The diagnostic value of serum levels of pro calcitonin, β2 defensins, C-reactive protein and the positive rate of group B streptococci in preterm premature rupture of membranes with amniotic infection / 中国医师杂志

Objective:To explore the diagnostic value of serum levels of pro calcitonin (PCT), β2 defensins (HBD-2), C-reactive protein (CRP) and the positive rate of group B streptococci (GBS) in preterm premature rupture of membranes (PROM) with amniotic infection.Methods:This study was a retrospective study. 156 pregnant women with preterm PROM who were diagnosed by the Obstetrics Department of the Hospital of Southern University of Science and Technology from January 2017 to January 2022 were selected as the study subjects. According to whether there was amniotic infection, they were divided into 57 infected women and 99 non infected women. The levels of serum PCT, HBD-2 and CRP before delivery were compared between the two groups, and the positive rate of GBS in vaginal discharge was detected, and the receiver operating curve (ROC) was used to analyze the value of various indicators in diagnosing amniotic cavity infection in preterm PROM mothers.Results:The serum levels of PCT, HBD-2, CRP, and GBS positivity in the infected group were significantly higher than those in the non infected group, with statistically significant differences (all P<0.01); The area under the curve (AUC) value, sensitivity, and specificity of serum PCT for diagnosing preterm PROM with amniotic cavity infection were 0.894, 82.56%, and 80.74%, respectively; The AUC value of HBD-2 for diagnosing preterm PROM with amniotic cavity infection was 0.792, the sensitivity was 70.78%, and the specificity was 77.59%; The AUC value, sensitivity, and specificity of CRP in diagnosing preterm PROM with amniotic cavity infection were 0.756, 68.94%, and 72.78%, respectively; The positive rate of GBS in vaginal discharge was 0.733, the sensitivity was 64.91%, and the specificity was 81.82%. Conclusions:The serum levels of PCT, HBD-2, CRP and the positive rate of GBS in vaginal discharge of pregnant women with preterm PROM complicated with amniotic infection will increase significantly. All indicators have high practical value for the diagnosis of preterm PROM complicated with amniotic infection.

β-defensinas como posibles indicadores de la actividad inflamatoria en la enfermedad periodontal / β-defensins as possible indicators of inflammatory activity in periodontal disease

Resumen La enfermedad periodontal (gingivitis y periodontitis) es un proceso inflamatorio ocasionado por la actividad de bacterias patógenas y sus productos sobre el surco gingival, con la consecuente activación de la respuesta inmunitaria. La saliva y el fluido crevicular contienen una gran variedad de enzimas y factores antimicrobianos que están en contacto con la región supragingival y subgingival; entre ellos, las β-defensinas (hBDs). Las hBDs son péptidos catiónicos no glicosilados ricos en cisteína, producidos por las células epiteliales; tienen efecto antimicrobiano e inmunorregulador; de esta forma, contribuyen al mantenimiento de la homeostasis en los tejidos periodontales. Los cambios en la microbiota y en la respuesta inmunitaria de un periodonto sano a gingivitis y, finalmente, a periodontitis, es compleja. Su severidad depende de un equilibrio dinámico entre las bacterias asociadas a la placa, factores genéticos y ambientales. Los avances recientes han permitido comprender la implicación de las hBDs en la detección, el diagnóstico y la terapéutica de la enfermedad periodontal, así como la relación que hay entre la periodontitis y otras enfermedades inflamatorias. El objetivo de esta revisión es describir el efecto de las hBDs en la respuesta inmunitaria y su utilización como marcadores de la actividad inflamatoria de la enfermedad periodontal.

Abstract Periodontal disease (gingivitis and periodontitis) is an inflammatory process caused by the activity of pathogenic bacteria and their products on the gingival sulcus, with the consequent activation of the immune response. Saliva and crevicular fluid contain a wide variety of enzymes and antimicrobial factors that are in contact with the supragingival and subgingival region, including β-defensins (hBDs). hHBDs are non-glycosylated, cysteine-rich cationic peptides produced by epithelial cells with antimicrobial and immunoregulatory effects, thus contributing to maintaining homeostasis in periodontal tissues. The changes in the microbiota and the immune response from a healthy periodontium to gingivitis and, finally, to periodontitis are complex. Their severity depends on a dynamic balance between bacteria associated with plaque, genetic and environmental factors. Recent advances have made it possible to understand the implication of hBDs in the detection, diagnosis, and therapy of periodontal disease and the relationship between periodontitis and other inflammatory conditions. This review aims to describe the effect of hBDs on the immune response and its use as a possible marker of the inflammatory activity of the periodontal disease.

Potential anticancer peptides design from the cysteine rich plant defensins: An in silico approach-Supplementary Data

Cancer is the second leading cause of mortality worldwide preceded by cardiovascular diseases. The therapeutic approaches for drug developmentinclude the use of small molecules, antibodies, peptidesor short nucleic acid sequences. The peptide-based drugs have been developed to treat many diseases like cardiovascular diseases, cancer, metabolic disorders, immunological diseases and viral infections. More than 80 peptide drugs are already in the market. These therapeutic peptides have several important benefits over antibodies and proteins due to their small size, ease for chemical synthesis and further the ability to penetrate cell membrane. Furthermore, peptide drugs have high specificity, activity, and affinity. The plant defensins BcDef1, TPP3, NaD1, 2N2R and 2LR3 have been studied for their role in wide range of diseases. This study focussed on the conformation of plant defensins rich in disulfide bonds. The structure for BcDef1 has been predicted from the conformational ensemble. Then, we designed anticancer peptides from these defensins with computational methods. The designed anticancer peptides have been studied for their immunogenicity as well as homology with human proteome. The role of designed peptides has been suggested for interferon-gamma induction, the later has been shown to possess a very important role in cancer.

Potential anticancer peptides design from the cysteine rich plant defensins: An in silico approach-Supplementary Data

Cancer is the second leading cause of mortality worldwide preceded by cardiovascular diseases. The therapeutic approaches for drug developmentinclude the use of small molecules, antibodies, peptidesor short nucleic acid sequences. The peptide-based drugs have been developed to treat many diseases like cardiovascular diseases, cancer, metabolic disorders, immunological diseases and viral infections. More than 80 peptide drugs are already in the market. These therapeutic peptides have several important benefits over antibodies and proteins due to their small size, ease for chemical synthesis and further the ability to penetrate cell membrane. Furthermore, peptide drugs have high specificity, activity, and affinity. The plant defensins BcDef1, TPP3, NaD1, 2N2R and 2LR3 have been studied for their role in wide range of diseases. This study focussed on the conformation of plant defensins rich in disulfide bonds. The structure for BcDef1 has been predicted from the conformational ensemble. Then, we designed anticancer peptides from these defensins with computational methods. The designed anticancer peptides have been studied for their immunogenicity as well as homology with human proteome. The role of designed peptides has been suggested for interferon-gamma induction, the later has been shown to possess a very important role in cancer.

Potential anticancer peptides design from the cysteine rich plant defensins: An in silico approach

Cancer is the second leading cause of mortality worldwide preceded by cardiovascular diseases. The therapeutic approaches for drug developmentinclude the use of small molecules, antibodies, peptidesor short nucleic acid sequences. The peptide-based drugs have been developed to treat many diseases like cardiovascular diseases, cancer, metabolic disorders, immunological diseases and viral infections. More than 80 peptide drugs are already in the market. These therapeutic peptides have several important benefits over antibodies and proteins due to their small size, ease for chemical synthesis and further the ability to penetrate cell membrane. Furthermore, peptide drugs have high specificity, activity, and affinity. The plant defensins BcDef1, TPP3, NaD1, 2N2R and 2LR3 have been studied for their role in wide range of diseases. This study focussed on the conformation of plant defensins rich in disulfide bonds. The structure for BcDef1 has been predicted from the conformational ensemble. Then, we designed anticancer peptides from these defensins with computational methods. The designed anticancer peptides have been studied for their immunogenicity as well as homology with human proteome. The role of designed peptides has been suggested for interferon-gamma induction, the later has been shown to possess a very important role in cancer.

Impact of smoking on protein levels of beta-defensins in periodontal disease

Abstract Antimicrobial peptides (AMPs) are important components of the host response against invading pathogens. In addition to their direct antimicrobial activity, they can also participate in the immune system modulation. However, the role of AMPs in the etiopathogenesis of periodontal disease and the risk factors that may influence their expression in the oral cavity are not fully understood. The aim of this study was to determine the impact of smoking on beta-defensin (hBD) 1 and 2 levels analyzing samples from periodontitis patients. Fifty patients with periodontitis, 25 smokers and 25 non-smokers, and 20 periodontally healthy patients were recruited. After periodontal clinical evaluation, gingival crevicular fluid (GCF) samples were collected from healthy sites of patients without periodontal disease and from healthy and diseased sites of patients with periodontitis. Peptides quantification was performed by sandwich ELISA technique. Smokers showed reduced GCF hBD 1 levels and increased hBD 2 levels compared to non-smokers in diseased sites (p <0.05). Higher levels of hBD 1 were observed in healthy sites of patients without periodontal disease than in healthy sites of patients with periodontitis (p<0.0001). Diseased sites of non-smokers presented higher levels of hBD 2 than healthy sites (p <0.05). These results reveal that protein levels of hBDs 1 and 2 can be impaired by cigarette smoking in the presence of periodontal disease.

Resumo Peptídeos antimicrobianos (PAMs) são componentes importantes da resposta do hospedeiro contra patógenos invasores. Além de sua atividade antimicrobiana direta, eles também podem participar da modulação do sistema imunológico. No entanto, o papel dos PAMs na etiopatogenia da doença periodontal e os fatores de risco que podem influenciar a sua expressão na cavidade oral não são totalmente compreendidos. O objetivo deste estudo foi determinar o impacto do tabagismo nos níveis de beta-defensina (hBD) 1 e 2 analisando amostras de pacientes com periodontite. Cinquenta pacientes com periodontite, 25 fumantes e 25 não fumantes e 20 pacientes periodontalmente saudáveis foram recrutados. Após avaliação clínica periodontal, amostras de fluido crevicular gengival (FCG) foram coletadas de sítios saudáveis de pacientes sem doença periodontal e de sítios saudáveis e doentes de pacientes com periodontite. A quantificação dos peptídeos foi realizada pela técnica de ELISA sanduíche. Fumantes apresentaram níveis reduzidos de hBD 1 no FCG e níveis aumentados de hBD 2 em comparação com não fumantes em locais doentes (p <0,05). Níveis mais elevados de hBD 1 foram observados em sítios saudáveis de pacientes sem doença periodontal do que em sítios saudáveis de pacientes com periodontite (p<0,0001). Os sítios doentes de não fumantes apresentaram níveis mais elevados de hBD 2 do que os sítios saudáveis (p<0,05). Esses resultados revelam que os níveis das hBDs 1 e 2 podem ser prejudicados pelo tabagismo na presença de doença periodontal.

Defensin c-thionin from Capsicum chinense improves butyrate cytotoxicity on human colon adenocarcinoma cell line Caco-2

BACKGROUND: Butyrate is a histone deacetylase inhibitor that induces apoptosis and inhibits cell proliferation of colorectal cancer cells. To improve its anticancer activity, butyrate has been evaluated mixed with drugs and different molecules. Plant antimicrobial peptides are attractive anticancer alternative molecules because they show selective cytotoxic activity against different cancer cell lines. In this work, we explore if the plant defensin c-thionin (Capsicum chinense) can improve butyrate activity on Caco-2 cell line and we also determined the mechanism of death activated. RESULTS: The combined treatment of c-thionin (3.5 mM) and butyrate (50 mM) showed higher cytotoxicity on Caco-2 cells with respect to single treatments. Also, the combined treatment reduced cell proliferation and exhibited a higher rate of apoptosis than single treatments. Combined treatment induced caspases 8 and 9 activation to an extent comparable with that of butyrate while c-thionin did not activate caspases. Additionally, reactive oxygen species generation preceded the onset of apoptosis, and superoxide anion production was higher in cells treated with the combined treatment. CONCLUSIONS: The c-thionin from Habanero chili pepper improved the butyrate cytotoxicity on Caco-2 cells. This effect occurred through apoptosis induction associated with reactive oxygen species production. Therefore, the combination of butyrate with cytotoxic antimicrobial peptides could be an attractive strategy for cancer therapy.

Inhibition of defensin A and cecropin A responses to dengue virus 1 infection in Aedes aegypti / Inhibición de las respuestas de defensina A y cecropina A contra la infección del virus dengue 1 en Aedes aegypti

Abstract | Introduction: It is essential to determine the interactions between viruses and mosquitoes to diminish dengue viral transmission. These interactions constitute a very complex system of highly regulated pathways known as the innate immune system of the mosquito, which produces antimicrobial peptides that act as effector molecules against bacterial and fungal infections. There is less information about such effects on virus infections. Objective: To determine the expression of two antimicrobial peptide genes, defensin A and cecropin A, in Aedes aegypti mosquitoes infected with DENV-1. Materials and methods: We used the F1 generation of mosquitoes orally infected with DENV-1 and real-time PCR analysis to determine whether the defensin A and cecropin A genes played a role in controlling DENV-1 replication in Ae. aegypti. As a reference, we conducted similar experiments with the bacteria Escherichia coli. Results: Basal levels of defensin A and cecropin A mRNA were expressed in uninfected mosquitoes at different times post-blood feeding. The infected mosquitoes experienced reduced expression of these mRNA by at least eightfold when compared to uninfected control mosquitoes at all times post-infection. In contrast with the behavior of DENV-1, results showed that bacterial infection produced up-regulation of defensin and cecropin genes; however, the induction of transcripts occurred at later times (15 days). Conclusion: DENV-1 virus inhibited the expression of defensin A and cecropin A genes in a wild Ae. aegypti population from Venezuela.

Resumen | Introducción. Es esencial determinar las interacciones entre los virus y los mosquitos para disminuir la transmisión viral. Estas interacciones constituyen un sistema muy complejo y muy regulado conocido como sistema inmunitario innato del mosquito, el cual produce péptidos antimicrobianos, moléculas efectoras que funcionan contra las infecciones bacterianas y fúngicas; se tiene poca información de su acción sobre los virus. Objetivo. Determinar la expresión de dos genes AMP (defensina A y cecropina A) en mosquitos Aedes aegypti infectados con el virus DENV-1. Materiales y métodos. Se infectaron oralmente mosquitos de generación F1 con DENV-1 y mediante el análisis con PCR en tiempo real se determinó el potencial papel de los genes defensina A y cecropina A en el control de la replicación del DENV-1 en Ae. aegypti. Como referencia, se infectaron mosquitos con Escherichia coli. Resultados: Los mosquitos no infectados expresaron niveles basales de los ARNm de los genes defensina A y cecropina A en diversos momentos después de la alimentación. Los mosquitos infectados experimentaron una reducción, por lo menos, de ocho veces en la expresión de estos ARNm con respecto a los mosquitos de control en todo el periodo posterior a la alimentación. En contraste con el comportamiento del virus DENV-1, los resultados mostraron que la infección bacteriana produjo una regulación positiva de los genes defensina y cecropina; sin embargo, la inducción de los transcritos ocurrió tardíamente (15 días). Conclusión. El virus DENV-1 inhibió la expresión de los genes defensina A y cecropina A en una población silvestre de Ae. aegypti en Venezuela.

The level of antimicrobial peptides in gingival crevicular fluid and its correlation with periodontal clinical indexes in elderly patients with type 2 diabetic periodontitis / 口腔疾病防治

Objective@#To investigate the changes and significance of human beta-defensin-2 (HBD-2) and LL-37 in the gingival crevicular fluid of patients with periodontitis and type 2 diabetes mellitus (T2DM).@*Methods@#This study was conducted among 45- to 85-year-old patients in the Department of Stomatology and Internal Medicine of Shenzhen Center for Chronic Disease Control, including a healthy control group of 22 people, a systemically healthy control group of 19 people with periodontitis, a T2DM periodontal health group of 15 people, and a T2DM group of 21 people with periodontitis. The Florida periodontal probe was used for periodontal examination, and the clinical indexes, including probing depth (PD), clinical attachment level (CAL) and probing on bleeding (BOP), were recorded. The concentrations of HBD-2 and Ll-37 in gingival crevicular fluid were determined by ELISA. The differences in HBD-2, LL-37 and periodontal clinical indexes between the groups were compared, and correlation analysis was conducted.@*Results@#The PD values in T2DM with the periodontitis group were higher than those of the systemically healthy controls with periodontitis group (P < 0.05); the levels of HBD-2 and LL-37 in gingival crevicular fluid in systemically healthy controls with periodontitis group were significantly higher than those in the healthy control group (P < 0.05), the level of HBD-2 in gingival crevicular fluid in systemically healthy controls with periodontitis group was significantly higher than that in T2DM with periodontitis group (P < 0.05); and the antimicrobial peptides HBD-2 and LL-37 in gingival crevicular fluid were significantly positively correlated with the PD and CAL in systemically healthy controls with periodontitis group (P < 0.05), and there was no significant correlation between the antimicrobial peptides HBD-2, LL-37 in gingival crevicular fluid and PD, CAL in T2DM with periodontitis group (P > 0.05).@*Conclusion@#The levels of antimicrobial peptides HBD-2 and LL-37 in gingival crevicular fluid of middle-aged and elderly patients with T2DM periodontitis were lower, and there was no significant correlation with PD and CAL in periodontal clinical indicators.

Quantification of Beta-Defensins (DEFB) Gene Copy Number Variations in Relation to Inflammation in Type 2 Diabetes Mellitus and Diabetic Nephropathy Patients

@#Introduction: Association studies between single nucleotide polymorphisms (SNPs) and type 2 diabetes mellitus (T2DM) have been abundant. However, there are limited reports on copy number variations (CNVs) of beta-defensins (DEFB) gene in relation to T2DM. In this study, DEFB copy numbers were quantified in T2DM with nephropathy, T2DM without nephropathy and non-diabetic control groups to investigate its influence in chronic inflammation in Malaysian individuals. Methods: DEFB copy number in Malaysian individuals were quantified by using paralogue ratio tests (PRT) which allow direct quantification of gene copy number by using PRT107A and HSPD21 PRT primers. The copy number generated was then validated from insertion/deletion ratio measurement 5DEL (rs5889219) and two microsatellite analyses (EPEV-1 and EPEV-3). Results: DEFB copy number was found extending from 2 to 8 copies in the non-diabetic group (n=146), while in T2DM group (n=392), copy numbers were more extensive, ranging between 1 and 12 copies; with 1, 10 and 12 copies detected in T2DM with nephropathy group (n=202). Statistically, there is no significant difference in DEFB copy number between T2DM and the non-diabetic group (p=0.209) as well as between diabetic nephropathy and without nephropathy of the T2DM group (p=0.522). However, significant white blood cell (WBC) count was found between T2DM groups with and without diabetic nephropathy (p=0.000). Conclusion: Extreme DEFB copy numbers in T2DM with nephropathy group suggest future studies with bigger sample size are necessary to elucidate the true impact of CNVs of DEFB gene in promoting early onset of nephropathy in T2DM.

Associação entre beta-defensinas e periodontite / Association between beta-defensins and periodontitis: literature review

Objetivo: as beta-defensinas humanas (hBDs) podem ter um papel-chave na susceptibilidade às doenças na cavidade bucal. Além do efeito antimicrobiano direto, as hBDs aumentam a imunidade adaptativa. O objetivo deste estudo foi realizar uma revisão de literatura científica sobre a relação entre beta-defensinas (hBD) e periodontite. Material e métodos: foi realizada uma pesquisa bibliográfica na base de dados PubMed sobre a expressão de hBDs em indivíduos com periodontite. Os termos beta defensins e periodontitis foram utilizados nessa busca. Resultados: foram selecionados, por um revisor, sete artigos para serem incluídos nessa revisão de literatura: dois estudos de intervenção e cinco estudos transversais. Conclusão: o número de estudos sobre a expressão de beta-defensinas em indivíduos com periodontite é reduzido. O conhecimento sobre o papel das beta-defensinas na periodontite pode trazer um maior entendimento de sua etiopatogenia, além de possibilitar novos indicadores de risco e terapias. Estudos adicionais são necessários para a elucidação da relação entre esses peptídeos antimicrobianos e a periodontite.

Objective: human beta-defensins (hBDs) may play a key role in the susceptibility to diseases in the oral cavity. In addition to the direct antimicrobial effect, hBDs enhance adaptive immunity. The objective of this study was to investigate the literature on the relationship between hBD and periodontitis. Material and methods: a literature review was conducted in the PubMed database on the expression of hBDs in subjects with periodontitis. The terms "beta-defensins" and "periodontitis" were used in this search. Results: seven articles were selected being: two intervention studies and fi ve cross-sectional studies. Conclusion: the number of studies on the expression of beta-defensins in individuals with periodontitis is reduced. Knowledge about the role of beta-defensins in periodontitis may lead to a better understanding of their etiopathogenesis, in addition to providing new risk indicators and therapies. Additional studies are needed to elucidate the relationship between these antimicrobial peptides and periodontitis.

Feasibility of developing HAP risk warning model in critically ill patients based on genomic copy number polymorphisms of DEFA1/DEFA3 / 中华麻醉学杂志

Objective To evaluate the feasibility of developing hospital acquired pneumonia (HAP) risk warning model in critically ill patients based on genomic copy number polymorphisms (CNPs) of the genes encoding human neutrophil peptides 1-3 (DEFA1/DEFA3).Methods Seventy-seven HAP patients (group HAP) and 109 non-HAP patients of matched age and sex in intensive care unit (ICU) (group NHAP) were enrolled in the study.The genomic CNPs of DEFA1/DEFA3 was determined by realtime quantitative polymerase chain reaction after extracting DNA from peripheral blood samples.The source of patients,condition of endotracheal intubation within 24 h after admission to ICU,Acute Physiology Score,Acute Physiology and Chronic Health Evaluation Ⅱ score,Sequential Organ Failure Assessment score,mechanical ventilation time,length of hospital and ICU stay and outcomes were obtained.The predictive model was developed using logistic regression through combining DEFA1/DEFA3 copy numbers and clinical characteristics (Acute Physiology Score and source of emergency) within 24 h after admission to ICU.The receiver operating characteristic curve was used to evaluate the predictive efficacy of the model.Results The copy numbers of DEFA1/DEFA3 were significantly lower in HAP group than in NHAP group (P ＜0.05).The area under the receiver operating characteristic curve of the predictive model developed through combining the DEFA1/DEFA3 copy numbers with clinical characteristics was 0.789 (95％ CI 0.724-0.854) when the model was used for predicting HAP.Conclusion CNPs of DEFA1/DEFA3 can be used to develop the HAP risk warning model in critically ill patients.

An investigation of human beta-defensins and cathelicidin expression in patients with pterygium / Uma investigação da expressão de beta-defensinas humanas e de catelidicina em pacientes com pterígio

ABSTRACT Purpose: To investigate human beta-defensins (HBDs) and cathelicidin LL-37 (LL-37) expressions in patients with pterygium. Methods: In this retrospective consecutive case series, 26 pterygium specimens and 15 normal conjunctival specimens of 15 control subjects were in vestigated. Expressions of HBD-1, HBD-2, HBD-3, and LL-37 were assessed using immuno histochemical staining. A brown color in the cytoplasm and/or nuclei of epithelial cells indicated positive staining for HBDs and LL-37. For each antibody, the intensity of the reaction (negative [-], weak [1+], moderate [2+], or strong [3+]) was determined to describe the immunoreactions. Results: The median age was 52 years in both groups. There were no significant differences in age and sex between the groups (p=0.583, p=0.355, respectively). Of the 26 pterygium specimens, 15 (57.7%) (14 weak, 1 moderate staining) showed HBD-2 expression, which was not observed in any of the control specimens. One (3.8%) pterygium and one (6.7%) control specimen demonstrated weak staining for HBD-3. HBD-2 expression was significantly higher in the pterygium specimens than in the controls (p=0.002). None of the tissue specimens had positive staining for HBD-1 or LL-37 in either group (both; p=1.00). Conclusions: HBD-2 expression was higher in pterygium specimens than in the controls. HBD-2 expression that might be stimulated by inflammatory cytokines may be related to inflammation and fibrovascular proliferation and may play a role in pterygium pathogenesis.

RESUMO Objetivo: Investigar as expressões beta defensinas humanas (HBD) e catelicidina em pacientes com pterígio. Métodos: Nesta série de casos retrospectivos consecutivos, 26 espécimes de pterígio e 15 espécimes conjuntivais normais de 15 indivíduos controle foram investigados. As expressões de HBD-1, HBD-2, HBD-3 e catelicidina (LL-37) foram avaliadas por coloração imuno-histoquímica. Uma cor castanha no citoplasma ou nos núcleos de células epiteliais foi definida como coloração positiva para HBDs e LL-37. Para cada anticorpo foi determinada a intensidade da reação (negativo [-], fraco [1+], moderado [2+] ou forte [3+]) para descrever as imunoreações. Resultados: A idade média foi de 52 anos em ambos os grupos. Não houve diferença significativa entre os grupos em termos de idade e sexo (p=0,583, p=0,355, respectivamente). Das 26 amostras de pterígio, 15 (57,7%) (14 fracas e 1 moderada) demonstraram a expressão de HBD-2 enquanto não foi encontrada em nenhum dos espécimes de controlo. Um dos pterígios (3,8%) e um dos espécimes de controlo (6,7%) demonstraram fraca coloração para HBD-3. A expressão de HBD-2 foi significati vamente maior nos espécimes de pterígio do que nos controles (p=0,002). Nenhum dos espécimes de tecido apresentou coloração positiva para HBD-1 ou LL-37 em ambos os grupos (ambos p=1,00). Conclusão: Encontramos aumento da expressão de HBD-2 em espécimes de pte rígio em relação aos controles. A expressão de HBD-2 que pode ser estimulada por citocinas inflamatórias pode estar relacionada com inflamação e proliferação fibrovascular e pode desempenhar um papel na patogênese do pterígio.

Study of anti-inflammatory function of human bea defensin 2 on rats' acute rhinosinusitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To study the antiinflammatory function of human beta defensin 2(hβD-2) on acute rhinosinusitis in rats,in order to provide a new therapy for acute rhinosinusitis.METHODS Acute rhinosinusitis model were established on SD rats before and after the transfection of plasmid,the rats in experimental group were dropped with recombinant hβD-2 plasmid mixture in nose,while with empty plasmid mixture in control group.Immunohistochemistry method was used to prove the transfection results,nasal mucosa were hematoxylineosin stained to compare the pathological difference of nasal mucosa,nasal lavage fliud was collected and cultured to compare the colony number of the bacteria.RESULTS The expression of hβD-2 was confirmed by immunohistochemistry method,which mainly distributed in mucosal epithelium and gland,pathological results showed that the inflammation of nasal mucosa in experimental group was significantly relieved than that in control group.The number of Staphylococcus auresus colony number was significantly decreased in experimental group,while there was no significantly change in the control group.CONCLUSION Recombinant hβD-2 plasmid can be successfully transfected into the nasal mucosa of rats and expressed effectively.The anti-inflammatory ability of nasal mucosa was increased after the transfection,which is expected to provide a new treatment approach for acute rhinosinusitis.

TLR4/NF-κB signal pathway mediate HBD-2 in patients with preterm premature rupture of membrane associated with histologic chorioamnionitis / 中国综合临床

Objective To observe the effect of human β-defensins-2(HBD-2) for chorioamnionitis(HCA) pregnant women before term premature rupture of fetal membrane(PROM) process,and explore toll-like receptor 4 / nuclear factor-κ B (TLR4 / NF-κB) predominate role in the process of signal transduction pathway in the mechanism.Methods Fifty five women with PROM were enrolled in the study.According to the Results of pathological diagnosis of membranes,pregnant women with PROM divided into histological chorioamnionitis,HCA and non-HCA.The same sample without PROM pregnancies matching the same gestational ages were recruited as control group.We examined the messenger RNA(mRNA) of TLR4,NF-κB p65 and HBD-2 in placenta and fetal membrane real-time reverse transcription polymerase chain reactions by dsDNA-binding dyes of SYBR Green.Results (1)In the placenta,the level of TLR-4(17.15±4.52),NF-κB p65(47.11±14.23),HBD-2mRNA(27.35±2.67) in PROM group were significantly higher than the level of TLR-4(7.21±3.25),NF-κB p65(30.51±13.05),HBD-2mRNA(13.55±0.8) in control group(t=-1.966,-1.474,-1.754,P0.05).Conclusion Linear positive correlation of TLR4,NF-κB and HBD2 indicated that TLR4/NF-κB/HBD2 signal transduction pathway may be involved in the development of preterm premature rupture of membrane associated with histologic chorioamnionitis.

Construction and identification of recombinant lentivirus carrying human beta defensins-3 and connective tissue growth factor gene / 军事医学

Objective To construct a recombinant lentivirus containing human beta defensins -3 ( hBD3 ) , connective tissue growth factor gene (CTGF) and enhanced green fluorescent protein (EGFP), and to detect its translation in rabbit bone marrow mesenchymal stem cells (BMSC).Methods The lentivirus containing hBD3, CTGF and EGFP genes was constructed in vitro.The titer of lentivirus was tested with end-paint dilution assay .Rabbit BMSCs were transfected with recombinant virus.The best value of multiplicity of infection (MOI) was tested.The expression condition, transfection efficacy and genetic stability of the target genes were evaluated by using fluorescence microscopy and flow cytometry . Western blotting was used to detect the expression of the target protein .Results Recombinant lentivirus vectors: Lenti-CTGF-hBD3-EGFP, Lenti-hBD3-EGFP, and Lenti-EGFP, were successfully obtained . The titer of the recombinant lentiviruses was 3.21 ×108, 5.80 ×108, and 1.16 ×109, respectively.The best MOI value to transfect BMSCs was 150. The transfection efficacy of these lentivirus vectors was high , reaching 79.72%as assessed by flow cytometry , and it could be stably inherited .Western blotting displayed that target protein expression was successful .Conclusion The construction of recombinant lentiviruses carrying hBD3 and CTGF genes is successful and can be effectively transfected into BMSCs .

Expression and clinical significance of defensin alpha 6 in colorectal cancer

PURPOSE: Defensin alpha 6 (DEFA6) is cationic short peptide with known functional activities in innate antimicrobial immunity. DEFA6 is also highly expressed in colorectal cancer tissue. The mechanism and function of DEFA6 have not been reported how to play a significant role in carcinogenesis and cancer progression. The aim of this study was to evaluate the protumorigenic functions of DEFA6 in the colorectal cancer cell line and the clinical significance of DEFA6 expression in colorectal cancer patients.METHODS: DEFA6 expression was investigated by immunohistochemistry in 151 cases of colorectal cancer tissue and the association of DEFA6 expression was correlated with patient's cancer charecteristics.RESULTS: Immunohistochemistry analysis showed that the DEFA6 protein was expressed higher in advanced cancer group (high T stage, patients with lymph node metastasis, patients with vascular invasion) than early cancer group (low T stage, patients without lymph node metastasis, patients without vascular invasion) (P=0.007/0.032/0.047).CONCLUSION: As patient's survival usually depends on migration and invasion of the cancer cell, the high expression of DEFA6 in colorectal cancer cell is associated with patient's cancer charecteristics and could be a biomarker for colorectal cancer. The future study about characterization of DEFA6 will clarify its specific role in oncogenesis and its therapeutic potential.

β-defensins and the epididymis: Contrasting influences of prenatal, postnatal, and adult scenarios / 亚洲男科学杂志(英文版)

β-defensins are components of host defense, with antimicrobial and pleiotropic immuno-modulatory properties. Research over the last 15 years has demonstrated abundant expression of a variety of β-defensins in the postnatal epididymis of different species. A gradient of region- and cell-specific expression of these proteins is observed in the epithelium of the postnatal epididymis. Their secretion into the luminal fluid and binding to spermatozoa as they travel along the epididymis has suggested their involvement in reproduction-specific tasks. Therefore, continuous attention has been given to various β-defensins for their role in sperm function and fertility. Although β-defensins are largely dependent on androgens, the underlying mechanisms regulating their expression and function in the epididymis are not well understood. Recent investigation has pointed out to a new and interesting scenario where β-defensins emerge with a different expression pattern in the Wolffian duct, the embryonic precursor of the epididymis, as opposed to the adult epididymis, thereby redefining the concept concerning the multifunctional roles of β-defensins in the developing epididymis. In this review, we summarize some current views of β-defensins in the epididymis highlighting our most recent data and speculations on their role in the developing epididymis during the prenatal-to-postnatal transition, bringing attention to the many unanswered questions in this research area that may contribute to a better understanding of epididymal biology and male fertility.

Influence of steroid hormone on expression of Bin1b in primary cultured epididymal epithelial cells / 第二军医大学学报

Objective To study the expression of Bin1b in primary cultured epididymal epithelial cells treated with different steroid hormones at various concentrations. Methods Expression of Bin1b mRNA was examined by agarose gel electrophoresis and PCR in primary cultured epididymal epithelial cells, which were treated with dihydrotestosterone (DHT, 10-9,10-8 and 10-7mol/L), estradiol (E2, 10-9 and 10-8mol/L)) or dexamethasone (Dex, 10-8 and 10-7mol/L) at different concentrations. Results The expression of Bin1b mRNA in primary cultured epididymal epithelial cells was decreased in a time-dependent manner. The mRNA level of Bin1b was decreased by 40% at day 2 and 70% at day 3, and it was hardly detectable at day 5. DHT up-regulated the expression of Bin1b mRNA in a dosage-dependent manner compared with corresponding vehicle control. 10-9mol/L DHT up-regulated Bin1b mRNA level by 37% compared with corresponding vehicle control(P<0.05), and it was further up-regulated after the treatment with 10-8mol/L and 10-7mol/L DHT. E2 and Dex at various concentrations showed no significant effects on the expression of Bin1b mRNA in primary cultured epididymal epithelial cells. Conclusion Dihydrotestosterone can up-regulate the expression of Bin1b mRNA in primary cultured epididymal epithelial cells, while estradio and dexamethasone can not.

Effect of subcutaneous immunotherapy on serum levels of human beta defensin-2 in children with allergic rhinitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the effect of subcutaneousimmunotherapy(SCIT) on levels of the serum human beta defensin-2 in children with allergic rhinitis. METHODS 30 cases of children with allergic rhinitis who were treated by SIT were selected as the treatment group, 20 cases of healthy children as the control group. Serum HBD-2 concentration of the control group was tested. Serum HBD-2 concentration of the treatment group was tested at three different time points: before SCIT, half a year after SCIT and one year after SCIT. And total nasal symptom scores(TNSS) and medication scores were recorded at each time point. RESULTS The serum HBD-2 concentration of the control group, that of the treatment group before SIT, half a year after SIT and one year after SIT were 4.62[4.08; 4.87], 3.74[3.37; 4.61], 4.62[4.13; 5.54], 4.79[4.45;6.19]ng/ml. The HBD-2 concentration gradually increased after SCIT. The TNSS of the treatment group before SCIT, half a year after SCIT and one year after SCIT were 7.43±2.15, 4.17±2.16, 4.20±1.92, The medication scores of the treatment group before SCIT, half a year after SCIT and one year after SCIT were 1.25[0.75; 1.38], 0.25[0; 0.75, 0.25[0; 0.75].There was no correlation (all P>0.05) between the serum HBD-2 concentration and TNSS or medication scores of the treatment group. CONCLUSION The serum levels of HBD-2 in patients with allergic rhinitis were lower than those in normal persons. The specific immunotherapy raised the serum HBD-2 levels of allergic rhinitis patients.