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Nucleic acids in plant tissue lysates can be captured quickly by a cellulose filter paper and prepared for amplification after a quick purification. In this study, a published filter paper strip method was modified by sticking the filter paper on a polyvinyl chloride resin (PVC) sheet. This modified method is named EZ-D, for EASY DNA extraction. Compared with the original cetyl trimethylammonium bromide (CTAB) method, DNA extracted by EZ-D is more efficient in polymerase chain reaction (PCR) amplification due to the more stable performance of the EZ-D stick. The EZ-D method is also faster, easier, and cheaper. PCR analyses showed that DNA extracted from several types of plant tissues by EZ-D was appropriate for specific identification of biological samples. A regular PCR reaction can detect the EZ-D-extracted DNA template at concentration as low as 0.1 ng/μL. Evaluation of the EZ-D showed that DNA extracts could be successfully amplified by PCR reaction for DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved without the need for specialized equipment. As an optimized DNA purification method, EZ-D shows great advantages in application and can be used widely in laboratories where equipment is limited and rapid results are required.
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Objective:To investigate the long-term effects of metabolically healthy obesity on the risks of type 2 diabetes, cardiovascular disease events, and its mortality over a 23-year follow-up.Methods:Based on the results of an oral glucose tolerance test, there were 519 participants with normal glucose tolerance and 630 with newly diagnosed type 2 diabetes enrolled in 1986 and then given to assess the long-term clinical outcomes during the 23-year follow-up in Daqing. Metabolically healthy obesity was defined as the overweight and obese individuals with no metabolic abnormalities (diabetes, hypertension, hyperlipidemia). Finally, we identified 682 participants (350 with normal glucose tolerance and 332 with newly diagnosed diabetes). They were divided into five groups: 211 individuals with metabolically healthy normal weight (MHNW group), 58 with metabolically healthy overweight and obesity (MHO group), 81, 109, 223 were metabolically unhealthy overweight and obesity with hypertension (MUHO group), type 2 diabetes (MUDO group), hypertension and diabetes (MUHDO group). Incidences of type 2 diabetes, morbidity and mortality of cardiovascular disease were compared among these groups.Results:Over 23 years, instead of the morbidity and mortality of cardiovascular disease, the incidence of type 2 diabetes in MHO group was two times higher than in MHNW group ( 24.1%, 12.5/1 000 person years vs 10.9%, 5.2/1 000 person years, P=0.01), with an age, sex, and smoking history-adjusted hazard ratio ( HR) of 2.42 (95% CI 1.24-4.74, P=0.01). The morbidity and mortality of cardiovascular disease in the groups of overweight and obesity with metabolically unhealthy were higher than in MHNW group, and increased across the subjects with MUHO, MUDO, MUHDO ( P<0.05). Conclusion:Compared with metabolically healthy normal weight participants, the metabolically healthy obese group was at increased risk of type 2 diabetes but not cardiovascular disease events and its mortality. On the contrary, the overweight and obese groups with metabolic abnormalities had significant higher incidence of type 2 diabetes, morbidity and mortality of cardiovascular diseases.
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OBJECTIVE: To analyze the current status of core emergency response capability of nurses for systematically managing coronavirus disease(COVID-19) in intensive care units and to explore the related influencing factors. METHODS: A total of 100 nurses who systematically manage COVID-19 in the intensive care units from a class 3 grade A hospital in Wuhan City were selected as study subjects by convenience sampling method. The self-designed COVID-19 Core Emergency Response Capability Questionnaire for Nurses was used to investigate the status of COVID-19 core emergency response capability. RESULTS: The total score of the core emergency capability of the nurses averaged 133.1±18.1, and the scoring rate was 80.7%, which was at a high level. The scoring rates of each dimension from high to low were prevention ability, rescue ability and preparation ability, with the score rate of 85.2%, 81.7% and 71.0% respectively. The results of generalized linear regression analysis showed that the level of core emergency response capability of nurses was higher in high-risk departments such as respiratory department, infectious department, intensive care unit, outpatient department and emergency department than that of low-risk departments [partial regression coefficient(β) =11.18, P<0.01]. The levels of core emergency response capability of nurses who were married, once worked in the fever clinic, and were members of the National Health Emergency Rescue Team was higher(β were 8.28, 24.34 and 10.89, respectively, P<0.05). CONCLUSION: The core emergency response capability of nurses who systematically manage COVID-19 in the critical care units in Wuhan City is at a high level. Their core emergency response capability is affected by factors such as personal work experience, marital status, emergency work experience and trainings.
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Objective To explore the influence of lifestyle intervention on long-term diabetes in subjects with impaired glucose tolerance (IGT) returned to normal glucose tolerance (NGT) within 6 years.Methods A total of 577 subjects (aged 25-74 years old) with IGT in Daqing were enrolled and randomly assigned to control,and diet,exercise and diet plus exercise groups in a six-year intervention trial in 1986.Subjects who were non-diabetic at the end of the intervention were followed up for additional 14 years.Results Among all the subjects,41.38％ of them who had returned to NGT from IGT within 6 years maintained NGT status after 20 years,and had a lower incidence of diabetes than subjects maintained IGT status (46.55％ vs.75.25％).Of note,in the intervention group,the percentage of participants developed diabetes in the NGT subjects was significantly lower than that in the IGT group (43.71％ vs.76.25％) after 20 years.There was high long-term risk for diabetes in the IGT subjects after the adjustment of age,sex and baseline glucose (HR=1.81,95％CI 1.27-2.58,P=0.001),whereas in the non-intervention group,no significant difference could be viewed in long-term diabetic risk between subjects maintained IGT status and those returned to NGT (71.43％ vs.65.22％) after adjusting of the same confounders (HR=1.03,95％CI 0.45-2.35,P=0.94).Conclusions IGT subjects who had returned to NGT in early years had lower risk for future diabetes than those who remained IGT.However,this beneficial effect could only be viewed in the intervention group,but not in the non-intervention group.
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Objective To explore the effect of execution sheet for health education in chronic heart failure (CHF) patients. Methods About 50 CHF patients hospitalized from October 2015 to December 2015 were assigned as the control group and another 50 CHF patients hospitalized from January to March 2016 were set up as the experiment groups. The control group was instructed by the regular health education and the experiment group was instructed by the execution sheet for health education. Results The executive ability in the experiment group were stronger than that of control group (P<0.01). The level of self management among the patients in the experiment group was higher than those of the that control group. Conclusions The health education execution sheet increase the patients' self-management and strengthen nursing ability for discharged patients. It is worthy of popularization in clinical practice.
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Objectives: To evaluate the effect and safety of 14-day continuous subcutaneous insulin infusion (CSII) in type 2 diabetes patients with heart diseases. Methods: A total of 22 consecutive type 2 diabetes patients (history ≤ 5 years) with heart diseases treated in our hospital from 2011-03 to 2013-08 were studied. There were 20 male, and the with the mean age of patieuts (48.15 ± 9.80) years, all patients without standard hypoglycemic treatment before admission. The patients received 14-day CSII for enhanced treatment and the blood glucose level, insulin function and insulin sensitivity were compared before and after the treatment. Results: After CSII treatment, the blood glucose level was obviously decreased, fasting blood glucose (FBG) and postprandial blood glucose at 30, 60 and 120 min were improved, all P Conclusion: CSII may quickly relieve glucotoxicity and improve insulin sensitivity in type 2 diabetes patients with heart diseases. Some patients may alleviate drug burden in clinical practice.
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<p><b>OBJECTIVE</b>To explore the variety of the genetic polymorphism of eight Prunella germplasm resources by AFLP analysis.</p><p><b>METHOD</b>The amplified fragment length polymorphism (AFLP) tags were applied to screen out 32 selective amplification primer pairs, the amplified bands as original matrix were analyzed with NTSYS-PC software for the similarity between the Prunella germplasm and the construction of genetic phylogenetic tree.</p><p><b>RESULT</b>SDS extraction of genomic Prunella DNA showed a good quality, could meet the requirements of AFLP analysis. From 32 selective amplification primer pairs, 10 pairs with strong polymorphism, better band and higher resolution were used for the construction of the AFLP Prunella fingerprint, all eight Prunella germplasms were separated, they were divided into 3 categories.</p><p><b>CONCLUSION</b>Prunella germplasm resources are rich in genetic diversity, certain morphological characteristics and differences are associate with genotype.</p>
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Amplified Fragment Length Polymorphism Analysis , DNA, Plant , Genetic Variation , Polymorphism, Genetic , Prunella , Classification , GeneticsABSTRACT
Objective To investigate the effects of chronic high glucose on α-cells glucagon releasing in relation to insulin resistance induced by high glucose. Methods TC1-6 cells, an α-cell line, were incubated separately in DMEM containing high (25.0 mmol/L), medium (11.1 mmol/L) and low (5.5 mmol/L) concentrations of glucose for 1 to 5 days. The secretion and gene expression of glueagon were measured. When TC1-6 cells had been cultured for 5 days, three different concentrations of insulin were added for 6 h and then glucagon secretion was detected. Western blot was used for 1 and 3 days to confirm the effect of high glucose on phosphorylation of Akt in TC1-6 cells. Results (1) Exposure of TC1-6 cells to 11.1 and 25.0 mmol/L glucose resulted in a slight increase of glucagon secretion compared with those incubated with 5.5 mmol/L. However, after 5 days in media containing 25.0 mmol/L glucose, glucagan secretion was significantly increased as compared to cells treated with low glucose [(136.80±10.94 vs 78.62±4.72 ) ng/106 cells, P<0.05]; moreover, in TC1-6 cell cultured with high glucose glucagon mRNA expression was increased significantly. (2) 10-7 mol/L insulin reduced significantly glucagon secretion of TC1-6 ceils exposed to low glucose [(21.59±1.30 vs 55.12±3.86) ng/106 cells], but just scarcely inhibited glucagon secretion of cells incubated with high glucose [(106.58±8.53 vs 117.18±10.55) ng/106 cells]. When insulin concentration was increased to 10-5 mol/L, glucagon secretion of TC1-6 cells in high glucose was also reduced [(46.55±3.72 vs 118.61±10.68 )ng/106 cells]. (3) After treated with 10-5 mol/L insulin for 2h, the levels of Akt phosphorylation in both groups of TC1-6 cells were increased by 180% and 70%, while the level in high glucose group was significantly lower than that in low glucose group. In the presence of phosphoinositide 3 kinase inhibitor, the levels of Akt phosphorylation were both lowered, but the inhibition in low glucose group was more significant than in high glucose group. Conclusion High glucose induces hypersecretion of glucagon, which may be due to the a-cell insulin resistance.