Simultaneous splicing of multiple DNA fragments in one PCR reaction

BACKGROUND: Rapid and simultaneous splicing of multiple DNA fragments is frequently required in many recombinant DNA projects. However, former overlap extension PCRs, the most common methods for splicing DNA fragments, are not really simultaneous fusing of multiple DNA fragments. RESULTS: We performed an optimized method which allowed simultaneous splicing of multiple DNA fragments in one PCR reaction. Shorter outermost primers were prior mixed with other PCR components at the same time. A sequential thermo cycling program was adopted for overlap extension reaction and amplification of spliced DNA. Annealing temperature was relatively higher in the overlap extension reaction stage than in the fused DNA amplification. Finally we successfully harvested target PCR products deriving from fusion of two to seven DNA fragments after 5–10 cycles for overlap extension reaction and then 30 cycles for fused DNA amplification. CONCLUSIONS: Our method provides more rapid, economical and handy approach to accurately splice multiple DNA fragments. We believe that our simultaneous splicing overlap extension PCR can be used to fuse more than seven DNA fragments as long as the DNA polymerase can match

The Effectiveness of Tepid Sponge Bath with 32oc and 37oc to Decrease Body Temperature at Toddler with Fever

Introduction: Tepid sponge bath is a therapeutic bath by washing all around of the body with warm water to decrease body temperature. Warm water that used were 32oC (nail warm) and 37oC (warm). The aimed of this study was to compare the effectivity of tepid sponge bath with 32oC and 37oC warm water on decreasing body temperature at toddler with fever. Method: A quasy experimental pre post test design was used in this study. The population was toddler who had body temperature ≥38oC which treated in anggrek pediatric room dr. Iskak public hospital Tulungagung. There were 26 respondents recruited by using purposive sampling technique and divided into two group, each of 13 respondents received tepid sponge bath with 32oC and others received tepid sponge bath with 37oC warm water. The independent variable was tepid sponge bath and dependent variable was body temperature. Data were collected by using digital termometere and noted in respondent observation, then analyzed by using Paired t-Test and Mann Withney U-Test. Result: The result showed that there was an effectivity on decreasing body temperature by giving tepid sponge bath with 32oC and 37oC warm water with significance level p=0.000 and there was a difference decreasing body temperature among both of them with significance level p=0.016. Discussion: It can be concluded that tepid sponge bath with 37oC warm water was more effective than tepid sponge bath with 32oC warm water. Further studies should be observed the effectivity of tepid sponge bath with more specific age, fever character and more time and respondent

Minimal clinically important difference of the EORTC QLQ-CIPN20 for worsening peripheral neuropathy in patients receiving neurotoxic chemotherapy.

Context/objectivesThis is the first study to determine the minimal clinically important difference (MCID) of the European Organisation of Research and Treatment of Cancer Quality of Life Questionnaire-CIPN twenty-item scale (EORTC QLQ-CIPN20), a validated instrument designed to elicit cancer patients' experience of symptoms and functional limitations related to chemotherapy-induced peripheral neuropathy.MethodsCancer patients receiving neurotoxic chemotherapy completed EORTC QLQ-CIPN20 and the Functional Assessment of Cancer Therapy/Gynecologic Oncology Group-Neurotoxicity [FACT/GOG-NTX] at baseline, second cycle of chemotherapy (T2, n = 287), and 12 months after chemotherapy (T3, n = 191). Anchor-based approach used the validated FACT/GOG-NTX neurotoxicity (Ntx) subscale to identify optimal MCID cutoff for deterioration. Distribution-based approach used one-third standard deviation (SD), half SD, and one standard error of measurement of the total EORTC QLQ-CIPN20 score.ResultsThere was a moderate correlation between the change scores of the Ntx subscale and sensory and motor subscales of QLQ-CIPN20 (T2: r = - 0.722, p < 0.001 and r = - 0.518, p < 0.001, respectively; T3: r = - 0.699; p < 0.001 and r = - 0.523, p < 0.001, respectively). The correlation between the change scores of the Ntx subscale and the QLQ-CIPN20 autonomic subscale was poor (T2: r = - 0.354, p < 0.001; T3: r = 0.286, p < 0.001). Based on the MCID derived using distribution-based method, the MCID for the QLQ-CIPN20 sensory subscale was 2.5-5.9 (6.9% to 16.4% of the subdomain score) and for motor subscale was 2.6-5.0 (8.1%-15.6% of the subdomain score).ConclusionThe MCID for the EORTC QLQ-CIPN20 established using distribution-based approaches was 2.5-5.9 for the sensory subscale and 2.6-5.0 for the motor subscale. When noted in assessments even with small change in scores, clinicians can be alerted for appropriate intervention

Anatomical features, fiber morphological, physical and mechanical properties of three years old new hybrid Paulownia: green Paulownia

Objective: Green Paulownia (hybridization of Paulownia elongata × Paulownia fotunei and tropical Paulownia spp.) is new hybrid claimed as one of the fast-growing woody plants with the high potential as a fiber material or lignocellulosic material. The material for this study originates from the area of Nanning in China. Methodology: Cell morphology and anatomical appearances were observed and evaluated under the image analysis system (Leica DMLS). Physical and mechanical properties were evaluated based on the American Society for Testing and Materials (ASTM) standards. Results: From the results, average value of the mean fiber length was 0.905 mm, mean fiber length 34.59 μm, lumen thickness 26.80 μm and cell wall thickness 3.89 μm. Fiber dimensions of green Paulownia are in the normal range for hardwoods. The physical and mechanical properties of 3 years old green Paulownia have similar properties than those 7-11 years old Paulownia published in China. Conclusion: The 3 years old green Paulownia timbers can be used as materials for furniture

A STUDY ON THE MOTIVATION OF HIGH SCHOOL VOCATIONAL SPORTS STUDENTS TO ATTEND YUAN ZE UNIVERSITY AND THEIR PLANNING FOR FURTHER STUDIES

The purpose of this study was to investigate the motivation of students in high school vocational physical education classes in Taoyuan City towards the athletic excellence of Yuan Ze University and their plans for further education and to analyze the influence of these factors on students' choices for further education. The target population of the study was students with physical education classes in public and private high schools in Taoyuan City, and the data were collected through a questionnaire survey. 200 questionnaires were distributed, and 150 valid questionnaires were eventually collected, with a recovery rate of 93%. As a result, 68.6% of the participants were male and 31.4% were female. ii. In terms of academic performance, the ranking of students was mainly concentrated in the top five. In terms of academic motivation, the majority of the students had a positive attitude towards their studies, with 67.8% of the students being average. In terms of the source of information for the athletic performance test, teachers and coaches recommended the most students, totaling 110 students, accounting for 72.4%. The main reason why students chose to study at Yuan Ze University was the recommendation from teachers and coaches. After entering Yuan Ze University, most students 58 students or 38.2%, planned to continue practicing and continue their athletic performance. The most popular course was the Bachelor of Management, with 45 students, 29.8%. It is an important reference value for the future counseling of athletes on further education and the formulation of related policies. It is expected to provide athletes with more comprehensive support and guidance on further education and to help athletes find a balance between sports and academics, and make wise choices in further education.  Article visualizations

Outreach syphilis testing services by different health providers to female sex workers in southern China.

Health providers have played important roles on delivering prevention and care services to control syphilis in China. The current study was aimed to evaluate the performance of different health providers in providing outreach syphilis testing services to female sex workers (FSWs). The current study carried out during April to August 2009 in Liuzhou was aimed to investigate the services delivered by two different types of clinics in China. A total of 1,808 FSWs recruited from sex work venues were included in the study. Prevalence of positive syphilis test (6.4%) among FSWs accessed by the local center for disease control outreach teams (CDC teams) was significantly lower than that (9.3%) among FSWs accessed by the local reproductive health hospital outreach teams (RHH teams). As compared with CDC teams, RHH teams had more FSWs to be successfully referred to the designated STD clinics for further syphilis confirmation and intervention (85.7% vs. 26.7%, P<0.001). These findings indicate that RHH teams may be more efficient than CDC teams to provide outreach-based services to FSWs. Participation of the reproductive health providers or other medical facilities in outreach services to FSWs should be considered in developing intervention programs in China

Replication and Meta-analysis of the Association between BDNF Val66Met Polymorphism and Cognitive Impairment in Patients Receiving Chemotherapy.

Cancer-related cognitive impairment (CRCI) adversely affects cancer patients. We had previously demonstrated that the BDNF Val66Met genetic polymorphism is associated with lower odds of subjective CRCI in the multitasking and verbal ability domains among breast cancer patients receiving chemotherapy. To further assess our previous findings, we evaluated the association of BDNF Val66Met polymorphism with subjective and objective CRCI in a temporally separate cohort of patients and pooled findings from both the original (n = 145) and current (n = 193) cohorts in a meta-analysis. Subjective CRCI was assessed using FACT-Cog. Objective CRCI was evaluated using computerized neuropsychological tests. Genotyping was carried out using Sanger sequencing. The association of BDNF Val66Met genotypes and CRCI was examined with logistic regression. A fixed-effect meta-analysis was conducted using the inverse variance method. In the meta-analysis (n = 338), significantly lower odds of CRCI were associated with Met allele carriers based on the global FACT-Cog score (OR = 0.52, 95% CI 0.29-0.94). Furthermore, Met allele carriers were at lower odds of developing impairment in the domains of memory (OR = 0.34, 95% CI: 0.17-0.70), multitasking (OR = 0.33, 95% CI: 0.18-0.59), and verbal ability (OR = 0.46, 95% CI: 0.24-0.88). Consistent with the previous study, lower odds of subjective CRCI among patients with the BDNF Met allele was observed after adjusting for potential confounders in the multitasking (OR = 0.30, 95% CI: 0.14-0.67) domain. In conclusion, carriers of the BDNF Met allele were protected against global subjective CRCI, particularly in the domains of memory, multitasking, and verbal ability. Our findings further contribute to the understanding of CRCI pathophysiology

STI571 reduces TRAIL-induced apoptosis in colon cancer cells: c-Abl activation by the death receptor leads to stress kinase-dependent cell death

<p>Abstract</p> <p>Background</p> <p>In an effort to achieve better cancer therapies, we elucidated the combination cancer therapy of STI571 (an inhibitor of Bcr-Abl and clinically used for chronic myelogenous leukemia) and TNF-related apoptosis-inducing ligand (TRAIL, a developing antitumor agent) in leukemia, colon, and prostate cancer cells.</p> <p>Methods</p> <p>Colon cancer (HCT116, SW480), prostate cancer (PC3, LNCaP) and leukemia (K562) cells were treated with STI571 and TRAIL. Cell viability was determined by MTT assay and sub-G1 appearance. Protein expression and kinase phosphorylation were determined by Western blotting. c-Abl and p73 activities were inhibited by target-specific small interfering (si)RNA. In vitro kinase assay of c-Abl was conducted using CRK as a substrate.</p> <p>Results</p> <p>We found that STI571 exerts opposite effects on the antitumor activity of TRAIL. It enhanced cytotoxicity in TRAIL-treated K562 leukemia cells and reduced TRAIL-induced apoptosis in HCT116 and SW480 colon cancer cells, while having no effect on PC3 and LNCaP cells. In colon and prostate cancer cells, TRAIL caused c-Abl cleavage to the active form via a caspase pathway. Interestingly, JNK and p38 MAPK inhibitors effectively blocked TRAIL-induced toxicity in the colon, but not in prostate cancer cells. Next, we found that STI571 could attenuate TRAIL-induced c-Abl, JNK and p38 activation in HCT116 cells. In addition, siRNA targeting knockdown of c-Abl and p73 also reduced TRAIL-induced cytotoxicity, rendering HCT116 cells less responsive to stress kinase activation, and masking the cytoprotective effect of STI571.</p> <p>Conclusions</p> <p>All together we demonstrate a novel mediator role of p73 in activating the stress kinases p38 and JNK in the classical apoptotic pathway of TRAIL. TRAIL via caspase-dependent action can sequentially activate c-Abl, p73, and stress kinases, which contribute to apoptosis in colon cancer cells. Through the inhibition of c-Abl-mediated apoptotic p73 signaling, STI571 reduces the antitumor activity of TRAIL in colon cancer cells. Our results raise additional concerns when developing combination cancer therapy with TRAIL and STI571 in the future.</p

Letter of Intent: Jinping Neutrino Experiment

Jinping Neutrino Experiment (Jinping) is proposed to significantly improve measurements on solar neutrinos and geoneutrinos in China Jinping Laboratory - a lab with a number of unparalleled features, thickest overburden, lowest reactor neutrino background, etc., which identify it as the world-best low-energy neutrino laboratory. The proposed experiment will have target mass of 4 kilotons of liquid scintillator or water-based liquid scintillator, with a fiducial mass of 2 kilotons for neutrino-electron scattering events and 3 kilotons for inverse-beta interaction events. A number of initial sensitivities studies have been carried out, including on the transition phase for the solar neutrinos oscillation from the vacuum to the matter effect, the discovery of solar neutrinos from the carbon-nitrogen-oxygen (CNO) cycle, the resolution of the high and low metallicity hypotheses, and the unambiguous separation on U and Th cascade decays from the dominant crustal anti-electron neutrinos in China.Comment: Proposal for the Jinping Neutrino Experimen