS: The sociological characteristics were not significantly different between the case and control subgroups in both training and validation groups (all P > 0.05). In training group, the infection rates of UU (11.6% vs. 3.7%), CT (17.0% vs. 5.6%), and GBS (22.3% vs. 6.5%) showed statistically different between the case and control subgroups (all P < 0.05), log-transformed quantification of UU, CT, GBS, and HSV-2 showed statistically different between the case and control subgroups (P < 0.05). All etiological agents were introduced into the Bayesian stepwise discriminant model showed that UU, CT, and GBS infections were the main contributors to PPROM, with coefficients of 0.441, 3.347, and 4.126, respectively. The accuracy rates of the Bayesian stepwise discriminant analysis between the case and control subgroup were 84.1% and 86.8% in the training and validation groups, respectively. CONCLUSIONS: This study established a Bayesian stepwise discriminant model to predict the incidence of PPROM. The UU, CT, and GBS infections were discriminant factors for PPROM according to a Bayesian stepwise discriminant analysis. This model could provide a new method for the early predicting of PPROM in pregnant women.
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Teorema de Bayes , Rotura Prematura de Membranas Fetales/epidemiología , Rotura Prematura de Membranas Fetales/etiología , Adulto , Estudios de Casos y Controles , Análisis Discriminante , Femenino , Humanos , Embarazo , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de Riesgo , Adulto JovenRESUMEN
Pololike kinase 2 (PLK2) is a serine/threonine protein kinase, which has vital roles during mitosis and the centrosome cycle. In acute myeloblastic leukemia and hepatocarcinogenesis, PLK2 acts as a tumor suppressor; however, the function of PLK2 in gastric cancer remains to be elucidated. In the present study, PLK2 was overexpressed in gastric cancer tissues and three types of gastric cancer cells, SGC7901, MKN45 and BGC823. Transfection of SGC7901 gastric cancer cells with small interfering (si)RNA against PLK2 exerted no effect on the ratio of cells at different stages of the cell cycle compared with that of the untransfected and control siRNAtransfected cells. In addition, silencing of PLK2 significantly enhanced the growth of SGC7901 cells through inhibiting apoptosis. Furthermore, apoptosisassociated genes Bax and caspase 3 were found to be downregulated at the protein level. In conclusion, these results suggested that PLK2 may act as a tumor suppressor in gastric cancer, therefore indicating its therapeutic potential.
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Apoptosis/genética , Silenciador del Gen , Proteínas Serina-Treonina Quinasas/genética , Neoplasias Gástricas/genética , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Ciclo Celular/genética , Línea Celular Tumoral , Proliferación Celular , Expresión Génica , Humanos , Interferencia de ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genéticaAsunto(s)
Arvicolinae , Infecciones por Bartonella/veterinaria , Bartonella/aislamiento & purificación , Lagomorpha , Muridae , Enfermedades de los Roedores/epidemiología , Roedores , Animales , Proteínas Bacterianas/análisis , Bartonella/clasificación , Infecciones por Bartonella/epidemiología , Infecciones por Bartonella/microbiología , China/epidemiología , ADN Bacteriano/análisis , Filogenia , Prevalencia , Enfermedades de los Roedores/microbiología , Análisis de Secuencia de ADN/veterinariaRESUMEN
MicroRNA (miRNA)-126 (miR-126) was reported to be downregulated and to act as a tumor suppressor in cancers of the lung, cervix, bladder and prostate. However, the functions of miR-126 in gastric cancer appear to be diverse and are largely unknown. MiR-126 was reported to act as a tumor suppressor by targeting the Crk gene, or as an oncogene by targeting the SOX2 gene in gastric cancer. We identified that the expression of miR-126 was decreased in gastric cancer cell lines and tissues. PLK2, a tumor suppressor gene, was directly regulated by miR-126 in SGC-7901 cells. Overexpression of miR-126 not only suppressed the growth and clone formation of SGC-7901 cells, but also induced apoptosis in vitro, whereas inhibition of miR-126 slightly promoted SGC-7901 cell proliferation. The cell cycle was not affected by miR-126. Moreover, miR-126 suppressed tumor growth in vivo in a xenograft model. PLK2, PI3KR2 and Crk were regulated by miR-126 in SGC-7901 cells. We infer that the functions of miR-126 in gastric cancer depend on synergistic targeting balance between oncogenes and anti-oncogenes. Our study indicates that miR-126 is a tumor suppressor, which in the future may become a therapeutic target for gastric cancer.
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Fosfatidilinositol 3-Quinasa Clase Ia/metabolismo , MicroARNs/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-crk/metabolismo , Neoplasias Gástricas/patología , Animales , Línea Celular Tumoral , Fosfatidilinositol 3-Quinasa Clase Ia/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Ratones , Ratones Desnudos , MicroARNs/genética , Neoplasias Experimentales , Proteínas Serina-Treonina Quinasas/genética , Proteínas Proto-Oncogénicas c-crk/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismoRESUMEN
PURPOSE: To conduct a meta-analysis to confirm the association between Demodex infestation and blepharitis. METHODS: We conducted a comprehensive and quantitative analysis of relevant published case-control studies which were found from the ISI Web of Knowledge, MEDLINE and CNKI from January 1950 to December 2010. Meta-analysis was applied for 13 of these and included matched data sets, using odds ratio (OR) as the effect indicator. Sensitivity was assessed. RESULTS: Eleven articles (13 matched data sets) covering four different countries and reporting 4741 participants (2098 blepharitis and 2643 controls) were eligible. The pooled OR in random effect models was 4.89 (95% confidence interval, 3.00-7.97). Sensitivity analysis showed that results of pooled ORs in different effect models, language, sample size, and control groups were completely consistent, which demonstrated a stable association between Demodex infestation and blepharitis by meta-analysis. CONCLUSIONS: The association between Demodex infestation and blepharitis was statistically significant. The conclusion implies that when conventional treatments for blepharitis fail, examination of Demodex mites and acaricidal therapy should be considered.
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Blefaritis/parasitología , Infecciones Parasitarias del Ojo/parasitología , Infestaciones por Ácaros/parasitología , Ácaros , Animales , Blefaritis/epidemiología , Estudios de Casos y Controles , Infecciones Parasitarias del Ojo/epidemiología , Humanos , Infestaciones por Ácaros/epidemiología , Oportunidad RelativaRESUMEN
To our knowledge, few reports on Demodex studied at the molecular level are available at present. In this study our group, for the first time, cloned, sequenced and analyzed the chitin synthase (CHS) gene fragments of Demodex folliculorum, Demodex brevis, and Demodex canis (three isolates from each species) from Xi'an China, by designing specific primers based on the only partial sequence of the CHS gene of D. canis from Japan, retrieved from GenBank. Results show that amplification was successful only in three D. canis isolates and one D. brevis isolate out of the nine Demodex isolates. The obtained fragments were sequenced to be 339 bp for D. canis and 338 bp for D. brevis. The CHS gene sequence similarities between the three Xi'an D. canis isolates and one Japanese D. canis isolate ranged from 99.7% to 100.0%, and those between four D. canis isolates and one D. brevis isolate were 99.1%-99.4%. Phylogenetic trees based on maximum parsimony (MP) and maximum likelihood (ML) methods shared the same clusters, according with the traditional classification. Two open reading frames (ORFs) were identified in each CHS gene sequenced, and their corresponding amino acid sequences were located at the catalytic domain. The relatively conserved sequences could be deduced to be a CHS class A gene, which is associated with chitin synthesis in the integument of Demodex mites.
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Quitina Sintasa/genética , Ácaros/enzimología , Ácaros/genética , Animales , Secuencia de Bases , Clonación Molecular , Cartilla de ADN/genética , Ácaros/clasificación , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Filogenia , Homología de Secuencia de Ácido Nucleico , Especificidad de la EspecieRESUMEN
To identify sociodemographic characteristics and risk factor of Demodex infestation, 756 students aged 13-22 years in Xi'an, China were sampled for the school-based cross-sectional study. Demodex was examined using the cellophane tape method (CTP). The results showed that the total detection rate of Demodex was 67.6%. Logistic regression analysis revealed that five variables (gender, residence, sharing sanitary ware, frequency of face-wash per day, and use of facial cleanser) were found to be uncorrelated with Demodex infestation, whereas three variables (age, skin type, and skin disease) were found to be independent correlates. Students aged over 18 years had 22.1 times higher odds of Demodex infestation compared to those under 16 years and students aged 16-18 years also had 2.1 times higher odds compared to those aged 13-15 years. Odds of having a Demodex infestation for oily or mixed skin were 2.1 times those for dry or neutral skin. Students with a facial skin disease had 3.0 times higher odds of being infested with Demodex compared to those without. The inception rate of students with facial dermatoses increased in parallel with increasing mite count. The inception rates were 21.3%, 40.7%, 59.2%, and 67.7% in the negative, mild, moderate, and severe infestation groups, respectively (χ(2)=60.6, P<0.001). Specifically, the amount of infested mites and inception rate of acne vulgaris were positively correlated (R(2)=0.57, moderate infestation odds ratio (OR)=7.1, severe infestation OR=10.3). It was concluded that Demodex prevalence increases with age, and Demodex presents in nearly all adult human. Sebaceous hyperplasia with oily or mixed skin seems to favour Demodex proliferation. Demodex infestation could be associated with acne vulgaris. The CTP is a good sampling method for studies of Demodex prevalence.