Cdc13 exhibits dynamic DNA strand exchange in the presence of telomeric DNA.

Telomerase is the enzyme that lengthens telomeres and is tightly regulated by a variety of means to maintain genome integrity. Several DNA helicases function at telomeres, and we previously found that the Saccharomyces cerevisiae helicases Hrq1 and Pif1 directly regulate telomerase. To extend these findings, we are investigating the interplay between helicases, single-stranded DNA (ssDNA) binding proteins (ssBPs), and telomerase. The yeast ssBPs Cdc13 and RPA differentially affect Hrq1 and Pif1 helicase activity, and experiments to measure helicase disruption of Cdc13/ssDNA complexes instead revealed that Cdc13 can exchange between substrates. Although other ssBPs display dynamic binding, this was unexpected with Cdc13 due to the reported in vitro stability of the Cdc13/telomeric ssDNA complex. We found that the DNA exchange by Cdc13 occurs rapidly at physiological temperatures, requires telomeric repeat sequence DNA, and is affected by ssDNA length. Cdc13 truncations revealed that the low-affinity binding site (OB1), which is distal from the high-affinity binding site (OB3), is required for this intermolecular dynamic DNA exchange (DDE). We hypothesize that DDE by Cdc13 is the basis for how Cdc13 'moves' at telomeres to alternate between modes where it regulates telomerase activity and assists in telomere replication.

Cdc13 exhibits dynamic DNA strand exchange in the presence of telomeric DNA.

Telomerase is the enzyme that lengthens telomeres and is tightly regulated by a variety of means to maintain genome integrity. Several DNA helicases function at telomeres, and we previously found that the Saccharomyces cerevisiae helicases Hrq1 and Pif1 directly regulate telomerase. To extend these findings, we are investigating the interplay between helicases, single-stranded DNA (ssDNA) binding proteins (ssBPs), and telomerase. The yeast ssBPs Cdc13 and RPA differentially affect Hrq1 and Pif1 helicase activity, and experiments to measure helicase disruption of Cdc13/ssDNA complexes instead revealed that Cdc13 can exchange between substrates. Although other ssBPs display dynamic binding, this was unexpected with Cdc13 due to the reported in vitro stability of the Cdc13/telomeric ssDNA complex. We found that the DNA exchange by Cdc13 occurs rapidly at physiological temperatures, requires telomeric repeat sequence DNA, and is affected by ssDNA length. Cdc13 truncations revealed that the low-affinity binding site (OB1), which is distal from the high-affinity binding site (OB3), is required for this intermolecular dynamic DNA exchange (DDE). We hypothesize that DDE by Cdc13 is the basis for how Cdc13 'moves' at telomeres to alternate between modes where it regulates telomerase activity and assists in telomere replication.

Physicians' Knowledge, Attitude and Practice of Generic Substitution in China: A Cross-Sectional Online Survey.

The purpose of this study is to investigate physicians' knowledge, attitudes and practice of generic medicine substitutions in China. We conducted a cross-sectional online questionnaire survey on physicians from secondary or tertiary hospitals in China from 2020 December to 2021 April. Descriptive statistical and ordered logistic regression were used for analysis. A total of 1225 physicians were included in the final analysis, and only 330 (26.94%) of them scored 4 or above in the knowledge part, which means that the physicians have a good knowledge of generic substitutions. Of the total, 586 (47.83%) agreed or strongly agreed that generic drugs could be substituted for originator drugs and 585 (47.75%) always or often prescribed generic medicines. The percentage of physicians with a positive attitude toward or that practice prescribing generic medicine is below 50%, which needs to be improved in China. Physicians' knowledge, their attitude toward generic substitution, if familiar with the policy of generic substitution, and incentives for prescribing generic medicines are influencing factors for the practice of generic substitution. Our studies show that the practice of generic substitution by physicians could be improved by several measures in China. We suggested that the physicians should be taught more about the bulk-buy policy and the generic-originator equivalence evaluation policy. Moreover, government incentives to promote generic substitution should be established. Our study also suggested that physicians with less working experience and female physicians should learn more about generic substitution.

Clinical Consumption of Compound Opioid Analgesics in China: A Retrospective Analysis of National Data 2015-2018.

Compound opioid analgesics (COA) are widely used for cancer pain relief, but few studies investigated the use of that. We aimed to report the characteristics and trend of COA consumption in different regions and health facilities in China. The procurement data of two types of COA, compound codeine phosphate (CCP) and oxycodone and acetaminophen (OAA), in all medical institutions of 20 provinces from 2015 to 2018 were used. Data were presented as defined daily dose for statistical purpose (SDDD) and expenditures per million inhabitants per day. The annual consumption of COA and ratio of two combinations were compared among regions and institutions. We found, during 2015-2018, COA consumption increased at an average rate of 7.32% in SDDD and 19.19% in expenditures, while OAA accounted for most of the consumption. Highest COA consumption appeared in Northern China, with 121.72 SDDD and 1689.87 RMB (2015), whereas the lowest COA consumption was only 11.28 SDDD appearing in Southern China. The ratio of OAA and CCP (in SDDD) was highest in Southern China (53.14 in 2018), whereas lowest in West North (0.37 in 2018). In terms of institutions, tertiary had the highest COA consumption, with 16.74 SDDD and 292.73 RMB (2018). The SDDD of OAA was 27.44 times of that of CCP in tertiary, while it was only 0.11 in primary. Overall, COA consumption is on an upward trend and different among regions and health institutions in either amount or types of COA. These findings call for establishment of COA management regulations.

Perceptions, Expectations, and Experience of Physicians About Pharmacists and Pharmaceutical Care Services in Pakistan: Findings and Implications.

Background: Optimal collaboration between pharmacists and other healthcare professionals such as physicians is integral in implementing pharmaceutical care. However, there are concerns regarding the role of pharmacists, especially among low- and middle-income countries. This study explored the perceptions, expectations, and experience of physicians working in various hospital settings of Punjab, Pakistan, about pharmacists and their roles. Methods: A self-administered questionnaire consisting of four sections was administered from October to December 2020. Descriptive and inferential statistics such as Kruskal-Wallis and Mann-Whitney tests were used for data analysis using SPSS. Results: Six hundred and seventy-eight physicians participated in this study with a response rate of 77.9%. Most of the physicians reported minimal to no interaction with pharmacists (n = 521, 76.8%). However, more than three-quarters of physicians (n = 660, 97.3%) accepted pharmacists as evidence-based sources of drug information. In addition, many physicians (n = 574, 84.7%) strongly agreed that pharmacists should attend patient care rounds to respond promptly to questions related to patient medication. A limited number of physicians (n = 124, 18.3%) assumed that pharmacists were advising their patients regarding the judicial use of their drugs. Median expectation and experience score had a significant association with age, experience, and education of physicians (P < 0.05). Conclusions: The perception of physicians was positive toward certain roles of pharmacists, coupled with high expectations. However, their experience was low, with most of the activities of pharmacists due to inadequate interprofessional coordination.

Impact of the Low-Price Medicine Policy on Medicine Supply in China: An Interrupted Time-Series Analysis.

Objectives: The primary objective of the study was to assess the impact of the Low-Price Medicine Policy (LPMP) on the supply of low-price medicines (LPMs) in China. The secondary objective of the study was to describe the supply situation of LPMs from 2005 to 2018. Methods: The LPMP was launched in the third quarter of 2014 (2014Q3). An interrupted time series analysis was used to evaluate the impact of LPMP on the supply of LPMs in China. Ordinary least squares and Poisson regression models were utilized to estimate the effect of LPMP on LPMs' supply growth rate and the number of supplied LPMs. All the LPMs were divided into two subgroups: intermittent supply and continuous supply. The trend and level changes of the quarterly average growth rate and number of quarterly supplies for different LPM groups were analyzed from 2005 to 2018. Findings: For the quarterly average growth rate, before the intervention, a significant increasing trend was observed in the total group and the continuous supply subgroup; after the introduction of LPMP, the increasing trend was ceased and a significant decrease in the trend and level was noted for both the total group (trend coefficient: ß 3 = -0.0132, p < 0.01; level coefficient: ß 2 = -0.1510, p < 0.05) and the continuous supply subgroup (trend coefficient: ß 3 = -0.0133, p < 0.01; level coefficient: ß 2 = -0.1520, p < 0.05); whereas it had no significant effect for intermittent supply subgroup. For the number of quarterly supplies, after the intervention of LPMP, decline of the supply number was observed (trend coefficient: ß 3 = -0.0027, p < 0.001; level coefficient: ß 2 = -0.0584, p < 0.001); whereas the LPMP was associated with an upward trend and level (trend coefficient: ß 3 = 0.0715, p < 0.001; level coefficient: ß 2 = 0.174) for the intermittent supply subgroup. Conclusion: For most of the LPMs, LPMP did not meet the goal of stimulating LPM production. However, for severely shortage medicines (the intermittent supply subgroup), the effect of LPMP was positive. Comprehensive policies rather than just deregulating medicine price should be introduced to alleviate the situation of medicine shortage in China.

Impact of video-led educational intervention on the uptake of influenza vaccine among adults aged 60 years and above in China: a study protocol for a randomized controlled trial.

BACKGROUND: Influenza is a global health threat to older adults, and the influenza vaccine is the most effective approach to prevent influenza infection. However, influenza vaccination coverage among Chinese older adults is far less than in developed countries such as the United States (4.0% vs. 64.9%). This study aims to increase influenza vaccination coverage in Chinese adults ≥60 years using a video-led educational intervention conducted by medical students. METHODS: A cluster randomized controlled trial will be conducted in 4 districts of Xi'an city, Shaanxi Province, China, using a stratified sampling approach. Adults aged ≥60 years will be recruited from 8 community hospitals. A self-administered questionnaire of knowledge, attitudes, and practices (KAP) will be employed to record the KAP score. During the 6-month interventional period, participants in the intervention group will receive educational videos focused on influenza and influenza vaccination, coupled with a group discussion conducted by the medical students. For those in the control group, no intervention will be provided. The outcomes measured in both groups will be the influenza vaccination coverage and the KAP scores of all participants. DISCUSSION: Medical students are more likely to educate older adults about scientific knowledge of influenza and its vaccine compared to clinical practitioners, who, most of the time, remain over-occupied due to the extensive workload. Video-led counseling and education could be a useful option to optimize older adults' understanding of influenza and influenza vaccination. This eventually could improve the uptake of influenza vaccine among Chinese older adults. TRIAL REGISTRATION: Chinese Clinical Trial Registry; ChiCTR2000034330 ; Registered 3rd July 2019.

Knowledge, attitude, and practices of community pharmacy staff toward antimicrobial stewardship programs: a cross-sectional study from Northeastern China.

BACKGROUND: Antimicrobial resistance is one of the most significant challenges of the twenty-first century, and the illegal sale of antimicrobial drugs at community pharmacies is a driver of antimicrobial resistance. This study explores the knowledge, attitudes, and practices (KAP) of pharmacy staff toward antimicrobial stewardship programs (ASPs). METHODS: We conducted a descriptive cross-sectional study among community pharmacy staff in Northeastern China, from April 1 to 31 May 2019, using a self-administered KAP questionnaire comprising 20 items. The data analysis was carried out by employing Mann-Whitney and Kruskal-Wallis tests. RESULTS: A response rate of 98.5% (394/400) was obtained. The majority of participants (94.9%) demonstrated a good understanding of antimicrobial use, but they lacked an adequate understanding of ASPs. Nearly half of the participants (40.6%) reported that they sold antimicrobials to patients without a prescription. Education level, age, occupation, and experience were all significantly associated (P < 0.05) with participants' median ASPs scores. Besides, the presence of a licensed pharmacist (OR 46.327, 95% CI 2.443-878.451, P = 0.011) was the main factor associated with the pharmacy staff's understanding of antimicrobial use policies. CONCLUSIONS: The participants' knowledge of antimicrobials was good, and their attitudes regarding ASPs were positive, but their practices regarding ASPs were poor.

Knowledge, Attitude, and Practices of Antibiotics and Antibiotic Resistance Among Chinese Pharmacy Customers: A Multicenter Survey Study.

BACKGROUND: Resistance to antibiotics is one of the major global health challenges. An adequate understanding of the public regarding rational antibiotic use is a prerequisite to limit progression in antibiotic resistance. This study aimed to investigate the knowledge, attitude, and practices (KAP) of antibiotics and antibiotic resistance among customers visiting community pharmacies. METHODS: This study was undertaken in three capital cities in China during March 2019 and July 2019 by using a questionnaire of 28 items. The questionnaire had four parts, including sociodemographic characteristics, KAP about antibiotics, and antibiotic resistance. A systematic random sampling approach was used to recruit the participants. Kruskal-Wallis and Mann-Whitney tests were carried out for data analysis. RESULTS: The response rate was 66.7% (1800/2700). Out of the total, only 9.7% of the customers had good knowledge about antibiotics. Nearly half of the participants were unable to differentiate between antibiotics, and anti-inflammatory drugs (n = 820, 45.6%, Median = 2, IQR = 1). Most of the customers were of the view that the use of over the counter antibiotics in pregnant women is unsafe (n = 1307, 72.6%, Median = 2, IQR = 0). Almost half of the participants disagreed that costly antibiotics are more effective and have fewer side effects (n = 897, 49.9%, Median = 3, IQR = 1). Only 22.3% of participants said that they always finish the course of antibiotic treatment (n = 401, 22.3%, Median = 3, IQR = 1). CONCLUSION: The knowledge of Chinese pharmacy consumers was inadequate, and a lack of good attitudes and practices in certain aspects of antibiotic use was observed. Educational interventions are needed to increase public knowledge of antibiotics.

Dispensing Antibiotics without a Prescription for Acute Cough Associated with Common Cold at Community Pharmacies in Shenyang, Northeastern China: A Cross-Sectional Study.

: The inappropriate use of antibiotics is a major health issue in China. We aimed to assess nonprescription antibiotic dispensing and assess pharmacy service practice at community pharmacies in Shenyang, northeastern China, and to compare these practices between pediatric and adult cases. A cross-sectional study was performed from March to May 2018 using the standardized client method. Two different simulated scenarios were presented at pharmacies, namely, pediatric and adult acute cough associated with a common cold. Of 150 pharmacy visits, 147 visits were completed (pediatric case: 73, adult case: 74). A total of 130 (88.4%) community pharmacies dispensed antibiotics without a prescription, with a significant difference between pediatric and adult cases (pediatric case, 79.5% versus adult case, 97.3%, p = 0.005). Symptoms were asked in most visits (pediatric case: 82.2%, adult case 82.4%). Patients' previous treatment and history of allergies were both inquired more frequently in the pediatric cases than in the adult cases. Medication advice was provided more often in the adult cases than in pediatric cases. Antibiotics were easily obtained without a prescription in Shenyang, especially for adult patients. Adequate inquiries and counseling had not occurred in most pharmacies.

O-Linked N-acetylglucosamine transferase 1 regulates global histone H4 acetylation via stabilization of the nonspecific lethal protein NSL3.

The human males absent on the first (MOF)-containing histone acetyltransferase nonspecific lethal (NSL) complex comprises nine subunits including the O-linked N-acetylglucosamine (O-GlcNAc) transferase, isoform 1 (OGT1). However, whether the O-GlcNAc transferase activity of OGT1 controls histone acetyltransferase activity of the NSL complex and whether OGT1 physically interacts with the other NSL complex subunits remain unclear. Here, we demonstrate that OGT1 regulates the activity of the NSL complex by mainly acetylating histone H4 Lys-16, Lys-5, and Lys-8 via O-GlcNAcylation and stabilization of the NSL complex subunit NSL3. Knocking down or overexpressing OGT1 in human cells remarkably affected the global acetylation of histone H4 residues Lys-16, Lys-5, and Lys-8. Because OGT1 is a subunit of the NSL complex, we also investigated the function of OGT1 in this complex. Co-transfection/co-immunoprecipitation experiments combined with in vitro O-GlcNAc transferase assays confirmed that OGT1 specifically binds to and O-GlcNAcylates NSL3. In addition, wheat germ agglutinin affinity purification verified the occurrence of O-GlcNAc modification on NSL3 in cells. Moreover, O-GlcNAcylation of NSL3 by wild-type OGT1 (OGT1-WT) stabilized NSL3. This stabilization was lost after co-transfection of NSL3 with an OGT1 mutant, OGT1C964A, that lacks O-GlcNAc transferase activity. Furthermore, stabilization of NSL3 by OGT1-WT significantly increased the global acetylation levels of H4 Lys-5, Lys-8, and Lys-16 in cells. These results suggest that OGT1 regulates the activity of the NSL complex by stabilizing NSL3.

Hepatotoxicity of benzotriazole and its effect on the cadmium induced toxicity in zebrafish Danio rerio.

As an emerging contaminant, 1-H-benzotriazole (1H-BTR) has been detected in the engineered and natural aquatic environments, which usually coexists with heavy metals and causes combined pollution. In the present study, wild-type and transgenic zebrafish Danio rerio were used to explore the acute toxicity as well as the single and joint hepatotoxicity of cadmium (Cd) and 1H-BTR. Although the acute toxicity of 1H-BTR to zebrafish was low, increased expression of liver-specific fatty acid binding protein was observed in transgenic zebrafish when the embryos were exposed to 5.0 µM of 1H-BTR for 30 days. Besides, co-exposure to 1H-BTR not only reduced the acute toxic effects induced by Cd, but also alleviated the Cd-induced liver atrophy in transgenic fish. Correspondingly, effects of combined exposure to 1H-BTR on the Cd-induced expressions of several signal pathway-related genes and superoxide dismutase and glutathione-s-transferase proteins were studied. Based on the determination of Cd bioaccumulation in fish and the complexing stability constant (ß) of Cd-BTR complex in solution, the detoxification mechanism of co-existing 1H-BTR on Cd to the zebrafish was discussed.

Lentiviral-mediated overexpression of KCTD12 inhibits the proliferation of human uveal melanoma OCM-1 cells.

Human potassium channel tetramerization domain containing 12 (KCTD12, also known as Pfetin) is a member of the KCTD family which consists of 26 members. It has been reported that KCTD12 regulates agonist potency and kinetics of GABAB receptor signaling. Proteomic analysis indicates that KCTD12 may be a potential biomarker for the diagnosis and prognosis of gastrointestinal stromal tumors. However, little has been reported concerning the role of KCTD12 in the other tumor types. In the present study, we designed and subcloned N-terminally Flag-tagged human KCTD12 into the pLVX­Puro vector. We then generated a human uveal melanoma cell line (OCM-1) stably expressing KCTD12. Using this stable cell line, we performed a series of experiments including colony formation, invasion, migration and wound-healing assays, flow cytometry and western blotting. Based on the experimental results, we first demonstrated that KCTD12 effectively suppressed the proliferation of OCM-1 cells and limited the spread of OCM-1 cells. In the flow cytometric analysis, prolongation of the progression of G2/M to G1 phase in the KCTD12-overexpressing OCM-1 cells was observed. In addition, inhibition of KCTD12-overexpressing OCM-1 cell xenograft growth in nude mice was observed. Taken together, KCTD12 may serve as a novel therapeutic target for patients with uveal melanoma.