Efecto Antibacteriano del Extracto Etanólico de Equisetum giganteum L. "Cola de Caballo" Frente al Fusobacterium nucleatum, Microorganismo Asociado a la Enfermedad Periodontal

La fitoterapia aplicada a la Odontología se presenta como una eficaz alternativa de tratamiento frente a las enfermedades periodontales (EP) porque busca utilizar los principios activos de las plantas medicinales que se encuentran en gran cantidad en la naturaleza, dándole así las características de ser más asequibles y de menor costo, para combatir los microorganismos patógenos causantes de las EP. El objetivo del estudio fue determinar el efecto antibacteriano in vitro del extracto etanólico de Equisetum giganteum L. frente a cepas ATCC de Fusobacterium nucleatum. El estudio fue de tipo experimental no probabilístico y estuvo constituido en total por 10 placas Petri sembradas con F. nucleatum. Se utilizó extracto etanólico de E. giganteum L. en las concentraciones de 100, 50, 25, 12.5 y 6.25 %. Se utilizó el método de difusión en agar y se incubaron 10 placas a 37 °C durante 07 días. Se midieron los halos de inhibición con un vernier digital, siendo estos datos posteriormente analizados. No se evidenciaron halos de inhibición significativos en ninguno de los discos embebidos con las diferentes concentraciones en las 10 placas Petri sembradas con F. nucleatum, pero sin con la clorhexidina, agente química utilizado como control positivo. En conclusión no se determinó un efecto antibacteriano in vitro del extracto etanólico de E. giganteum L. frente a F. nucleatum, en ninguna de sus concentraciones.

Phytotherapy applied to Dentistry is presented as an effective alternative treatment against periodontal diseases (PD) because it seeks to use the active ingredients of medicinal plants that are found in large quantities in nature, thus giving it the characteristics of being more affordable. and at a lower cost, to combat the pathogenic microorganisms that cause PE. Objective: to determine the in vitro antibacterial effect of the ethanolic extract of Equisetum giganteum L. against ATCC strains of Fusobacterium nucleatum. Material and methods: The study was of a non- probabilistic experimental type and consisted of a total of 10 Petri dishes seeded with F. nucleatum. Ethanolic extract of E. giganteum L. was used in concentrations of 100, 50, 25, 12.5 and 6.25 %. The agar diffusion method was used and 10 plates were incubated at 37 °C for 07 days. The inhibition halos were measured with a digital vernier, and these data were subsequently analyzed. Results: No significant inhibition halos were found in any of the embedded disks with the different concentrations in the 10 Petri dishes seeded with F. nucleatum, but without chlorhexidine, the chemical agent used as a positive control. Conclusions: an in vitro antibacterial effect of the ethanolic extract of E. giganteum L. was not determined against F. nucleatum, in any of its concentrations.

Effects of different debridement methods on titanium surfaces in vitro / 口腔疾病防治

Objective @#To investigate the effect of different decontamination methods, including photodynamic therapy, sandblasting and titanium curette, on titanium surface morphology and bacterial adhesion for the treatment of peri-implant disease. @*Methods@#Porphyromonas gingivalis (Pg) and Fusobacterium nucleatum (Fn) were inoculated on the surface of polished titanium specimens, and titanium specimen surfaces were treated with different decontamination methods after incubation. The titanium specimens were divided into a no-treatment control group, photodynamic group, sandblasting group and titanium curette group according to different decontamination methods. The changes in titanium surface roughness were observed by atomic force microscopy (AFM), and the remaining bacteria on the titanium surface were observed by scanning electron microscopy (SEM) and live/dead bacteria staining tests. After reinoculation of Pg and Fn, bacterial readhesion was observed on the surface of decontaminated titanium specimens. @*Results @#The AFM results showed that the surface roughness of the titanium curette group was significantly higher than that of the no-treatment control group, photodynamic group and sandblasting group (P<0.05), and there was no statistically significant difference between the no-treatment control group, photodynamic group and sandblasting group (P>0.05). The results of contact angle measurement showed that the surface contact angle of each treatment group was smaller than that of the no-treatment control group (P<0.05). The SEM results obtained after the titanium specimen surface was decontaminated showed that the number of bacteria on the no-treatment control group surface was higher and the bacteria were relatively concentrated. The bacteria on the surface of the photodynamic group, sandblasting group and titanium curette group were scattered and distributed in small numbers, and most bacteria on the surface of the photodynamic group were ruptured. The results of the live/dead bacteria staining experiment showed that the percentage of dead bacteria on the surface of the photodynamic group was significantly higher than that of the no-treatment control group, sandblasting group and titanium curette group (P<0.05). The remaining bacteria on the surface of the sandblasting group and titanium curette groups were mainly live bacteria. The remaining bacterial adhesion on the surface was significantly reduced for the sandblasting group compared to the no-treatment control group and the photodynamic and titanium curette groups (P<0.05). SEM and live/dead bacteria staining results of bacterial readhesion on the surface of titanium specimens showed that there was an aggregation of Pg on the surface of the titanium curette group, and its surface bacterial adhesion was significantly higher than that of the no-treatment control group, photodynamic group and sandblasting group. @*Conclusion @#In mechanical decontamination, sandblasting machines are a better option than photodynamic therapy and titanium curettes; however, sandblasting does not remove all bacterial contamination. For sterilization, photodynamic therapy is more effective than sandblasting and titanium curettes. A combination of sandblasting and photodynamic therapy methods for the treatment of peri-implant disease may be considered in clinical practice.

Progress of potential therapeutic targets and small molecule ligands of anti-colorectal cancer pathogenic Fusobacterium nucleatum / 药学学报

The composition of intestinal microflora is closely related to the occurrence and development of colorectal cancer (CRC). Among them, Fusobacterium nucleatum (Fn) has been proved directly related to the recurrence, metastasis and chemotherapy resistance of CRC. Therefore, it is of great significance for the prevention and treatment of colorectal cancer by the exploration potential anti-Fn drug targets and discovery small molecule drugs. However, no selective anti-Fn small molecule inhibitors have been reported so far as well as their anti-Fn thereby "anti-Fn further anticancer" mechanisms are unclear. Herein, this article reviews the potential therapeutic targets and small molecule ligands of Fn in order to provide a reference for the development of anti-Fn and anti-CRC small molecule drugs.

Expression, purification, and characterization of the histidine kinase CarS from Fusobacterium nucleatum / 生物工程学报

Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarScyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarScyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarScyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarScyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.

Clinical Significance of High Expression of Endoplasmic Reticulum Stress-related Proteins Glucose-regulating Protein 78 and X-box Binding Protein 1 induced by Fusobacterium Nucleatum in Esophageal Squamous Cell Carcinoma / 中山大学学报(医学科学版)

ObjectiveTo analyze the induction effect of Fusobacterium nucleatum （Fn） on endoplasmic reticulum stress-related proteins Glucose-regulating protein 78（GRP78） and X-box binding protein 1（XBP1） in esophageal squamous cell carcinoma （ESCC）， and to explore its potential mechanism and clinical significance. MethodsESCC cells KYSE150 and KYSE140 were infected with Fn for 12 h， 24 h and 48 h. The oxidative stress indexes （ROS， MDA and SOD） and the expression of GRP78 and XBP1 in each group were detected by oxidative stress index kit and Western blot. The experiment was divided into Fn groups， Fn+siNC1 groups， Fn+siGRP78 groups， Fn+siNC2 groups and Fn+siXBP1 groups； the oxidative stress indexes， paclitaxel （PTX） response efficacy， abilities of proliferation， invasion and metastasis in each group were compared. The infection of Fn and the expression of GRP78 and XBP1 in 234 ESCC and paracancerous tissues were detected by RNA scope and immunohistochemistry. The correlation between each factor and clinicopathological characteristics of patients was analyzed by Chi-square test. The influence of each factor on the survival of patients was compared by Kaplan-meier survival estimate. ResultsCompared with Fn uninfected KYSE150 and KYSE140 cells， the content of ROS and MDA was gradually increased， the activity of SOD was gradually decreased， and the expression of GRP78 and XBP1 was gradually increased in Fn infected groups （12 h， 24 h and 48 h） （P < 0.05）. Compared with Fn groups， Fn+siNC1 groups， and Fn+siNC2 groups， ROS and MDA contents were decreased， SOD activity was increased， PTX response efficacy was enhanced， and abilities of proliferation， invasion and metastasis were decreased in Fn+siGRP78 and Fn+siXBP1 groups （P < 0.05）. The rates of Fn， GRP78 and XBP1 in ESCC tissues were 43.16%， 69.66% and 60.68%， respectively. And the three indexes were significantly consistent （P < 0.05）. The patients with positive Fn infection and high expression of GRP78 and XBP1 were mostly males with a history of smoking and drinking， and the tumor differentiation degree was low， the invasion degree was deep， the lymph node metastasis rate was high， and the clinical stage was mostly stage Ⅲ/Ⅳ. The 5-year survival time of patients with above positive indexes was shortened （P < 0.05）. ConclusionsFn could induce endoplasmic reticulum stress by inducing the high expression of GRP78 and XBP1， and promote the malignant evolution of ESCC.

Research progress on the relationship between periodontal pathogens and colorectal cancer / 口腔疾病防治

@#With the deepening of the research on the relationship between oral microbiota and systemic diseases, researchers have found that periodontitis is closely related to diabetes, cardiovascular disease, digestive system disease and other systemic diseases. Fusobacterium nucleatum (Fn) and Porphyromonas gingivalis (Pg) are common periodontal pathogens, which play a key role in the occurrence and development of periodontitis. At present, it is also found that Fn and Pg are closely related to the occurrence and development of colorectal cancer (CRC). They can affect the occurrence and development of CRC and the therapeutic effect and prognosis of CRC patients through a variety of ways. It can promote tumor cell proliferation by regulating cell division cycle and inhibiting cell apoptosis, inhibit immune cell function to mediate immune escape and tumor metastasis, and create a pro-inflammatory microenvironment suitable for tumor survival. The study of the effect of periodontal pathogens on the occurrence and development of colorectal cancer and its mechanism also allows us to think about new methods, such as vaccine development, immune agents and antibiotic use to better prevent and treat colorectal cancer and improve the prognosis of patients with colorectal cancer.

Metagenomics analysis reveals universal signatures of the intestinal microbiota in colorectal cancer, regardless of regional differences

The human gut microbiota is a complex and dynamic community of microorganisms living in our intestines and has emerged as an important factor for colorectal adenocarcinoma (CRC). The purpose of our study was to investigate the microbiota composition in Brazilian CRC patients compared with a local control population (CTL) to find out which changes could be considered universal or regional features in CRC microbiota. Fecal samples were obtained from 28 CRC and 23 CTL individuals. The 16S rRNA gene was used for metagenomic analysis. In addition to the anthropometric variables, the clinical stage (TNM 2018) was considered. Patients with CRC had a significant increase in alpha diversity and a higher percentage of genus Prevotella and a decreased proportion of Megamonas and Ruminococcus. Additionally, the proportion of Faecalibacterium prausnitzii was associated with a better prognosis in the first stages of CRC, and Fusobacterium nucleatum proved to be an important marker of colorectal carcinogenesis and tumor aggressiveness. Although regional differences influence the composition of the microbiota, in the case of CRC, the microhabitat created by the tumor seems to be a major factor. Our results contribute to a better understanding of the carcinogenic process, and even in different environments, some factors appear to be characteristic of the microbiota of patients with CRC.

Association between serum IgG titers to periodontal pathogens and outcomes for ischemic stroke patients / 中华老年医学杂志

Objective:To explore the relationship between serum IgG titers to periodontal pathogens and the modified Rankin Score(mRS)after a 3-month follow-up of older patients with ischemic stroke.Methods:Data on 379 older patients diagnosed with ischemic stroke at the Third Affiliated Hospital of Soochow University from January 2019 to April 2020 were retrospectively analyzed.Basic patient data and laboratory test results were collected.Serum samples were collected within three days after the stroke, and serum IgG antibody titers to 9 periodontal pathogens were detected.Patients were followed up for 3 months and the m-RS was calculated.Results:Among the 379 patients, 104(27.4%)had an mRS score ≥ 3 at 3-months.Univariate analysis after adjusting for age and sex showed that atrial fibrillation, ischemic heart disease, ischemic stroke subtypes, the National Institutes of Health Stroke Scale(NIHSS)score, and C-reactive protein(CRP)levels were all risk factors for unfavorable 3-month outcomes(all P<0.05). After fully adjusting confounding factors including age, sex, atrial fibrillation, ischemic heart disease, ischemic stroke subtypes, NIHSS score and CRP level at hospital admission, only the serum IgG antibody titer to Fusobacterium nucleatum increased the risk of unfavorable 3-month outcomes out of titers to 9 periodontal pathogens, and the odds ratio( OR)per standard deviation increase in titer was 3.01(95% CI: 1.73-5.23, P<0.001). Curve fitting showed that the relationship between serum IgG antibody titers to Fusobacterium nucleatum and unfavorable 3-month outcomes was close to a positive linear correlation( χ2=15.333, P<0.001). Stratified analysis showed that there were no subgroup variables, including smoking and drinking habits, comorbidities(hypertension, diabetes, dyslipidemia, chronic kidney disease), and the history of stroke, significantly changed the association between serum IgG antibody titers to Fusobacterium nucleatum and poor prognosis( Pvalues for the interaction were: 0.985, 0.708, 0.388, 0.903, 0.613, 0.700, 0.611). Conclusions:Serum IgG antibody titers to Fusobacterium nucleatum are independently correlated with unfavorable 3-month outcomes in older ischemic stroke patients.The higher the antibody titer, the greater the risk of adverse outcomes.

Research progress of Fusobacterium nucleatum and occurrence and development of colorectal cancer / 国际肿瘤学杂志

Colorectal cancer (CRC) is a common malignant tumor of the digestive tract, which seriously threatens human health. In recent years, many studies have found that Fusobacterium nucleatum (Fn) is positively related to the occurrence of CRC. In the process of CRC carcinogenesis, Fn can play an important role by inducing the expression of pro-inflammatory cytokines and triggering chronic inflammation, inhibiting the function of immune cells, inducing chemotherapy resistance, promoting the expressions of tumor genes and microRNAs and regulating glycolysis.

Advances in the research between Fusobacterium nucleatum and colorectal cancer / 国际外科学杂志

Colorectal cancer is a disease with high morbidity and mortality. Colorectal cancer has a poor prognosis, duing to the current limited treatment methods and can not achieve satisfactory treatment results. Therefore, how to diagnose colorectal cancer patients early and improve the prognosis has always been one of the problems in the medical and scientific research circles. As researchers gradually learn more about the intestinal flora including Fusobacterium nucleatum, the targeted treatment has been applied to the experimental research and clinical treatment of colorectal cancer. This paper reviews the research progress of Fusobacterium nucleatum on the pathogenesis, early diagnosis, prognosis and treatment of colorectal cancer in recent years.

Nivel de higiene y prevalencia de Porphyromona gingivalis y Fusobacterium nucleatum en pacientes recuperados de SARS-CoV-2 / Hygiene level and prevalence of Porphyromona gingivalis and Fusobacterium nucleatum in recovered SARS-CoV-2 patients

Introducción: El SARS-CoV-2 afecta el sistema respiratorio en diferentes grados. La cavidad oral es el lugar más colonizado por bacterias, por lo tanto, al no tener una adecuada higiene pueden presentarse diferentes enfermedades secundarias, lo que ha causado alerta en el gremio odontológico, ya que puede contribuir a complicaciones posteriores en los pacientes. Material y métodos: El estudio fue conformado por 47 pacientes voluntarios recuperados de SARS-CoV-2, residentes de Montemorelos, Nuevo León, México, donde fueron atendidos en Bucalia Dent, consultorio dental. Después del consentimiento informado de cada paciente, se realizó una historia clínica para conocer los síntomas, enfermedades sistémicas, ausencia de dientes y nivel de inflamación gingival de acuerdo al índice de Loe y Silness. A continuación, se tomó una muestra de biofilm microbiano (placa dentobacteriana), la cual se suspendió en una solución buffer de fosfato, posteriormente fue llevada al Centro de Investigación y Desarrollo en Ciencias de la Salud (CIDICS), Monterrey, N.L, México. Se extrajo DNA y se purificó, después se realizó PCR para detectar los patógenos orales; la PCR se visualizó en gel de agarosa (1.5%) por tinción de bromuro de etidio. Resultados: Se detectó 80.85% Porphyromona gingivalis y 68.09% Fusobacterium nucleatum en pacientes recuperados de SARS-CoV-2; 23.4% presentaron inflamación leve de acuerdo al índice de Loe y Silness, 54.5% fueron masculinos y 45.5% femeninos. Por otro lado, 36.4% de los pacientes con inflamación leve tenían de cuatro a seis dientes ausentes. En estos pacientes se detectó 18.18% únicamente con Fusobacterium nucleatum y 27.27% sólo con Porphyromona gingivalis; el sexo masculino tuvo predisposición en 66.6% y el femenino en 33.33%. Se observó infección con los dos patógenos presentes en 45.45%; y 60% de estos pacientes fueron masculinos. Conclusiones: Los pacientes recuperados de SARSCoV- 2 analizados en esta investigación mostraron mala higiene oral y alta prevalencia de los patógenos mencionados altamente relacionados a inflamación gingival o enfermedad periodontal, lo que nos indica que es indispensable la intervención del odontólogo al finalizar el periodo de infección de cada paciente (AU)

Introduction: SARS-CoV-2 affects the respiratory system to different degrees. The oral cavity is a colonized place by bacterias, therefore, by not having good hygiene, different secondary diseases can occur; this has caused an alert in the dental industry, since it can contribute to later complications in patients. Material and methods: The study was conducted in 47 SARS-CoV-2 recovered volunteers from the Montemorelos city of the Nuevo León state, Mexico, who were attended at the Bucalia Dent dental clinic. An informed consent was obtained from each of the patients, then their clinical history was documented in order to know the symptoms, previous systemic diseases, absence of teeth and degree of gingival inflammation, as suggested by Loe and Silness. Subsequently, a dental plaque sample was taken from all patients, which was suspended in a phosphate buffered solution and shipped to The Center for Research and Development in Health Sciences (CIDICS), Monterrey, NL, Mexico for storage. DNA extraction and purification was performed and PCR was carried out for the oral pathogens detection. All PCR products were visualized on 1.5% agarose gel by ethidium bromide staining. Results: Porphyromona gingivalis and Fusobacterium nucleatum were detected in 80.85% and 68.09% of SARS-CoV-2 recovered patients, respectively. 23.4% showed mild inflammation based on the Loe and Silness criteria, 54.5% were male and 45.5% female. On the other hand, 36.4% of patients with mild inflammation had between 4 to 6 missing teeth. A single infection by Fusobacterium nucleatum was detected in 18.18% and by Porphyromona gingivalis in 27.27%; the male sex had a predisposition with 66.66% and 33.33% female; coinfection of both pathogens was observed in 45.45% where 60% were male. Conclusions: SARS-CoV-2 recovered patients show poor oral hygiene and a high prevalence of oral pathogens related to the development of inflammatory gingival or periodontal disease, this suggests the need for an odontological clinical intervention at the end of the course of infection or disease caused by SARS-CoV-2 (AU)

Identification of Proteins Associated with the Formation of Oral Biofilms

ABSTRACT Objective: To identify proteins associated with the formation of Streptococcus gordonii and Fusobacterium nucleatum biofilms. Material and Methods: Biofilms composed of two bacterial species, S. gordonii and F. nucleatum, were cultured for 1, 4, 7, and 10 days. The presence of both species was confirmed via amplification of the srtA and radD genes using real-time PCR. The concentrations of proteins associated with the biofilms and individual species were quantified using Western blotting. Results: The protein profiles of S. gordonii and F. nucleatum from individual cultures determined using one-dimensional electrophoresis revealed proteins found in S. gordonii and in F. nucleatum. Ct and reciprocal Ct values were determined for the exposed S. gordonii and F. nucleatum biofilms. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was detected in biofilms and F. nucleatum, whereas HSP40 protein was present only in biofilms after 7 and 10 days of formation. Conclusion: HSP40 was detected only in the formed biofilms; thus, HSP40 is an essential proteins for adhesion.

Fusobacterium nucleatum promotes TNF-α-induced inflammatory changes in colorectal cancer HCT116 cell through NF-κB signaling pathway / 中华微生物学和免疫学杂志

Objective:To investigate the mechanism by which Fusobacterium nucleatum ( Fn) infection promotes TNF-α-induced inflammatory changes in colorectal cancer HCT116 cells. Methods:Fn-infected cells and TNF-α inflammation induction models were established and divided into 4 groups, namely uninfected control group, Fn-infected group, TNF-α induction group, and Fn+ TNF-α group. First, Fn was used to infect normal colonic epithelial cells hcoEPIC, colorectal cancer HCT116 and LoVo cells, the cell adhesion was detected 4 h later. Subsequently, HCT116 cells were induced with TNF-α for 3 h and then infected with Fn. After 24 h, the cell survival rate and cell damage were detected by CCK8 experiment and lactate dehydrogenase (LDH) viability assay. The ELISA method was further used to detect the expression of nuclear transcription factor NF-κB and cytokines IL-6, IL-8, and IL-1β in the cell and cell culture supernatant. Results:Fn has strong adhesion to colorectal cancer cells HCT116 and LoVo ( P<0.05), but basically does not show invasion. On the contrary, it has a higher invasion rate to normal colonic epithelial cells hcoEPIC after 24 h. Compared with the uninfected Fn group, the cell survival rate of the Fn-infected group was significantly reduced and the cell damage increased ( P<0.001). Three hours after TNF-α induction, Fn infection further promoted cell death and damage ( P<0.001). The expression of NF-κB in the Fn infection and TNF-α alone treatment group was significantly higher than that of the uninfected group ( P<0.001, P<0.05), and the NF-κB expression in the Fn+ TNF-α group was significantly higher than that of the control group and the single treatment group ( P<0.001). In the Fn infection and TNF-α treatment groups, the expressions of IL-6 and IL-8 were significantly higher than those in the uninfected group ( P<0.001), and IL-1β did not change significantly ( P>0.05). The expressions of IL-6, IL-8 and IL-1β in the Fn+ TNF-α group were significantly higher than those in the normal control group and the single treatment group ( P<0.05). Conclusions:Fusobacterium nucleatum can preferentially adhere to colorectal cancer cell HCT116, further promote TNF-α-induced cell damage and death, the expression and release of NF-κB and its downstream pro-inflammatory cytokines IL-6, IL-8, IL-1β.

Research status of fecal detection for colorectal cancer markers / 国际肿瘤学杂志

There are four methods for fecal detection of colorectal cancer (CRC) markers: fecal occult blood test, fecal DNA test, fecal microRNA test, and fecal fusobacterium nucleatum (Fn) test. Fecal immunochemical test has been recommended by experts at home and abroad as the first choice for CRC screening. Fecal DNA test, due to its high price, has not yet been screened for large samples of people in China, so it is recommended as the second level of CRC screening. Fecal microRNA detection has been paid more and more attention by researchers. In recent years, the detection of fecal microbial markers has become more and more popular, especially fecal Fn detection, which is expected to become a microbial indicator for CRC screening.

Research progress on the relationship between Fusobacterium nucleatum and periodontitis / 口腔疾病防治

@#Periodontitis is an infectious disease caused by a variety of microorganisms. Fusobacterium nucleatum is closely related to periodontitis with a high detection rate. Fusobacterium nucleatum is able to coaggregate with other microorganisms and attach and invade epithelial cells with the help of adhesins. It can also promote the occurrence and development of periodontal diseases and even systemic diseases by destroying periodontal tissues with virulence factors and metabolites and inducing a host immune response. However, at present, drugs assisting periodontal nonsurgical treatment clinically cannot target specific periodontal pathogens, such as Fusobacterium nucleatum, which may lead to problems such as dysbacteriosis or drug resistance. Therefore, studies on the pathogenic mechanism of Fusobacterium nucleatum provide new ideas for the prevention and treatment of periodontitis. The idea is to develop materials, drugs, or probiotics that target adhesins, virulence factors, and metabolites or cut off each pathogenic pathway of Fusobacterium nucleatum to inhibit its proliferation and inflammatory responses in deep periodontal pockets and achieve a balance with other oral microorganisms, and the host is beneficial for the control of periodontitis.

Molecular Method of Detection of Primary Endodontic Infection- An Ex Vivo Study.

The removal of microorganism and debris from the root canal system is the prerequisite for success of treatment. This can be achieved by thorough cleaning, shaping and disinfection of the root canal system. The aim of the present study is to investigate the presence of microorganism in primary endodontic infection in South Canara population using PCR technique.METHODSFifty patients with primary endodontic infection were selected for the study. Access cavity preparation was done followed by working length determination and first sample was collected by placing the paper point near the root apex for 1 min and immediately the samples were placed in Tris-EDTA buffer solution, stored at -200 C, followed by PCR analysis of the sample using specific primers for detection of microorganisms.RESULTSA total of 50 cases with primary endodontic infection were analysed for the presence of microorganism within the root canal system. Percentage analysis was done, and the positive results were obtained only for Porphyromonas endodontalis in 50 % of cases.CONCLUSIONSPorphyromonas endodontalis was the prevalent organism seen in primary endodontic infection in this particular geographic distribution.

Correlation of Fecal Fusobacterium nucleatum With Fecal Calprotectin in Patients With Crohn's Disease / 胃肠病学

Background: Fusobacterium nucleatum (Fn) is a common resident of the human GI tract and has been recognized as an opportunistic pathogen implicated in inflammatory bowel disease (IBD). Few studies had focused on the quantitative analysis of Fn in IBD patients. Aims: To establish a novel absolute quantitative real-time PCR method for detection of Fn in fecal samples of Crohn's disease (CD) patients and to study the correlation between fecal Fn and the common inflammatory indicators of CD. Methods: Fecal samples of 57 CD patients and 41 healthy subjects from Suzhou Municipal Hospital were collected and the genomic DNA was extracted. NusG (transcription antitermination protein) of Fusobacterium nucleatum subsp. nucleatum ATCC 25586 was constructed into pUC57 to form a standard plasmid. An absolute quantitative standard curve was built by detecting the gradient diluted standard plasmid via SYBR Green real-time PCR method. The sensitivity, specificity and stability of this novel method were assessed, and then the novel method was used to detect the abundance of Fn in fecal sample of CD patients and healthy controls. Correlations between fecal Fn and fecal calprotectin (FC), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were analyzed. Results: The absolute quantitative real-time PCR method for the detection of fecal Fn established in this study was sensitive, specific and stable. The detection rate and mean abundance of fecal Fn were significantly higher in CD patients than in healthy controls (P=0.034; P=0.039). Fecal Fn abundance in CD patients was positively correlated with FC level (r=0.459, P=0.011). Conclusions: The absolute quantitative real-time PCR established in this study is a promising method for human fecal Fn detection. In Suzhou, Jiangsu Province, the detection rate and abundance of fecal Fn are significantly increased in CD patients. Fecal Fn abundance in CD patients is correlated with FC level, and may reflect the disease activity.

jptsd123 / 口腔疾病防治

@#Esophageal cancer is one of the most lethal digestive system cancers, and its pathogenic factors have always been the focus of research. Recently, it has been found that microorganisms and their metabolites in the esophagus may also represent one of the pathogenic factors. Because of their continuity in anatomical structure, the oral cavity and esophagus have a certain correlation in terms of the composition of flora. In recent years, many scholars have studied the relationship between oral microorganisms and esophageal cancer to monitor changes in oral microorganisms as well as to diagnose and treat esophageal cancer more effectively. In this paper, the research status of oral microorganisms and esophageal cancer was reviewed. The Results of the literature review show that the diversity of bacteria in the esophagus is affected by oral flora in terms of the occurrence and development of esophageal cancer. Among these bacteria, the periodontal red complex, which includes Porphyromonas gingivalis, forsythia and Treponema dentata, as well as common oral microorganisms, such as Streptococcus viridis and Fusobacterium nucleatum, are all related to the occurrence and development of esophageal cancer to a certain extent. At present, there are few studies on the mechanism of microorganisms and esophageal cancer, but scholars have found that lipopolysaccharides and endotoxins, the products of Gram-negative bacteria in the esophagus, may participate in the innate immune response of the host, and the relevant mechanism of action needs further study in order to find new targets for monitoring and treatment.

Study of Fusobacterium nucleatum -related bacterial biofilm promoting M2 polarization of macrophages and chemoresistance in colon cancer / 上海交通大学学报（医学版）

Objective: To investigate whether and how Fusobacterium nucleatum-related bacterial biofilm modulates the infiltration of tumor-associated macrophages into tumor microenvironment and the response to chemotherapy in colon cancer patients. Methods: Both biofilm-based F.n-culture medium (BF-CM) and planktonic F.n-culture medium, (P-CM) Fusobacterium nucleatum was cultured, and the culture-medium was collected to coculture with CRC cell lines and macrophages. Quantified real time PCR( qRT-PCR) was used to measure genes related to chemoresistance, CCK8 assay was conducted to measure proliferation inhibition rate of chemicals to cancer cells, qRT-PCR was used to measure the expression of genes related to macrophage polarization. Fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) were conducted to evaluate existence of bacterial biofilm and infiltration of macrophages in tumor tissues of colon cancer. Results: Expression of chemoresistance-related genes(e.g. MDR1/Abcb1a/ Abcb1b) were higher in BF-CM treated CRC cells than those in P-CM treated cells. CRC cell inhibition level via LOHP/5-FU were reduced with F.n culture medium(metabolites) co-culture, and much lower in BF-CM group. Expression of M2-polarization markers were also higher after BF-CM treated macrophages than P-CM. Biofilm positivity was higher in recurrent(confirmed by PET-CT, CT, or colonoscopy) colon cancer patients with post-resection adjuvant chemotherapy, than that in non-recurrent ones; correlated with infiltration rate of M2 macrophages. Conclusion: Fusobacterium nucleatumrelated bacterial biofilm can induce M2-polarization of intratumor macrophages and could promote chemoresistance to chemicals in CRC cells, which may contribute to prognosis of colon cancer patients.

Comparación del efecto antibacteriano de un extracto etanólico de propóleo a dos concentraciones y del paramonoclorofenol alcanforado frente a Enterococcus faecalis y Fusobacterium nucleatum / Comparison between the antibacterial effects of two concentrations of an ethanolic extract of propolis and camphorated paramonochlorophenol versus Enterococcus Faecalis and Fusobacterium Nucleatum

Objetivo: Comparar el efecto antibacteriano in vitro de un extracto etanólico de propóleo a dos concentraciones frente a Enterococcus faecalis ATCC: 29212 y Fusobacterium nucleatum ATCC: 25586 con el paramonoclorofenol alcanforado (PMCFA). Materiales y método: Se incluyeron dos grupos de 15 placas Petri con cepas activadas de Enterococcus faecalis y Fusobacterium nucleatum. Se elaboró un extracto etanólico a partir de propóleo (EEP), proveniente de la provincia de Oxapampa (Perú), y se diluyó a concentraciones del 20% y el 30%. Se comparó su efecto antibacteriano frente al PMCFA, usando clorhexidina al 2% como control positivo y agua destilada como control negativo; mediante el método de Kirby-Bauer, en un periodo de 7 días para Fusobacterium nucleatum y 24 y 48 horas para Enterococcus faecalis. Se realizó el análisis estadístico mediante el programa SPSS versión 21. Resultados: Frente a Enterococcus faecalis se obtuvieron halos de 10,32 mm, 14,23 mm y 9,10 mm a las 24 horas y halos de 11 mm, 14,96 mm y 8,94 mm a las 48 horas, para las concentraciones de EEP al 20%, el 30% y el PMCFA, respectivamente. Por su parte, frente a Fusobacterium nucleatum, halos de 18,89 mm, 23,17 mm y 13,50 mm para las concentraciones al 20%, el 30% y el PMCFA, respectivamente. Conclusiones: El extracto etanólico elaborado a partir de propóleo de Oxapampa mostró efecto antibacteriano a una concentración del 20% y el 30%, que fue significativamente mayor al del PMCFA, frente a cepas activadas de Enterococcus faecalis y Fusobacterium nucleatum. (AU)

Objective: To compare the antibacterial effect of two concentrations of an ethanolic extract of propolis (EEP) with that of camphorated paramonochlorophenol (CPMC) in Enterococcus faecalis ATCC: 29212 and Fusobacterium nucleatum ATCC: 25586. Material and methods:Two groups of 15 petri plates each, containing activated strains of Enterococcus faecalis and Fusobacterium nucleatum were included in the study. The EEP was prepared using Peruvian propolis from the province of Oxapampa diluted to 20% and 30% of the original concentration. The antibacterial effect of both extracts was compared to CPMC using the Kirby-Bauer disk diffusion method of 07 days for Fusobacterium nucleatum and 24 and 48 hours for Enterococcus faecalis.Statistical analysis was performed using the SPSS program version 21. Results: Inhibitory diameters of 10.32 mm for 20% EEP, 14.23 mm for 30% EEP and 9.10 mm for CPMC were obtained for Enterococcus faecalis at 24 hours, being 11 mm for 20% EEP, 14.96 mm for 30% EEP and 8.94 mm for CPMC at 48 hours. For Fusobacterium nucleatum, the inhibitory diameters were 18.89 mm for 20% EEP, 23.17 mm for 30% EEP and 13.50 mm for CPMC at 7 days. Conclusions: The EEP elaborated from Oxapampa Propolis showed antibacterial effects at concentrations of 20% and 30%, which were significantly higher than those of CPMC in activated strains of Enterococcus faecalis and Fusobacterium nucleatum. (AU)