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Objective To investigate the predictive significance of Septin9 gene methylation(mSEPT9)in peripheral blood in the diagnosis of colorectal adenoma.Methods A total of 52 subjects were included,31 patients with colorectal adenomas were collected as the experimental group in the Department of Pathology and 21 subjects with negative colonoscopy in the gastroenterology outpatient clinic were used as the control group from October 2020 to May 2022.mSEPT9 was detected in the two groups,and the results of CEA level in peripheral blood were collected.All the results were statistically analyzed using the Receiver Operating Characteristic(ROC).Results The area under the curve(area of the ROC:AUC)of mSEPT9detectionto predict the development of adenoma was 0.7205(P<0.05),And the cut-off value(CT value)was 39.55,the corresponding sensitivity was 90.91%and the specificity was 56.67%.The AUC of CEA detection for predicting adenoma was 0.5333(P>0.05).Conclusions The detection of mSEPT9 is better than that of CEA tumor marker detection in peripheral blood for screening colorectal adenomas with good sensitivity and relative specificity.Invasive colonoscopy for people with positive mSEPT9 screening results can be easier to accept by the general population.
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A malária é um problema mundial de saúde, com 249 milhões de casos de infecção, ocasionando 608 mil mortes no ano de 2022. Causada pelo gênero Plasmodium, são cinco principais espécies causadoras da malária no ser humano, o Plasmodium malariae, Plasmodium ovale, Plasmodium knowlesi, Plasmodium vivax e Plasmodium falciparum sendo os dois últimos responsáveis pelo maior número de casos clínicos e mortes ao redor do mundo, transmitida pelo mosquito fêmea do gênero Anopheles durante o repasto sanguíneo. Sabe-se ainda que eritrócitos infectados por Plasmodium berghei ANKA causam alteração no citoesqueleto de actina, consequentemente levando a hiperpermeabiliade da barreira endotelial. Em experimentos in vitro, a imunofluorescência, foi observada alteração do citoesqueleto de actina em células estimuladas com eritrócitos parasitados por PbA (EP), em contrapartida, aquelas não estimuladas (NE) e estimuladas com eritrócitos não parasitados por PbA (EnP), não mostraram alterações no mesmo. Nos experimentos in vivo, ao observar dados coletados, sendo estes respiratórios (penh e frequência respiratória) e parasitemia coletados no 7º DPI, foi observado um mesmo padrão entre o experimento 1 e o experimento 2. Os animais infectados com 106 de eritrócitos infectados, foram alocados em dois grupos, sendo eles hiperparasitemia (HP) ou síndrome do desconforto respiratório agudo-associado a malária (SDRA/SDRA-MA) e comparados àqueles não infectados (NI). Os animais NI, não apresentam parasitemia, em contrapartida, os animais SDRA, tem maior parasitemia que os HP, visto que estes têm aumento em sua parasitemia após o 12º DPI, e assim seguem aumentando gradativamente até levar os animais a óbito.O penh tem o mesmo padrão que a parasitemia, os NI com penh mais baixa que os HPs e os SDRA, sendo dentre estes, o grupo SDRA o mais elevado. A frequência respiratória, por sua vez se apresenta mais elevada no grupo NI, sendo o grupo SDRAmenor que o HP, um achado tido como normal, visto que os pulmões de animais com SDRA sofrem maior dano que os HPs. Apesar de não apresentar um valor significativo, as imagens de gel SDS-PAGE (WB) mostram maior concentração da Septina 9 nos animais com SDRA em comparação com os HPs e com os NIs. O mesmo é observado na qRT-PCR, mesmo sem significância estatística, o valor mostrado nos gráficos temmaior concentração nos SDRA. Assim, a Septina 9 está presente nas CEPP, e, mesmo sem significância estatística, da mesma forma que está presente nas amostras de tecido pulmonar utilizadas no WB e qT-PCR. É hipotetizado ainda que esta proteína pode ser ativada e assim sofrer alteração em sua localização intracelular
Malaria is a global health problem, with 249 million cases of infection, causing 608 thousand deaths in the year 2022. Caused by the genus Plasmodium, there are five main species that cause malaria in humans, Plasmodium malariae, Plasmodium ovale, Plasmodium knowlesi, Plasmodium vivax and Plasmodium falciparum, the last two being responsible for the largest number of clinical cases and deaths around the world, transmitted by the female mosquito of the genus Anopheles during blood meal. It is also known that erythrocytes infected by Plasmodium berghei ANKA (PbA) cause changes in the actin cytoskeleton, consequently leading to hyperpermeability of the endothelial barrier. In in vitro experiments, immunofluorescence, changes in the actin cytoskeleton were observed in cells stimulated with erythrocytes parasitized by PbA (EP), in contrast, those not stimulated (NE) and stimulated with erythrocytes not parasitized by PbA (EnP), did not show changes the same. In the in vivo experiments, when observing collected data, these being respiratory (penh and respiratory frequency) and parasitemia collected on the 7th DPI, the same pattern was observed between experiment 1 and experiment 2. Animals infected with 106 infected erythrocytes were allocated into two groups, namely hyperparasitemia (HP) or malaria-associated acute respiratory distress syndrome (ARDS/ARDS-MA) and compared to those not infected (NI). NI animals do not present parasitemia, on the other hand, ARDS animals have greater parasitemia than HP animals, as the latter have an increase in their parasitemia after the 12th DPI, and thus continue to gradually increase until the animals die. the same pattern as parasitemia, NI with lower penh than HPs and ARDS, among these, the ARDS group being the highest. The respiratory rate, in turn, is higher in the NI group, with the ARDS group being lower than the HP, a finding considered normal, given that the lungs of animals with ARDS suffer greater damage than the HPs. Despite not showing a significant value, SDS-PAGE (WB) gel images show a higher concentration of Septin 9 in animals with ARDS compared to HPs and NIs. The same is observed in qRT-PCR, even without statistical significance, the value shown in the graphs has a higher concentration in ARDS. Thus, Septin 9 is present in CEPP, and, even without statistical significance, in the same way that it is present in lung tissue samples used in WB and qT-PCR. It is also hypothesized that this protein can be activated and thus undergo changes in its intracellular location
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Animaux , Mâle , Souris , Syndrome de détresse respiratoire du nouveau-né/anatomopathologie , Paludisme/anatomopathologie , Contrôle social formel/classification , Techniques in vitro/méthodes , Endothélium , Anopheles/classificationRésumé
OBJECTIVES@#To explore the correlation between cytosine-phosphoric-guanylic (CpG) site of Septin 9 gene and colorectal cancer, and to develop a real-time PCR detection system in plasma in patients with colorectal cancer.@*METHODS@#The methylation of training samples was detected by high-throughput sequencing technology, and the sites highly consistent with the clinical information of colorectal cancer were identified. Then the detection system of real-time PCR was designed to analyze the consistency of plasma and tissue based on methylationa sensitive enzyme digestion. Finally, 100 clinical trials were conducted to evaluate the performance of the detection system with the methylation sensitive enzyme digestion-real-time PCR.@*RESULTS@#The highly consistent sites, which were selected by high-throughput sequencing from 71 training set samples, was the 38th CpG. Based on the detection region, the screened methylation sensitive enzymes were @*CONCLUSIONS@#The 38th CpG site of Septin 9 detected by the detection system of methylation sensitive enzyme digestion-real-time PCR can highly predict the occurrence of colorectal cancer with great clinical application value.
Sujets)
Humains , Tumeurs colorectales/génétique , Ilots CpG/génétique , ADN , Méthylation de l'ADN , Plasma sanguin/métabolisme , Septines/métabolismeRésumé
Objective@#To evaluate the value of methylation detection of plasma Septin9 gene in the diagnosis of colorectal cancer (CRC) and verify its performance. @*Methods@#The plasma samples from 32 CRC patients before colonoscopy and 10 healthy controls during October 2016 and May 2017 were collected, and the methylation levels of Septin9 gene in these samples were detected by the detection kit of plasma Septin9 gene methylation. The coincidence rate, detection limit and precision of the kit in the diagnosis of CRC were evaluated, and its diagnostic value was compared with that of carcinoembryonic antigen (CEA) and facal immunochemical tests (FIT). @*Results@#The positive and negative coincidence rates of the plasma Septin9 gene methylation kit in the detection of CRC were 100%. The reference materials assigned the detection limit were positive, and the coefficient of variation (CV) of precision was less than 5%, which met the basic performance requirements. The sensitivity, specificity, positive predictive value and negative predictive value of the kit in the diagnosis of CRC were 62.50%, 90.00%, 95.20% and 42.90%, respectively. The detection rate of CRC by the kit was 62.50%, significantly higher than those of FIT (28.13%) and CEA (28.13%) (all P<0.05). The area under the ROC curve (AUC ROC ) of the kit in the diagnosis of CRC was 0.762, and the detection rate of stage Ⅰ CRC by the kit was 50.00%. @*Conclusion@#The performance of the plasma Septin9 gene methylation kit meets the anticipated clinical requirements, which may be used as a serological marker for the assistant diagnosis of CRC.
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Objective@#To develop a kind of quality control material which simulates clinical specimens for detecting plasma methylated Septin9 (mSEPT9) and investigate the performance for mSEPT9 detection in external quality assessment (EQA) of laboratories. @*Methods@#The cultured Hela and Jurkat cells, known to contain methylated and unmethylated Septin9 gene respectively, were cultured. The genomic DNA of the cells was collected and extracted, and detected by mSEPT9 kit. According to the Ct value, the genomic DNA was diluted into different concentrations of quality control materials with negative plasma. The homogeneity and stability of the quality control materials were evaluated. The panels consisted of 5 blindly coded samples were distributed to EQA participants and the results were summarized and evaluated. @*Results@#The purity and concentration of extracted genomic DNA met with the needs of use and could be used as quality control products for mSEPT9 detection. The homogeneity and stability met with the requirements of China National Accreditation Service for Conformity Assessment. Some of the participating laboratories occurred false positive results and false negative results, and a good linear correlation for the detected results (Ct values) was only observed in about 55.6% of the laboratories. Among the 9 participating laboratories, 7 laboratories (77.8%) performed well, 1 laboratory (11.1%) qualified, and 1 laboratory (11.1%) unqualified. @*Conclusion@#The quality control materials for mSEPT9 detection was successfully developed and the application in external quality assessment should be of great significance for evaluating and improving the detection ability of clinical laboratory.
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Septin 9 (SEPT9) gene, a member of the conserved framework protein genes family, has guanosine triphosphataseg (GTPase) activity and play an important role in a wide range of biological processes such as cell division, cell polarization and membrane remodeling.Accumulated evidence have confirmed that SEPT9is closely related to the generation and development of human diseases.And SEPT9has become the new aroused general interest in tumor related research at this stage.This article mainly reviews the structural features of SEPT9gene, action mechanism of SEPT9protein and the role of SEPT9gene in a variety of common oncogenesis, and explores the potential value of SEPT9as a marker in early tumor screening.
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Objective To establish the blood Septin9 methylation detection system for early screening of colorectal cancer based on fluorescence PCR technology .Methods The PCR primer of Septin9 was designed by searching the CpG island site of Septin9 methylation in the NCBI database .The high methylation site of Septin9 gene promoter region was confirmed by PCR amplification and sequencing after extracting DNA from colorectal cancer and para-carcinoma tissues .The fluorescence PCR and TaqMan probe detection technique was designed by aiming at high methylation site for constructing the plasma sample methylation detection sys-tem .Then its accuracy ,specificity ,repeatability and minimum detectable amount were performed the assess-ment and analysis .The SETP9 methylation detection was performed in plasma samples from 57 cases of color-ectal cancer and 30 healthy persons .Results The high methylation site of Septin9 gene in tissue samples was confirmed by sequencing .This site served as the target for designing fluorescence PCR detection system .After assessment ,the accuracy ,specificity and repeatability of this detection system were 100% ,the lowest detection amount reached 0 .1 ng/μL .Among the plasma samples in 57 cases of colorectal cancer ,the positive rate of Septin9 methylation site detection was 71 .92% (41/57) and the positive rate of in the patients with pathologi-cal stage Ⅰ and Ⅱ of colorectal cancer reached 64 .2% .But Septin9 gene had no methylation in plasma of healthy population .Conclusion The plasma fluorescence PCR detection system with Septin9 gene methylation as the target has the characteristics of high sensitivity ,high specificity and high accuracy ,which is the reliable detection technique for the early screening of colorectal cancer and has good clinical application prospect .
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Objective:A meta-analysis was use to systematically assess the diagnostic value of Septin-9 in Colorectal cancer. Methods:Literature fulfilling the criteria was searched in PubMed, Foreign Medical Journals Platform, Ovid, CNKI, CBM, WanFang and VIP Databases from inception to Jan. 2017. Literatures were strictly screened according to the inclusion and exclusion cri-teria. Study quality was assessed in terms of the Quality Assessment of Diagnostic Accuracy Studies ( QUADAS) checklist. A bivariate Meta-analysis model was employed to assess the pooled accuracy,and study heterogeneity was evaluated via Cochran-Q and I2 tests;subgroup analysis and sensitivity analysis were conducted to deeply trace the sources of heterogeneity;publication bias was judged by Deek′s funnel plot. Results:A total of 11 studies were included. Analysis of methylated Septin-9 achieved a pooled sensitivity of 0. 70 (95%CI:0. 67-0. 72),specificity of 0. 91 (95%CI:0. 90-0. 92),and DOR of 28. 76(95%CI:17. 70-46. 75),corresponding to an AUC of 0. 9221. Heterogeneity test suggested that there was obvious heterogeneity from non-threshold effect. Sensitivity analysis identified one outlier study. Subgroup analysis results showed that the AUC of 1/3 positive to 2/3 positive group was 0. 9397 versus 0. 8265,and the AUC of the Asian population group to the Caucasian population group was 0. 9368 versus 0. 9210. Funnel plot ( Deek′s) revealed no publication bias. Conclusion:Our data indicate that circulating methylated Septin-9 seemed to harbor a relatively high accuracy in conforming colorectal cancer,and might be popularized as a routine biomarker for colorectal cancer detection.
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Objective:A meta-analysis was use to systematically assess the diagnostic value of Septin-9 in Colorectal cancer. Methods:Literature fulfilling the criteria was searched in PubMed, Foreign Medical Journals Platform, Ovid, CNKI, CBM, WanFang and VIP Databases from inception to Jan. 2017. Literatures were strictly screened according to the inclusion and exclusion cri-teria. Study quality was assessed in terms of the Quality Assessment of Diagnostic Accuracy Studies ( QUADAS) checklist. A bivariate Meta-analysis model was employed to assess the pooled accuracy,and study heterogeneity was evaluated via Cochran-Q and I2 tests;subgroup analysis and sensitivity analysis were conducted to deeply trace the sources of heterogeneity;publication bias was judged by Deek′s funnel plot. Results:A total of 11 studies were included. Analysis of methylated Septin-9 achieved a pooled sensitivity of 0. 70 (95%CI:0. 67-0. 72),specificity of 0. 91 (95%CI:0. 90-0. 92),and DOR of 28. 76(95%CI:17. 70-46. 75),corresponding to an AUC of 0. 9221. Heterogeneity test suggested that there was obvious heterogeneity from non-threshold effect. Sensitivity analysis identified one outlier study. Subgroup analysis results showed that the AUC of 1/3 positive to 2/3 positive group was 0. 9397 versus 0. 8265,and the AUC of the Asian population group to the Caucasian population group was 0. 9368 versus 0. 9210. Funnel plot ( Deek′s) revealed no publication bias. Conclusion:Our data indicate that circulating methylated Septin-9 seemed to harbor a relatively high accuracy in conforming colorectal cancer,and might be popularized as a routine biomarker for colorectal cancer detection.