Study on the correlation between telomere dysfunction of human gastric mucosa and chronic atrophic gastritis / 中华消化杂志

Objective:To investigate the correlation between telomere dysfunction of human gastric mucosa and chronic atrophic gastritis (CAG).Methods:From February 12, 2019 to July 10, 2020, at Endoscopy Center, Guang′anmen Hospital, China Academy of Chinese Sciences, 30 patients received endoscopy and pathological diagnosed with CAG (CAG group) were collected, and 30 patients with chronic non-atrophic gastritis (CNAG) were collected at the same time (CNAG group). The relative telomere length was detected by real time fluorescent quantitative polymerase chain reaction. The expression of telomere repeat binding factor (TRF) 1, TRF2 and protection of telomere (POT) 1 at protein level were detected by immunohistochemical staining and semi-quantitative analysis. Spearman analysis was used to analyze the correlation between the relative telomere length of gastric mucosa and the protein expression levels of TRF1, TRF2 and POT1. Mann-Whitney U test and independent sample t test were used for statistical analysis. Results:The relative telomere length of the gastric mucosa in the CAG group was shorter than that in the CNAG group (0.67 (0.51 to 1.17) vs. 1.06(0.69 to 1.37)), and the difference was statistically significant ( U=297.00, P=0.024). The protein expression levels of TRF1, TRF2, and POT1 in the CAG group were all higher than those in the CNAG group, respectively (4.26±2.49 vs. 1.86±1.34, 10.12±2.76 vs. 8.78±2.81, 4.22±2.48 vs. 2.53±1.62), and the differences were statistically significant ( t=8.05, 3.23, 5.39; P<0.001, =0.001, and <0.001). In the CAG group, the protein expression levels of TRF2 and POT1 in gastric mucosa were negatively correlated with the relative telomere length ( r=-0.477 and -0.417, P=0.008 and 0.022). Conclusions:The telomere dysfunction is related to the pathogenesis of CAG. The change of telomere binding protein expression level is involved in the shortening of telomere and pathological process of CAG patients.

Identification of tRNA-Derived Fragments and Their Regulatory Mechanisms on Target Molecules / 中国生物化学与分子生物学报

With the development of sequencing technology and in-depth research on tRNA-derived small molecules (tsRNAs)‚ more and more tRNAs and their functions have been identified in various species. tsRNAs can be divided into tRNA-derived fragment (tRF) and tRNA-derived stress-induced RNA (tiRNA) according to different cleavage sites. And we will focus on tRF‚ which is a kind of non-coding RNA with regulatory function. To deepen the research of tRF‚ a large number of tRF identification methods based on sequencing data and corresponding databases are being constructed in recent years. The former mainly includes the method of Telonis et al. and tDRmapper‚ while the latter mainly includes tRFdb‚ tRF2Cancer and MINTbase. At the same time‚ both provide a more effective tool for the in-depth research of tRF. The regulation mechanisms of tRF are also being illustrated in many studies. tRF mainly regulates the expression of RNA‚ DNA and proteins in a miRNA-like manner. With further investigations‚ researchers have found that tRF also plays a specific regulatory role in various biological processes of human diseases‚ suggesting its role as a potential biomarker. Herein we mainly summarize the identification methods‚ databases‚ regulation mechanisms of tRF and its role in human diseases.

To study the usefulness of CBCL-TRF for assessment and screening of psychiatric morbidity in juvenile delinquent boys in an observation home.

Background: Given the growth of juvenile delinquent population, epidemiologic data on their psychiatric evaluation is becoming increasingly important. Rehabilitation for juvenile delinquent children is the key whether addressing healthcare, poverty, population control, unemployment or human rights issues. Therefore, the present study was undertaken to evaluate the usefulness of CBCL (Child Behaviour Checklist – TRF (Teacher Report Form) for assessment and screening of psychiatric morbidity in juvenile delinquent boys in an observation home. Method: The present cross-sectional study was conducted in an Observation Home for Boys. Prior to conducting the study, informed written permission was sought from the Superintendent of Observation Home for Boys. Study sample consisted of 50 boys aged between 6-16 years. The children were recruited through application of inclusion and exclusion criteria and after taking written informed permission from the Observation Home authorities. Results: Significant CBCL total score was found in 22 (44%) juveniles. Eighteen (36%) juveniles had high score on externalizing behaviour, and 14 (28%) on internalizing behaviour. The sensitivity of CBCL significant score was found to be 88.64% and specificity was 100%. This indicates the utility of CBCL in epidemiological studies and screening of juveniles. It is a simple tool for screening and as noted it has high sensitivity and specificity. Conclusion: In conclusion, the CBCL has implications for the training of manpower for strengthening of mental health services for these children. There is immediate need for multidisciplinary mental health services at each juvenile center.

Time-Restricted Feeding And Brisk Walking in Overweight and Obese Adults

@#Introduction: Obesity and its associated metabolic consequences such as hypertension, type 2 diabetes mellitus and cardiovascular disease are a global epidemic. Conventional treatment of obesity is daily calorie restriction which many patients find challenging. Time-restricted feeding (TRF) is an emerging alternative although very limited scientific evidence is available. Alternatively, brisk walking (BW) has been shown to reduce mortality risks. The objective of this study was to examine the effects of TRF (16 hour/day of fasting) and investigate the additional effects of BW with TRF (16 hour/day of fasting) in overweight and obesity management. Methods: Thirty-six (n=36) overweight and obese participants were enrolled equally into three different groups according to their preferences in this 16- week prospective experimental study; Group A (TRF combined with BW), Group B (TRF alone) and Group C (control group maintaining their lifestyle). Data collection was conducted at the beginning and end of the study for statistical analysis. Results: All Group A and Group B participants showed significant reductions in body mass index, waist circumference, hip circumference, waist-to-hip ratio, body fat percentage, visceral fat level, whole body subcutaneous fat percentage, trunk subcutaneous percentage, legs subcutaneous percentage and arms subcutaneous fat percentage as compared to their control counterparts (Group C) (all p<0.05). However, no significant differences were observed in all anthropometric measurements of Group A participants compared to Group B counterparts. Conclusion: TRF (16 hour/day fasting) can be adopted in weight reduction management of overweight and obese patients. BW for 16 weeks combined with TRF renders no additional effects in overweight and obesity management.

Effects of tRNA and tRNA-derived Fragments on Skeletal Muscle Development / 中国生物化学与分子生物学报

Skeletal muscle is an important tissue of human and livestock. The study of the muscle development is of great significance for treating muscle diseases and improving livestock meat quality. The process of muscle development is controlled by several myogenic transcription factors and signaling pathways. In addition, recent findings established that several noncoding RNAs play a critical role in the regulation of muscle development such as long non-coding RNA (lncRNA), microRNA (miRNA) and circu- lar RNA (circRNA), etc. The detailed mechanism of muscle development is not well understood. Transfer RNAs (tRNAs) are fundamental components in the translation machinery as an adaptor molecule, and tRNA pool could be differentially exploited to modulate expression of mRNAs. In addition, tRNA can be cleaved into tRNA-derived fragments (tRFs) by a variety of ribonucleases (RNases) upon various stress conditions. Unlike the post-transcriptional regulation of lncRNA and miRNA on muscle development, tRNA has been implicated in various aspects of muscle development. Mitochondria play a central role in a plethora of processes related to the maintenance of muscle cellular homeostasis and genomic integrity. Mitochondrial tRNA(mt-tRNA) gene mutations lead to multiple myopathy because human mitochondrial genome is extremely small. The regulation of tRF is similar to miRNAs in regards to the related physiological processes, but are more conservative than miRNA. It is generally believed that tRF has strong tissue specificity, disease specificity and temporal specificity. Some skeletal muscle-specific tRFs could act posttranscriptionally via RNAi or targeting related genes. However, the tRF-sequencing analysis and functional mechanism of tRF are rarely studied in skeletal muscles. The myopathy caused by mitochondrial tRNA gene mutations are particularly complex, which are one of the challenges to diagnose, treat, or prevent diseases. Compared with other noncoding RNAs, the structural complexity of tRF also brings great challenges to data mining and analysis. In this review, we summarize the formation and function of tRNA and tRF especially in muscle development, which will deepen our understandings of related myopathy, and provide new ideas and directions for the investigation of skeletal muscle.

Expression of tRF 31 in breast cancer tissues and its effect on cancer cell proliferation / 医学研究生学报

ObjectiveAt present, there are few reports on the effect of tRF 31 on breast cancer. This study aims to detect and analyze the expression level of tRF 31 in breast cancer tissues and to explore its effect on the proliferation of breast cancer cells.Methods32 tumor tissue samples and the corresponding para-cancer tissues from breast cancer patients in The Affiliated Cancer Hospital of Nanjing Medical University from November 2017 to February 2019 were collected. Six out of the thirty two samples were selected for high-throughput sequencing and at last tRF 31 (FC=6.5781, P=0.023) was selected as the research object. The expression of tRF-31 in cancer and para-cancer tissues was measured by RT-PCR. The sensitivity and specificity of tRF-31 expression in the breast cancer diagnosis were analyzed by ROC curve. Two kinds of human breast cancer cell lines (MDA-MB-231 and MCF-7) were divided into two groups: control group (transfected with negative control) and tRF-31 group (transfected with tRF-31 inhibitor). The proliferation of transfected breast cancer cells was detected by CCK-8 and clonal formation assay. The expression changes of threonine kinase (AKT) and mammalian target of rapamycin (mTOR) in breast cancer cells after transfection were measured to explore the relationship between TRF-31 and AKT/mTOR signaling pathway.ResultsThe expression of tRF-31 in cancer tissues (0.103±0.207) was significantly higher than that in para-cancerous ones (0.028±0.039). ROC curve showed that the sensitivity and specificity of tRF-31 in detecting breast cancer were 90.63% and 53.13%, respectively. In MDA MB 231 cells, the expression of tRF-31 in the tRF-31 group was significantly lower than that in the control group [(0.267±0.012) vs (1±0.040), P<0.01)], and in MCF-7 cells, the expression of tRF-31 in the tRF-31 group was also significantly lower than that in the control group. In MDA-MB-231 and MCF-7 cells, the proliferation ability of the tRF-31 group was lower than that of the control group at 0h, 24h, 48h and 72h. In MDA-MB-231 cells, the clonal formation rate of tRF-31 group [(43.67±3.29)%] was significantly lower than that of the control group [(100±3.74)%] (P<0.01). In MCF-7 cells, the clonal formation rate of tRF-31 group [(49±2.94)%] was significantly lower than that of the control group [(100±4.89)%] (P<0.01). In MDA-MB-231 and MCF-7 cells, the protein levels of AKT and mTOR in tRF-31 group were significantly lower than those in the control group.ConclusiontRF-31 is highly expressed in breast cancer tissues and can promote the proliferation of breast cancer cells. This result is expected to provide a new target for the treatment of breast cancer.

Percepção de professores dos problemas de comportamento em duas amostras clínicas com TDAH: Brasil e Espanha / Teachers' perception of behavioral problems in two clinical samples with ADHD: Brazil and Spain

Considerando a díade professor-aluno no contexto escolar, a figura desse profissional tem sido estimada como uma fonte de informação válida na avaliação comportamental do aluno. Trata-se de uma investigação documental do tipo exploratória-descritiva, retrospectiva e transversal, em que se recorreu a análise de prontuários de crianças e adolescentes entre os anos de 2013 e 2014 diagnosticados com TDAH em duas unidades clínicas de psicologia situadas no Brasil e Espanha. Encontraram-se diferenças significativas apenas em duas síndromes: Problemas de Sociabilidade e Problemas de Atenção. A contribuição deste estudo foi comparar a percepção de professores residentes em diferentes países, e constatar que, independentemente das diferenças culturais, foram encontrados comportamentos similares nos indivíduos com o diagnóstico de TDAH.

Considering teacher-student dyadic interactions, the figure of this professional has been estimated as a valid source of information in the behavioral evaluation of the student. This is an exploratory-descriptive, retrospective and cross-sectional documentary investigation, in which the analysis of medical records of children and adolescents between the years of 2013 and 2014 diagnosed with ADHD in two clinical psychology units located in Brazil and Spain . In the total analysis of TRF scores, significant differences were found only in two syndromes: Sociability Problems and Attention Problems. The contribution of this study was to compare the perception of teachers from different countries and verify that regardless of cultural differences, some behaviors were similarly in individuals with the diagnosis of ADHD.

Research advances of tRNA-derived fragments in tumors / 中国肿瘤临床

The tRNA-derived fragments (tRF and tiRNA) are a newly discovered type of non-coding RNA (ncRNA) that has been found to be stably expressed in peripheral blood. Studies have shown that tRF and tiRNA play important roles in human tumors by regulating multiple processes, including gene expression and silencing, cell proliferation and apoptosis, and protein translation. The tissue-speci-ficity, high abundance, and stability of tRF and tiRNA, along with their broad-spectrum functional roles, confer them significant advan-tages for use in the field of oncology research. There is increasing evidence that aberrantly expressed tRF and tiRNA may be potential biomarkers or therapeutic targets for tumor diagnosis and prognosis. This paper summarizes the source, structure, biological charac-teristics, and functions of different tRF and tiRNA subtypes and explores their potential relationship with tumors and their underlying mechanisms in order to provide a novel idea for the early diagnosis and targeted therapy of tumors.

Development and characterization of polyclonal antibodies against the linker region of the telomere-binding protein TRF2

Background: TRF2 (telomeric repeat binding factor 2) is an essential component of the telomere-binding protein complex shelterin. TRF2 induces the formation of a special structure of telomeric DNA and counteracts activation of DNA damage-response pathways telomeres. TRF2 has a poorly characterized linker region (udTRF2) between its homodimerization and DNA-binding domains. Some lines of evidence have shown that this region could be involved in TRF2 interaction with nuclear lamina. Results: In this study, the fragment of the TERF2 gene encoding udTRF2 domain of telomere-binding protein TRF2 was produced by PCR and cloned into the pET32a vector. The resulting plasmid pET32a-udTRF2 was used for the expression of the recombinant udTRF2 in E. coli RosettaBlue (DE3). The protein was isolated and purified using ammonium sulfate precipitation followed by ion-exchange chromatography. The purified recombinant protein udTRF2 was injected into guinea pigs to generate polyclonal antibodies. The ability of anti-udTRF2 antibodies to bind endogenous TRF2 in human skin fibroblasts was tested by western blotting and immunofluorescent staining. Conclusions: In this study, the recombinant protein udTRF2 and antibodies to it were generated. Both protein and antibodies will provide a useful tool for investigation of the functions of the udTRF2 domain and its role in the interaction between TRF2 and nuclear lamina.

Tocotrienols and its impact in cardiovascular health.

Tocotrienols are members of the vitamin E family essential for human nutrition. It has four isomeric forms i.e., α, β, γ, δ. Palm oil and rice bran oil represent two major nutritional sources of natural tocotrienols. Taken orally, tocotrienols are bioavailable to all vital organs. Apart from tocotrienols antioxidant, anti-inflammatory, antineurodegeneration, antimicrobial, anticancer properties it also has antihypercholesterolemic and antiangiogenic properties. During the last 7 years, tocotrienol research has gained substantial momentum. More than 75% of the entire PubMed literature on tocotrienols has been published on or after 2000. This represents major swing in the overall direction of tocotrienol research. The objective of this review is to highlight the potential significance of the tocotrienol in cardiovascular diseases, mainly anti hypercholesterolemic and anti hyperlipidemic properties along with its efficacy and safety.

A relação entre professores com sofrimento psíquico e crianças escolares com problemas de comportamento / The relation between teachers with psychological distress and schoolchildren with behavior problems

O presente estudo tem por objetivo analisar a relação entre professores com sofrimento psíquico e crianças escolares com problemas de comportamento. Um total de 151 professores avaliou a presença de problemas de comportamento em 372 alunos, através da escala Teacher´s Report Form (TRF). Os alunos foram selecionados através de amostragem por conglomerados e estavam matriculados na primeira série do ensino fundamental. Os professores também responderam a um questionário auto-aplicado a fim de avaliar algumas características pessoais e a presença de sofrimento psíquico (Self Reported Questionnaire SRQ-20). A prevalência de sofrimento psíquico encontrada entre os professores foi de 21,8 por cento. Os resultados mostram percentuais mais elevados na identificação de problemas internalizantes pelas professoras que apresentam sofrimento psíquico.

The present article aims to analyze the relation between teachers suffering from psychological distress and school children with behavioral problems. A total of 372 children were evaluated by teachers through utilizing the Teachers Report Form (TRF). The teachers also answered a questionnaire in order to evaluate some personal characteristics and the presence of psychological distress (Self Reported Questionnaire, SRQ-20). The prevalence of psychological distress found was of 21.8 percent. The results show that the teachers presenting psychological distress identified higher percentages of internalizing problems in the children.

The inhibition of hTERT and TRF2 gene expression and inducing cells apoptosis by adenovirus-mediated hTERT/TRF2 RNA interference in MCF-7 cells / 中国癌症杂志

Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

Research on Fbx4 protein / 国际外科学杂志

Protein degradation mediated by ubiquitin-proteasome pathway is involved in many biological processes.SCF complex is a sort of important E3 ubiquitin-ligase enzyme.Fbx4 protein is one of F-box protein family members,an important component of substrate-specific ScF Fbx4/αβ-crystallin ligase,and a subunit of SCF complex.Inactivation of Fbx4 protein or alteration of its expression level will influence activity of SCF Fbx4/αβ-crystallin ligase,thus will influence expression level of Cyclin D1.And Fbx4 protein is also an important regulatory factor for telomere maintenance.

Design of wireless remote electro-thermometer system / 医疗卫生装备

This paper introduces a wireless remote electro-thermometer system composed of the temperature measurement module, the transmitting module, the receiving module and the computer terminal. The temperature measurement module adopts Texas Instruments' (TI') MSP430 ultra low-power 16-bit microcontroller as the unit for acquisition and control. MSP430 being the controller, the wireless transmitting module and the receiving module are based on TI's TRF6900. The receiving module uses RS232 interface to send the temperature data from the temperature measurement module to the computer terminal.

Application of tailored radiofrequency (TRF) in T2-FLAIR MR imaging / 医疗卫生装备

Objective To investigate the influence of TRF on high-intensity cerebrospinal fluid artifacts in T2-FLAIR MR imaging.Methods In a GE Signa Twinspeed 1.5T system,50 cases were examined with T2-FLAIR sequences including 2 different optional imaging parameters according to the augmentation of TRF.Results TRF pulses significantly improved image quality(P

Gender specific profiles of human telomere shortening during aging / 中国法医学杂志

Objective In human,both in vivo and in vitro,telomere shortening appears to be a major component of cell senescence and aging. However, gender specific human telomere shortening needs to be further characterized. Therefore our study is aimed at clarifying gender-dependent profiles of telomere shortening. Methods 123 peripheral blood samples were collected from healthy individuals of different ages. The mean telomeric restricted fragment (TRF) was measured using Southern Blotting with Dig-labeled probe. Results Distinguished dynamics profiles of telomere shortening were observed among different age groups. Conclusion The result indicates that there are gender specific dynamic profiles of telomere shortening. Therefore, the gender must be considered when an individual age is estimated by telomeric restricted fragment length assay.

Semi - quantitative Comparison of Terminal Restriction Fragment Length and Telomerase in Breast Cancer for Biotherapy / 대한암학회지

PURPOSE: We determined the clinical significance of telomerase activity and telomere length in breast cancer patients and also developed the measuring system of telomerase activity change with RNAse A pre-treatment. MATERIALS AND METHODS: We measured the telomerase activity in 71 breast cancer tissues and paired normal tissues with TRAP (Telomeric Repeat Amplification Protocol) assay. Telomerase activity was calculated by computer-assisted densitometry compared to telomerase activity of the 293 control cell line. To develop the measuring system of telomerase activity modulation, we measured the telomerase activity after the treatment with RNAse A, 150microgram/ml, which inhibited 70% of telomerase activity compared to control in the 293 control cell line. In 59 paired tissues with telomerase activity, terminal restriction fragment (TRFs) length were measured using Southern blotting. RESULTS: Sixty-three out of 71 cancer tissues showed telomerase activity (88.7%), while no telomerase activity was detected in their paired normal tissues. Telomerase activity was correlated to the node metastasis (p=0.02) and stage (p=0.005), but not to the tumor size or the hormonal receptor status. TRFs were neither specific to tumor tissues nor related to any of the clinical parameters. However, changes of TRFs of the tumor tissues from their paired normal tissues were correlated to the telomerase activities. Also the patients with different TRFs between cancer and normal tissues were in more advanced stage. After pre-treatment with the 150microgram/ml of RNAse A, telomerase activity in the tumor tissues showed variable inhibition. Relative inhibition, the ratio of inhibited telomerase activity in each tumor tissue compared to the inhibition of 293 control cell line, was proportional to the telomerase activity. CONCLUSION: In breast cancer, telomerase activity was specific to the tumor tissues and correlated to tumor progression. A combination of telomerase activity and TRFs changes can be used as a guidline in detecting a better candidate for telomerase inhibition. Semi-quantitative assay with RI system can be used in evaluating the changes of telomerase activity after treatment with a new telomerase inhibitor with TRAP assay.