Direct adenylation from 5'-OH-terminated oligonucleotides by a fusion enzyme containing Pfu RNA ligase and T4 polynucleotide kinase.

5'-Adenylated oligonucleotides (AppOligos) are widely used for single-stranded DNA/RNA ligation in next-generation sequencing (NGS) applications such as microRNA (miRNA) profiling. The ligation between an AppOligo adapter and target molecules (such as miRNA) no longer requires ATP, thereby minimizing potential self-ligations and simplifying library preparation procedures. AppOligos can be produced by chemical synthesis or enzymatic modification. However, adenylation via chemical synthesis is inefficient and expensive, while enzymatic modification requires pre-phosphorylated substrate and additional purification. Here we cloned and characterized the Pfu RNA ligase encoded by the PF0353 gene in the hyperthermophilic archaea Pyrococcus furiosus. We further engineered fusion enzymes containing both Pfu RNA ligase and T4 polynucleotide kinase. One fusion enzyme, 8H-AP, was thermostable and can directly catalyze 5'-OH-terminated DNA substrates to adenylated products. The newly discovered Pfu RNA ligase and the engineered fusion enzyme may be useful tools for applications using AppOligos.

Detection of DNA copy number alterations by matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of single nucleotide polymorphisms.

OBJECTIVES: Copy number alterations (CNAs) are frequently found in malignant tissues. Different approaches have been used for CNA detection. However, it is not easy to detect a large panel of CNA targets in heterogenous tumors. METHODS: We have developed a CNAs detection approach through quantitatively analyzed allelic imbalance by allelotyping single nucleotide polymorphisms (SNPs) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Furthermore, the copy number changes were quantified by real-competitive PCR (rcPCR) to distinguish loss of heterozygosity (LOH) and genomic amplification. The approach was used to validate the CNA regions detected by next generation sequencing (NGS) in early-stage lung carcinoma. RESULTS: CNAs were detected in heterogeneous DNA samples where tumor DNA is present at only 10% through the SNP based allelotyping. In addition, two different types of CNAs (loss of heterozygosity and chromosome amplification) were able to be distinguished quantitatively by rcPCR. Validation on a total of 41 SNPs from the selected CNA regions showed that copy number changes did occur, and the tissues from early-stage lung carcinoma were distinguished from normal. CONCLUSIONS: CNA detection by MALDI-TOF MS can be used for validating potentially interesting genomic regions identified from next generation sequencing, and for detecting CNAs in tumor tissues consisting of a mixture of neoplastic and normal cells.

Effect of family doctor contract services on patient perceived quality of primary care in southern China.

BACKGROUND: Family doctor contract service is an important service item in China's primary care reform. This research was designed to evaluate the impact of the provision of family doctor contract services on the patient-perceived quality of primary care, and therefore give evidence-based policy suggestions. METHODS: This cross-sectional study of family doctor contract service policy was conducted in three pilot cities in the Pearl River Delta, South China, using a multistage stratified sampling method. The validated Primary Care Assessment Tool-Adult Edition (PCAT-AS) was used to measure the quality of primary care services. PCAT-AS assesses each of the unique characteristics of primary care including first contact, continuity, comprehensiveness, coordination, family-centeredness, community orientation, culture orientation. Data was collected through face-to-face interviews held from July to November, 2015. Covariate analysis and multivariate Linear Regression were adopted to explore the effect of contract on the quality of primary care by controlling for the socio-demographic status and health care service utilization factors. RESULTS: A total of 828 valid questionnaires were collected. Among the interviewees, 453 patients signed the contract (54.7%) and 375 did not (45.3%). Multivariate linear regression showed that contracted patients reported higher scores in dimensions of PCAT total score (ß = - 8.98, P < 0.000), first contact-utilization(ß = - 0.71,P < 0.001), first contact-accessibility(ß = - 1.49, P < 0.001), continuity (ß = 1.27, P < 0.001), coordination (referral) (ß = - 1.42, P < 0.001), comprehensiveness (utilization) (ß = - 1.70, P < 0.001), comprehensiveness (provision) (ß = - 0.99, P < 0.001),family-centeredness(ß = - 0.52, P < 0.01), community orientation(ß = - 1.78, P < 0.001), than un- contracted after controlling socio-demographic and service utilization factors. There were no statistically significant differences in the dimensions of coordination (information system) (ß = - 0.25, P = 0.137) and culture orientation (ß = - 0.264, P = 0.056) between the two both groups. CONCLUSIONS: This study demonstrates that the pilot implementation of family doctor contract services has significantly improved patients' perceived primary care quality in the pilot cities, and could help solve the quality problem of primary care. It needs further promotion across the province.

Factors affecting turnover intentions among public hospital doctors in a middle-level city in central China.

Objective The aim of the present study was to explore prominent factors affecting turnover intentions among public hospital doctors in urban areas, particularly in Xiangyang City, Hubei Province, a middle-level city in central China. Methods Questionnaires were used to collect data from 284 public hospital doctors. Pearson's Chi-squared was used to assess whether sociodemographic and other factors were related to the turnover intentions of public hospital doctors. Binary logistic regression was performed to determine the significant factors that influence turnover intentions. Results The analysis revealed that 28.2% of public hospital doctors intended to leave the hospital where they were currently employed. Dissatisfaction with working conditions and hospital management processes, as well as work pressures, were significant factors contributing to the turnover intentions of public hospital doctors. Conclusion Research into turnover intentions indicates that public hospital doctors surveyed in urban China give greater weight to their professional environment and career development rather than salary in their employment decisions. What is known about the topic? Turnover of medical staff is a concern to hospital administrators because it is costly and detrimental to organisational performance and quality of care. Most studies have focused on the effects of individual and organisational factors on nurses' intentions to leave their employment. Income dissatisfaction was one of the determining factors of turnover intentions in previous studies. What does this paper add? The satisfaction of public hospital doctors with regard to income is not a determining factor of turnover intentions. In contrast with findings of previous studies, the doctors in public hospitals in urban China in the present study gave greater weight to their professional environment and career development in their employment decisions. What are the implications for practitioners? The findings suggest that health service managers and policy makers should pay greater attention to the factors affecting public hospital doctors' turnover intentions. This study will be useful for optimising public hospital management and minimising the turnover of doctors in China.

What is the job satisfaction and active participation of medical staff in public hospital reform: a study in Hubei province of China.

BACKGROUND: In China, public hospital reform has been underway for almost 5 years, and 311 pilot county hospitals are the current focus. This study aimed to assess the job satisfaction and active participation of medical staff in the reform. A total of 2268 medical staff members in pilot and non-pilot county hospitals in Hubei, China, were surveyed. METHODS: Questionnaires were used to collect data. The Pearson chi-square statistical method was used to assess the differences between pilot and non-pilot county hospitals and identify the factors related to job satisfaction as well as the understanding and perception of the reform. Binary logistic regression was performed to determine the significant factors that influence the job satisfaction of medical staff in pilot county hospitals. RESULTS: Medical staff members in pilot county hospitals expressed higher satisfaction on current working situation, performance appraisal system, concern showed by leaders, hospital management, and compensation packages (P < 0.05). They were exposed to work-related stress at a higher extent (P < 0.05) and half of them worked overtime. Within pilot county hospitals, less than half of the medical staff members were satisfied with current job and they have evidently less satisfaction on compensation packages and learning and training opportunities. The working hours and work stress were negatively related to the job satisfaction (P < 0.05). Satisfaction on the performance appraisal system, hospital management, compensation packages, and learning and training opportunities were positively related to job satisfaction (P < 0.05). Medical staff in pilot county hospitals exhibited better understanding of and more positive attitude towards the reform (P < 0.05). CONCLUSIONS: Pilot county hospitals have implemented some measures through the reform, but there still are deficiencies. The government officials and hospital administrators should pay attention to influencing factors of job satisfaction and focus on the reasonable demands of medical staff. In addition, the medical staff in pilot county hospitals exhibited a better understanding of the public hospital reform programme and showed more firm confidence, but there still were some medical staff members who hold negative attitude. The publicity and education of the public hospital reform still need improvement.

Factors influencing the work passion of Chinese community health service workers: an investigation in five provinces.

BACKGROUND: After the implementation of new healthcare reform, Chinese government paid increasing attention to developing community health service (CHS). The current focus is mainly on cultivating community general practitioners but paying less attention to the working status and occupational demands of in-service CHS workers. Work passion is playing an important role for medical workers. With work passion, CHS workers' team will become more stable and more effective, ensuring the sustainable development of CHS system. At present, the work passion of CHS workers is relatively low. Studying on influencing factors of work passion of CHS workers, promoting their work passion, and making them keep enthusiasm for work are significant. METHODS: A total of 100 CHS organizations were sampled randomly in 10 cities from 5 Chinese provinces for this study. A total of 3450 CHS workers from these CHS institutions took part in the surveys. Questionnaires were used to collect data, including socio-demographic information, work passion and opinion on influencing causes, and work-related satisfaction. Pearson chi-square statistical method was used to identify the factors related to CHS workers' work passion. Binary logistic regression was performed to determine the significant factors that influence CHS workers' work passion. RESULTS: A total of 38.77% of those who accomplished the questionnaire expressed that they didn't have passion for current work. The related factors that influence CHS workers' work passion are (1) socio-demographic factors such as age, and years of employment, and (2) other work-related factors such as learning and training opportunities, compensation packages, work stress, and personal development opportunities. CHS workers were most dissatisfied with the balance between remuneration and workload, job promotion opportunities. CONCLUSIONS: Based on the results, the government should concern for CHS workers' working status and work-related demands, pay more attention and meet their demands for reasonable compensation packages and self-development, balance the income and workload, provide more learning and training opportunities and personal development opportunities for CHS workers, in order to promote CHS workers' work satisfaction, improve their work passion and enthusiasm.

Factors influencing the turnover intention of Chinese community health service workers based on the investigation results of five provinces.

After the implementation of the new health reform, the Chinese government paid increasing attention to developing its community health service (CHS). The focus is mainly on cultivating community general medical practitioners but paying less attention to the working status and occupational demands of in-service CHS workers. CHS requires a stable team that can provide good service to community residents. At present, the demission rate of CHS workers is high. Studying how to increase the stability of CHS workers is noteworthy. The goal of this study is to ascertain the key factors that influence the CHS worker turnover intention to increase their work satisfaction and stability. A total of 100 CHS organizations were sampled randomly in 10 cities from 5 Chinese provinces for this study. All CHS workers from these organizations took a questionnaire survey. In total, 3,212 valid answer sheets were collected. Pearson Chi square test and Binary logistic regression were used to analyze the related influencing factors that result in CHS worker turnover intention. A total of 38.7% of those who accomplished the questionnaire intended to quit. The influencing factors that result in CHS worker turnover intention are (1) socio-demographic factors such as age, post of duty, professional title, and working seniority, and (2) other work-related factors such as pay packets, learning and training opportunities, promotion and personal development space, and working stress. CHS workers were less satisfied with the balance between payment and work quantity, promotion opportunity, and working conditions. Based on the results, the government should pay more attention to the various demands of CHS workers in service, especially by increasing their income, providing more learning and training opportunities, and increasing the degree of their work satisfaction to avoid turnover intention and ensure the stability of the CHS workforce.

Do patients choose community health services (CHS) for first treatment in China? Results from a community health survey in urban areas.

In recent years, the government has paid more attention to the development of community health service (CHS) in urban areas in China. Therefore, determining if it plays important roles and establishing methods to evaluate the effects of CHS are critical emphases in research. This study measured the effects of CHS through the choices of patients and their evaluation of CHS, and aimed to contribute to the development of primary health services. Face-to-face interviews were performed using the questionnaire with a random sample of 865 patients in CHS institutions from five provinces in China. Pearson's Chi square tests and binary logistic regression were used to analyze influencing factors that are associated with the patients' choices and evaluations. A total of 62.2 % of the patients would choose CHS for their first treatment. Patient choice was mainly affected by the following: (1) social demographic factors of the patients, namely, age, educational level, medical insurance, and survey areas; (2) evaluation of CHS by the patients: convenience, reasonable charges, and attitude of the doctors. In addition, the patients showed more satisfaction with convenience, waiting time, and communication with doctors, and less satisfaction with the medical charges, drug costs, and medical equipment of CHS. Through the results, we suggest that the government should provide more regard to the convenience, reasonable charges, and the attitude of the doctors as important factors to attract the patients to CHS. The government should also exert efforts to reduce the medical charges (especially the drug costs) for CHS.

The mediating role of negative emotions in the relationship between smoking and health-related quality of life among Chinese individuals: A cross-sectional study.

INTRODUCTION: Although the negative impact of smoking on health has been confirmed in various studies, few have explored psychological factors mediating the relationship between smoking and health-related quality of life (HRQOL). This study aimed to investigate the relationship between smoking and HRQOL in the Chinese population and the mediating role of negative emotions (NEs). METHODS: Survey data were derived from a cross-sectional study conducted in China from 20 June to 31 August 2022. We recruited participants from 148 cities across the country using a stratified multistage sampling method. The HRQOL of the dependent variable was measured using the Chinese version of European Quality of Life-5 Dimensions (EQ-5D-5L). The Patient Health Questionnaire (PHQ-9), Generalized Anxiety Disorder (GAD-7), and Perceived Stress Scale (PSS-4) were used to measure NE parameters including depression, anxiety, and perceived stress, as the intermediate variables. A multiple parallel mediation model was used to analyze the mediating role of NEs in smoking and HRQOL. RESULTS: A total of 21916 valid questionnaires were collected, of which 3010 (13.7%) and 18906 (86.3%) were categorized into smokers and non-smokers, respectively. The HRQOL (EQ-VAS score) of smokers (71.70 ± 23.08) was lower than that of non-smokers (73.69 ± 21.32), whereas the depression and anxiety levels of smokers were higher than those of non-smokers (all p<0.001). Moreover, smoking, NEs (depression and anxiety), and HRQOL showed pairwise correlations. According to the mediation analysis, depression (ß= -0.461; 95% BCa CI: -0.664 - -0.268) and anxiety (ß= -0.279; 95% BCa CI: -0.435 - -0.138) mediated the relationship between smoking and HRQOL after adjusting for demographic and life factors. CONCLUSIONS: These findings emphasize the necessity of studying the interaction between smoking, HRQOL, and Nes, and complementing the research on the impact of psychological factors on the HRQOL of smokers. Public health activities should focus on mental health and take targeted measures for the prevention, treatment, and rehabilitation of smokers.

Influence of social support on subjective well-being of patients with chronic diseases in China: chain-mediating effect of self-efficacy and perceived stress.

Introduction: The well-being of patients with chronic diseases is an issue of widespread concern in public health. While social support is thought to have a positive effect on it, the mechanisms of its influence have not been fully addressed. Thus, we explored the possible mediating effects of self-efficacy and perceived stress to determine the relationship between social support and well-being in these patients. Methods: A cross-sectional study was conducted among 4,657 patients with chronic diseases in China. The PROCESS Macro model 6 of SPSS was employed to explore the intermediary role between variables. Results: Self-efficacy and perceived stress played a partial intermediary role between social support and subjective well-being, with an effect ratio of 48.25% and 23.61%, respectively. Self-efficacy and perceived stress had a chain intermediary effect (28.14%) between social support and subjective well-being. Discussion: This study suggested that improving the self-efficacy of patients with chronic diseases to cope with the changes in social support caused by the disease could reduce stress and enhance subjective well-being.

Have the Pilot County Hospitals' Service Capability Been Improved Since the Healthcare Reform? An Analysis of 370 Hospitals in China.

BACKGROUND: County public hospital reform is one of the major tasks proposed in Chinese Healthcare Reform., and the evaluation of hospital reform effectiveness is very important and beneficial since it helps the government to understand the current situation of pilot county public hospitals and smoothly start the reform in all county hospitals. METHODS: This study used hospitals data from 2009 to 2012 to evaluate the effectiveness of county public hospital reform through comprehensive service capability. Descriptive analysis method was used, and factor analysis method was used to extract the main factors associated with service capabilities as well as to calculate a composite score. The t-test of two independent-samples methods was used to comparison analyze. RESULTS: The differences of common factor scores (hospital scale and service capacity, treatment quality, service quality, and services efficiency) between pilot and non-pilot hospitals were not statistically significant (P>0.05). The service capability score in 2012 was better than that in 2009 either in pilot or non-pilot group (P<0.05). The pilot hospitals' service capability score was better than that in non-pilot groups either in 2010 or 2012 (P<0.05). However, the differences from 2009 to 2012 of service capability score between pilot and non-pilot hospitals were not statistically significant. CONCLUSION: The comprehensive service capability of both pilot and non-pilot group all got improvement. However, county public hospital reform did not significantly play a due good role in improving the service capability in pilot group. The reform was helpful to improve the hospital current situation, but it has not completely achieved policy objectives in the sample hospitals of this study.

Is there an improvement of antibiotic use in China? Evidence from the usage analysis of combination antibiotic therapy for type I incisions in 244 hospitals.

The improvement of antibiotic rational use in China was studied by usage analysis of combination antibiotic therapy for type I incisions in 244 hospitals. Five kinds of hospitals, including general hospital, maternity hospital, children's hospital, stomatological hospital and cancer hospital, from 30 provinces were surveyed. A systematic random sampling strategy was employed to select outpatient prescriptions and inpatient cases in 2011 and 2012. A total of 29 280 outpatient prescriptions and 73 200 inpatient cases from 244 hospitals in each year were analyzed. Data were collected with regards to the implementation of the national antibiotic stewardship program (NASP), the overall usage and the prophylactic use of antibiotic for type I incisions. Univariate analysis was used for microbiological diagnosis rate before antimicrobial therapy, prophylactic use of antibiotics for type I incision operation, and so on. For multivariate analysis, the use of antibiotics was dichotomized according to the guidelines, and entered as binary values into logistic regression analysis. The results were compared with the corresponding criteria given by the guidelines of this campaign. The antibiotic stewardship in China was effective in that more than 80% of each kind of hospitals achieved the criteria of recommended antibiotics varieties. Hospital type appeared to be a factor statistically associated with stewardship outcome. The prophylactic use of antibiotics on type I incision operations decreased by 16.22% (P<0.05). The usage of combination antibiotic therapy for type I incisions was also decreased. Region and bed size were the main determinants on surgical prophylaxis for type I incision. This national analysis of hospitals on antibiotic use and stewardship allows relevant comparisons for bench marking. More efforts addressing the root cause of antibiotics abuse would continue to improve the rational use of antibiotics in China.