ABSTRACT
Resumo Objetivo Desenvolver uma modalidade de educação em saúde baseado na teoria do comportamento planejado (TCP) e explorar sua influência no estilo de vida pós-operatório de pacientes com câncer cervical. Métodos Um total de 122 pacientes diagnosticados de janeiro de 2019 a junho de 2020 foram divididos nos grupos Controle e Educação (n = 61) usando uma tabela de números aleatórios. O grupo Controle recebeu educação em saúde tradicional após a cirurgia, enquanto o grupo Educação recebeu educação em saúde baseada em TCP após a cirurgia e seus dados gerais, adesão à medicação, adesão à dieta, tempo de alta, incidência de complicações, estilo de vida, qualidade de vida (QV) e estado mental antes e depois da cirurgia foram comparados. Resultados Não foram encontradas diferenças significativas nos dados gerais, pontuação de estilo de vida pré-operatório, pontuação de qualidade de vida e pontuação de estado mental entre os dois grupos (P>0,05). No grupo Educação, as pontuações de adesão à medicação e adesão à dieta foram significativamente maiores do que no grupo Controle, enquanto o tempo de alta e a incidência total de complicações foram mais precoces e menores do que no grupo Controle (P <0,05). Após a cirurgia, as pontuações de autorrealização, relacionamento interpessoal, nutrição, atividade física, responsabilidade pela saúde, gestão do estresse, dimensão física, dimensão social/familiar, dimensão emocional e dimensão funcional aumentaram significativamente, enquanto as pontuações do termômetro de sofrimento e da lista de problemas diminuíram em ambos os grupos, e a melhora nas pontuações foi maior no grupo Educação do que o grupo Controle (P <0,05). Conclusão A modalidade de educação em saúde baseada na TCP pode melhorar a adesão ao tratamento pós-operatório, reduzir as complicações pós-operatórias e melhorar o estilo de vida, a qualidade de vida e o estado mental de pacientes com câncer cervical.
Resumen Objetivo Elaborar una modalidad de educación para la salud basada en la teoría del comportamiento planeado (TCP) e investigar su influencia en el estilo de vida posoperatorio de pacientes con cáncer cervical. Métodos Un total de 122 pacientes diagnosticados de enero de 2019 a junio de 2020 fueron divididos entre los grupos Control y Educación (n=61) por medio de una planilla de número aleatorios. El grupo Control recibió educación para la salud tradicional luego de la cirugía, mientras que el grupo Educación recibió educación para la salud basada en la TCP después de la cirugía; se compararon los datos generales, la adherencia a la medicación, la adherencia a la dieta, el tiempo de alta, la incidencia de complicaciones, el estilo de vida, la calidad de vida (CV) y el estado mental antes y después de la cirugía. Resultados No se encontraron diferencias significativas entre los dos grupos en datos generales, puntaje de estilo de vida preoperatorio, puntaje de calidad de vida y puntaje de estado mental (P>0,05). En el grupo Educación, los puntajes de adherencia a la medicación y adherencia a la dieta fueron significativamente más altos que en el grupo Control; mientras que el tiempo de alta fue anterior y la incidencia total de complicaciones fue menor que en el grupo Control (P <0,05). Después de la cirugía, los puntajes de autorrealización, relaciones interpersonales, nutrición, actividad física, responsabilidad por la salud, gestión del estrés, dimensión física, dimensión social/familiar, dimensión emocional y dimensión funcional aumentaron significativamente; mientras que los puntajes del termómetro de sufrimiento y de la lista de problemas disminuyeron en ambos grupos y la mejora de los puntajes fue mayor en el grupo Educación que en el grupo Control (P <0,05). Conclusión La modalidad de educación para la salud basada en la TCP puede mejorar la adherencia al tratamiento posoperatorio, reducir las complicaciones posoperatorias y mejorar el estilo de vida, la calidad de vida y el estado mental de paciente con cáncer cervical.
Abstract Objective To develop a health education mode based on the theory of planned behavior (TPB), and to explore the influence of this mode on the postoperative lifestyle of patients with cervical cancer. Methods A total of 122 patients diagnosed from January 2019 to June 2020 were divided into control and education groups (n=61) using a random number table. Control group received traditional health education after operation, while education group was given TPB-based health education after operation. Their general data, medication compliance, dietary compliance, discharge time, incidence of complications, lifestyle, quality of life (QOL) and mental state before and after operation were compared. Results No significant differences were found in general data, preoperative lifestyle score, QOL score and mental state score between the two groups (P>0.05). In education group, the medication compliance and dietary compliance scores were significantly higher than those in control group, while the discharge time and the total incidence of complications were earlier and lower than those in control group (P<0.05). After operation, the scores of self-realization, interpersonal relationship, nutrition, physical activity, health responsibility, stress management, physical dimension, social/family dimension, emotional dimension and functional dimension significantly rose, while the distress thermometer and problem list scores declined in both groups, and education group had better improvement in scores than control group (P<0.05). Conclusion Theory of planned behavior health education mode can improve the postoperative treatment compliance, reduce postoperative complications, and ameliorate the lifestyle, quality of life and mental state of patients with cervical cancer.
ABSTRACT
Objective:To investigate the rehabilitation resources of community health centers in Shanghai. Methods:December, 2018, all 247 community health centers in 16 districts of Shanghai were investigated with Health Institution Questionnaires, including the number of rehabilitation personnel (physiatrician, physical therapists) and the number of rehabilitation beds, etc. Results:There were 152 physiatrician and 597 physical therapists in community health centers in Shanghai. The numbers of physiatrician and physical therapists per 1000 registered population were 0.01 and 0.04, and were 0.03 and 0.12 for per 1000 registered elderly population. There were 17 484 beds in community health centers, in which the rehabilitation beds were 1425, accounting for 8.15%. Besides, the numbers of rehabilitation beds per 1000 registered population and registered elderly population in Shanghai were 0.10 and 0.30. Conclusion:The development of community-based rehabilitation in Shanghai is unbalanced. There is lack of community-based rehabilitation resources, the supply of service is insufficient, and there is gap between supply and demand. It is suggested to strengthen the support to community-based rehabilitation, and improve the supply of community-based rehabilitation resources.
ABSTRACT
With the development of big data, the Internet and information technology, artificial intelligence is gradually being applied to the field of hospital management. The authors used advanced artificial intelligence technology to conduct structured collection and monitor the hospital′s current medical quality data, identifying key points and key events affecting medical quality and safety from the link, and providing medical personnel′s behavior with pre-warning, process monitoring and post-evaluation to improve hospital medical quality management.
ABSTRACT
With the development of big data, the Internet and information technology, artificial intelligence is gradually being applied to the field of hospital management. The authors used advanced artificial intelligence technology to conduct structured collection and monitor the hospital′s current medical quality data, identifying key points and key events affecting medical quality and safety from the link, and providing medical personnel′s behavior with pre-warning, process monitoring and post-evaluation to improve hospital medical quality management.
ABSTRACT
The association between serum uric acid and the risk of incident diabetes in Chinese adults remains unknown. This study aimed to investigate this association in a community-dwelling population aged ≥ 40 years in Shanghai, China. Oral glucose tole3rance test was conducted during baseline and follow-up visits. Relative risk regression was utilized to examine the associations between baseline gender-specific serum uric acid levels and incident diabetes risk. A total of 613 (10.3%) incident diabetes cases were identified during the follow-up visit after 4.5 years. Fasting plasma glucose, postload glucose, and glycated hemoglobin A1c during the follow-up visit progressively increased across the sex-specific quartiles of serum uric acid (all Ps < 0.05). The incidence rate of diabetes increased across the quartiles of serum uric acid (7.43%, 8.77%, 11.47%, and 13.43%). Multivariate adjusted regression analysis revealed that individuals in the highest quartile had 1.36-fold increased risk of diabetes compared with those in the lowest quartile of serum uric acid (odds ratio (95% confidence interval) = 1.36 (1.06-1.73)). Stratified analysis indicated that the association was only observed in women. Accordingly, serum uric acid was associated with the increased risk of incident diabetes among middle-aged and elderly Chinese women.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , China/epidemiology , Diabetes Mellitus/epidemiology , Incidence , Prospective Studies , Risk Factors , Uric AcidABSTRACT
Objective To investigate the association between levels of serum alanine aminotransferase (ALT) and the risks of cardiovascular disease in Chinese. Methods A survey of 10 375 subjects aged 40 or older in Jiading District of Shanghai was performed using cluster sampling. All the subjects were recruited to undergo questionnaire interview, anthropometric measurements, and fasting blood sampling. A total of 8 877 participants were included in the final analyses, including 5 664 females and 3 213 males. The mean age of individuals was ( 58.68 ± 9.22)yearsinmenand(57.65±8.93)yearsinwomen(P＜0.01).10-yearriskforafirsthardatherosclerotic cardiovascular diseases (ASCVD) event was calculated. Results Increased ALT levels were associated with more unfavorable cardiovascular risk profiles and 10-year risk for ASCVD. Increased ALT levels were associated with the prevalence of diabetes mellitus in female participants, while in male participants the prevalence of diabetes mellitus appeared as a U-distribution. The prevalence of hypertension and hyperlipidemia were 66.04%, 52.74%and 64.49%, 47.63% respectively. Increased serum ALT was associated with higher prevalence of 10-year risk for ASCVD. As compared with participants in the first ALT quartile, the risks of 10-year risk for ASCVD were increased by 148%(OR＝2.48, 95%CI 1.78-3.45) and 139%(OR＝2.39, 95%CI 1.78-3.19), respectively in male and female participants. As compared with participants in the first ALT quartile, higher prevalence of 10-year risk for ASCVD were increased by 96%(OR＝1.96, 95%CI 1.30-2.95), while there was no significantly statistical difference in men. Conclusion A high-normal serum ALT level was significantly associated with an increased risk of the cardiovascular disease in middle-aged and elderly Chinese.
ABSTRACT
OBJECTIVE@#To examine the association between serum uric acid levels and cardiovascular disease risk among individuals without diabetes.@*METHODS@#We investigated the association between serum uric acid levels and the risk of prevalent cardiometabolic diseases, 10-year Framingham risk for coronary heart disease, and 10-year risk for atherosclerotic cardiovascular diseases (ASCVD) among 8,252 participants aged ⪖ 40 years without diabetes from Jiading district, Shanghai, China.@*RESULTS@#Body mass index, waist circumference, blood glucose, glycated hemoglobin, blood pressure, and serum lipids increased progressively across the sex-specific quartiles of uric acid (all P trend < 0.05). Compared with individuals in the lowest quartile, those in the higher quartiles had a significantly higher prevalence of obesity, hypertension, and dyslipidemia (all P trend < 0.05). A fully adjusted logistic regression analysis revealed that individuals in the highest quartile had an increased risk of predicted cardiovascular disease compared with those in the lowest quartile of uric acid. The multivariate adjusted odds ratios (ORs) [95% confidence intervals (CIs)] for the highest quartiles for high Framingham risk were 3.00 (2.00-4.50) in men and 2.95 (1.08-8.43) in women. The multivariate adjusted ORs (95% CIs) for the highest quartile for high ASCVD risk were 1.93 (1.17-3.17) in men and 4.53 (2.57-7.98) in women.@*CONCLUSION@#Serum uric acid level is associated with an increased risk of prevalent obesity, hypertension, dyslipidemia, 10-year Framingham risk for coronary heart disease, and 10-year risk for ASCVD among Chinese adults without diabetes.
Subject(s)
Female , Humans , Male , Middle Aged , Biomarkers , Blood , Blood Glucose , Blood Pressure , Body Mass Index , China , Coronary Disease , Blood , Epidemiology , Cross-Sectional Studies , Glycated Hemoglobin , Lipids , Blood , Predictive Value of Tests , Prevalence , Risk Factors , Sex Factors , Uric Acid , BloodABSTRACT
Objective To explore the effect of Balint group training on the humanistic caring ability of junior nurses. Methods About 96 junior nurses from the department of oncology were randomly assigned to an intervention group and a control group equally.Balint group training was given to the intervention group every 2 weeks in a year.The control group completed the humanistic care training according to the regular procedure in the department of the hospital.At the beginning of the study and at the end of the study,the nursing staff from the two groups and the patients under their continuous nursing care for more than 3 days were surveyed by the care efficiency scale,nursing care behavior scale and patient care perception questionnaire.Results Before the intervention,there was no significant difference between the groups in the care performance and behavior of nursing staff and the patients'care perception (P>0.05).After the intervention there was significant difference in the care performance and behavior of nursing staff and the patients' care perception (P<0.05). Conclusions Balint group training can improve the caring performance of junior nurses.Their ability to express care and establish the caring nurse-patient relationship can be improved by this group training.They become more voluntary to integrate caring behavior into the daily care and their caring behaviors can be more likely to be felt and recognize by the patients.
ABSTRACT
Objective·To investigate the relationship between obesity and chronic kidney disease (CKD) in a community-based study among a middle-aged and elderly Chinese population. Methods·A survey of 10375 inhabitants aged 40 years or older in Jiading District of Shanghai was performed using cluster sampling. All the subjects were recruited to undergo questionnaire interview, anthropometric measurements, and biochemical measurements. Blood and urine samples were collected to measure biochemical features, including plasma glucose, lipid profiles, liver and renal function, as well as urinary albumin and creatinine. Obesity status was diagnosed according to the World Health Organization (WHO) criteria. According to body mass index (BMI) levels, the subjects were divided into normal weight group (BMI<25 kg/m2), overweight group (25 kg/m2 ≤ BMI<30 kg/m2) and obesity group (BMI ≥ 30 kg/m2). The estimated glomerular filtration rate (eGFR) was calculated based on American Chronic Kidney Disease-Epidemiology Collaboration (CKD-EPI) equation. Logistic regression analysis was used to analyze the risk of CKD among people with different BMI levels. Results·Increased BMI levels were associated with more unfavorable metabolic profiles. After a variety of factors were adjusted,each 1 kg/m2 increment of BMI was associated with a 9% higher risk of prevalent CKD, and the odds ratio (OR) was 1.09 (95% CI:1.07-1.12). Results from multivariate Logistic regression analysis showed that in the total population, compared with individuals in the normal weight group, those in the overweight and obesity groups had 51% (OR=1.51, 95% CI: 1.27-1.78) and 181% (OR=2.81, 95% CI: 2.20-3.61) higher risk of prevalent CKD. Conclusion·In the middle-aged and elderly community population in China, overweight and obesity are associated with an increased risk of prevalent CKD.
ABSTRACT
Objective·To investigate the relationship between obesity and chronic kidney disease (CKD) in a community-based study among a middle-aged and elderly Chinese population. Methods·A survey of 10375 inhabitants aged 40 years or older in Jiading District of Shanghai was performed using cluster sampling. All the subjects were recruited to undergo questionnaire interview, anthropometric measurements, and biochemical measurements. Blood and urine samples were collected to measure biochemical features, including plasma glucose, lipid profiles, liver and renal function, as well as urinary albumin and creatinine. Obesity status was diagnosed according to the World Health Organization (WHO) criteria. According to body mass index (BMI) levels, the subjects were divided into normal weight group (BMI<25 kg/m2), overweight group (25 kg/m2 ≤ BMI<30 kg/m2) and obesity group (BMI ≥ 30 kg/m2). The estimated glomerular filtration rate (eGFR) was calculated based on American Chronic Kidney Disease-Epidemiology Collaboration (CKD-EPI) equation. Logistic regression analysis was used to analyze the risk of CKD among people with different BMI levels. Results·Increased BMI levels were associated with more unfavorable metabolic profiles. After a variety of factors were adjusted,each 1 kg/m2 increment of BMI was associated with a 9% higher risk of prevalent CKD, and the odds ratio (OR) was 1.09 (95% CI:1.07-1.12). Results from multivariate Logistic regression analysis showed that in the total population, compared with individuals in the normal weight group, those in the overweight and obesity groups had 51% (OR=1.51, 95% CI: 1.27-1.78) and 181% (OR=2.81, 95% CI: 2.20-3.61) higher risk of prevalent CKD. Conclusion·In the middle-aged and elderly community population in China, overweight and obesity are associated with an increased risk of prevalent CKD.
ABSTRACT
Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Subject(s)
Aminoacyltransferases , Metabolism , Bacterial Proteins , Metabolism , Cyclization , Cysteine Endopeptidases , Metabolism , Enzymes, Immobilized , Metabolism , Kinetics , Peptides , Metabolism , Peptides, Cyclic , Staphylococcus aureusABSTRACT
Three cyclotides were isolated from the whole plant of Viola yedoensis in this study. The two, vary peptide E and cycloviolacin Y5, were previously reported, and a novel cycloviolacin VY1 was characterized according to the interpretation of MS/MS fragmentation of peptides which were produced from the reduced and alkylated parent peptide with the digestion of Endo Lys-C, trypsin and chymotrypsin, separately. The stability of remarkable resistance to proteolytic degradation by trypsin and chymotrypsin, and that of thermal denaturation was confirmed again. Besides, the IC50 value of cycloviolacin VY1 against influenza A H1N1 virus was (2.27 +/- 0.20) microg x mL(-1). It is the first cyclotide reported with anti-influenza A H1N1 virus activity in vitro assay.
Subject(s)
Antiviral Agents , Pharmacology , Cyclotides , Pharmacology , Influenza A Virus, H1N1 Subtype , Tandem Mass Spectrometry , Viola , ChemistryABSTRACT
The synthetic biology matures to promote the heterologous biosynthesis of the well-known drug paclitaxel that is one of the most important and active chemotherapeutic agents for the first-line clinical treatment of cancer. This review focuses on the construction and regulation of the biosynthetic pathway of paclitaxel intermediates in both Escherichia coli and Saccharomyces cerevisiae. In particular, the review also features the early efforts to design and overproduce taxadiene and the bottleneck of scale fermentation for producing the intermediates.
Subject(s)
Alkenes , Chemistry , Metabolism , Antineoplastic Agents, Phytogenic , Chemistry , Metabolism , Biosynthetic Pathways , Diterpenes , Chemistry , Metabolism , Escherichia coli , Metabolism , Fermentation , Metabolic Engineering , Paclitaxel , Chemistry , Metabolism , Prodrugs , Saccharomyces cerevisiae , Metabolism , Synthetic BiologyABSTRACT
Abstract: The first-line drug artemisinin is widely used against malaria. Commercially available artemisinin is extracted from plants. However, the lack of sufficient raw material, artemisinin and the cost associated with the drug's manufacture have limited the supply of ACT to most malaria sufferers in the Developing World. As such, it is important to develop a low cost, fine to environment and high-quality method to supply sufficient and reliable quantities of artemisinin in the future. The field of synthetic biology, which utilizes cell factories to manipulate microbial metabolism to enhance the production of artemisinin and its intermediates, has a particularly strong impact by providing new platforms for chemical production. After a brief introduction of the artemisinin biosynthetic pathway, the present review focuses on the introduction of artemisinin biosynthetic genes, such as the genes encoding amorpha-4, 11-diene monooxygenase, NADPH: cytochrome P450 oxidoreductase, artemisinic aldehyde delta 11(13) reductase and aldehyde dehydrogenase. The review also addresses general considerations for potential contributions of synthetic biology to artemisinin production, with an emphasis on factors influencing interest compounds production in chassis cells.
Subject(s)
Antimalarials , Metabolism , Artemisinins , Metabolism , Biosynthetic Pathways , Cytochrome P-450 Enzyme System , Genetics , Escherichia coli , Metabolism , Gene Dosage , Genetic Engineering , Isoenzymes , Genetics , RNA Nucleotidyltransferases , Genetics , Retinal Dehydrogenase , Genetics , Saccharomyces cerevisiae , Metabolism , Synthetic BiologyABSTRACT
Synthetic biology of natural products is the design and construction of new biological systems by transferring a metabolic pathway of interest products into a chassis. Large-scale production of natural products is achieved by coordinate expression of multiple genes involved in genetic pathway of desired products. Promoters are cis-elements and play important roles in the balance of the metabolic pathways controlled by multiple genes by regulating gene expression. A detection plasmid of Saccharomyces cerevisiae was constructed based on DsRed-Monomer gene encoding for a red fluorescent protein. This plasmid was used for screening the efficient promoters applying for multiple gene-controlled pathways. First of all, eight pairs of primers specific to DsRed-Monomer gene were synthesized. The rapid cloning of DsRed-Monomer gene was performed based on step-by-step extension of a short region of the gene through a series of PCR reactions. All cloned sequences were confirmed by DNA sequencing. A vector named pEASYDs-M containing full-length DsRed-Monomer gene was constructed and was used as the template for the construction of S. cerevisiae expression vector named for pYeDP60-Ds-M. pYeDP60-Ds-M was then transformed into S. cerevisiae for heterologous expression of DsRed-Monomer gene. SDS-PAGE, Western blot and fluorescence microscopy results showed that the recombinant DsRed-Monomer protein was expressed successfully in S. cerevisiae. The well-characterized DsRed-Monomer gene was then cloned into a yeast expression vector pGBT9 to obtain a promoter detection plasmid pGBT9Red. For determination efficacy of pGBT9Red, six promoters (including four inducible promoters and two constitutive promoters) were cloned by PCR from the S. cerevisiae genome, and cloned into pGBT9Red by placing upstream of DsRed-Monomer gene, separately. The fluorescence microscopy results indicated that the six promoters (GAL1, GAL2, GAL7, GAL10, TEF2 and PGK1) can regulate the expression of DsRed-Monomer gene. The successful construction of pGBT9Red lays the foundation for further analysis of promoter activity and screening of promoter element libraries.
Subject(s)
Amino Acid Sequence , Base Sequence , Genetics , Cloning, Molecular , DNA Primers , Gene Expression Regulation, Fungal , Genetic Vectors , Luminescent Proteins , Genetics , Metabolism , Plasmids , Genetics , Promoter Regions, Genetic , Genetics , Recombinant Proteins , Genetics , Saccharomyces cerevisiae , Genetics , Metabolism , Synthetic Biology , Transformation, GeneticABSTRACT
<p><b>OBJECTIVE</b>To explore whether hepatitis C virus core protein (HCV C) regulates the expression of NFAT1 to participate in the progression and malignant biological behavior of intrahepatic cholangiocarcinoma cells.</p><p><b>METHODS</b>The recombinant plasmid pEGFP-N(3)-HCV C and the empty vector pEGFP-N(3) were cotransfected with enhanced green fluorescent protein (EGFP) into RBE cells using liposome. Real-time PCR and Western blotting were used to examine the expression of NFAT1 mRNA and protein in the transfected RBE cells. MTT assay was used to evaluate the changes in the cell proliferation, and the cell cycle changes were analyzed by flow cytometry.</p><p><b>RESULTS</b>HCV C transfection significantly enhanced the expressions of NFAT1 mRNA and protein in RBE cells (P<0.05) and promoted the progression of cell cycle into G(2)/M phase to accelerate the cell proliferation.</p><p><b>CONCLUSION</b>Transfection with HCV C gene up-regulates NFAT1 expression and promotes the cell cycle progression and proliferation of intrahepatic cholangiocarcinoma cells, suggesting the involvement of HCV C in the progression of intrahepatic cholangiocarcinoma.</p>
Subject(s)
Humans , Bile Duct Neoplasms , Pathology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cholangiocarcinoma , Pathology , Gene Expression , NFATC Transcription Factors , Genetics , Plasmids , Transfection , Viral Core Proteins , GeneticsABSTRACT
Human enterovirus 71 (EV71) is one of the major etiological agents for the hand, foot, and month disease (HFMD) and is causing frequent, widespread occurrence in the mainland of China. The single positive-stranded RNA genome of EV71 is translated into a single polyprotein which is autocleavaged into structural and nonstructural proteins. The functions of many nonstructural proteins characterized in the life cycle of virus are potential targets for blocking viral replication. This article reviews the studies of the structures and functions of nonstructural proteins of EV71 and the anti-enterovirus 71 drugs targeting on these nonstructural proteins.
Subject(s)
Humans , Antiviral Agents , Pharmacology , Enterovirus A, Human , Genetics , Hand, Foot and Mouth Disease , Drug Therapy , Virology , Molecular Targeted Therapy , Peptide Hydrolases , Chemistry , Metabolism , Physiology , Protein Kinase Inhibitors , Pharmacology , RNA, Viral , Genetics , Viral Nonstructural Proteins , Chemistry , Metabolism , Physiology , Virus ReplicationABSTRACT
The cyclotides are a family of cyclic "mini" proteins that occur in Violaceae, Rubiaceae and Cucurbitaceae plant families and contain a head-to-tail cyclic backbone and a cystine knot arranged by three disulfide bonds. To study the natural cyclotides of V tianshanica, dried herb was extracted with 50% ethanol, and the concentrated aqueous extract was subjected to a solvent-solvent partitioning between water and hexane, ethyl acetate and n-butanol, separately. The n-butanol extract containing cyclotides was subjected to column chromatography over Sephadex LH-20, eluted with 30% methanol. The subfractions were directly reduced by DTT and analyzed by reverse-phase HPLC. The peaks with different retention times were shown on the profile of RP-HPLC and collected. The cyclotides were speculated based on masses range from 3 000 to 3 500 Da. The purified cyclotides were reduced with DTT, alkylated with iodoacetamide, and then were cleaved with endoproteinase Glu-C, endoproteinase Lys-C and Trypsin, separately. The digested peptides were purified on RP-HPLC and analyzed on MALDI TOF/TOF analyzer. A new cyclotide, cycloviolacin T1 and a reported cyclotide varv E were systemically determined using MALDI TOF/TOF system. So the method for the isolation and characterization of cyclotides was quickly built up in succession.
Subject(s)
Amino Acid Sequence , Chromatography, High Pressure Liquid , Cyclotides , Chemistry , Molecular Sequence Data , Molecular Structure , Plants, Medicinal , Chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tandem Mass Spectrometry , Viola , ChemistryABSTRACT
Overexpression of DNA methyltransferases(DNMTs)is observed in many tumors. Overexpressed DNMTs silence tumor suppressor genes by catalyzing the methylation of CpG islands within the promoter regions, and therefore promote the neoplastic transformation of normal cells. Highly elevated DNMT activities have been observed in cancers including gastrointestinal cancers, breast cancer, and lung cancer and have been associated with cancer development and prognosis. Drugs and targeted therapies that inhibit DNMT activities can reactivate methylated tumor suppressor genes, promote apoptosis, and thus inhibit tumor growth.
ABSTRACT
Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.