Toxicity and toxicokinetic assessment of an anti-tubercular drug pretomanid in cynomolgus monkeys.

Pretomanid is a nitroimidazooxazine antimycobacterial drug that was approved in more than 10 countries as part of a three-drug, all oral regimen, consisting of bedaquiline, pretomanid, and linezolid (BPaL) for 6-months treatment of adults with pulmonary extensively drug-resistant tuberculosis (XDR-TB) or with complicated forms of multidrug-resistant tuberculosis (MDR-TB). The toxicological profile of pretomanid was thoroughly evaluated in repeat-dose oral toxicity studies up to 39 weeks long in cynomolgus monkeys. Exposures up to 10-fold higher than in humans at the approved pretomanid dose (200 mg) were achieved in acute studies allowing for characterization of dose-limiting toxicity. Target organs and processes identified in acute and chronic toxicity studies included QT prolongation, nervous system effects, and liver effects (minimal hepatocellular hypertrophy without elevations in liver enzymes). In a 13-week study, no cataracts were present at the end of dosing, but 2 of 12 monkeys had cataracts at the end of a 13-week recovery period. No cataracts related to pretomanid administration were observed in subsequent 13-week or 39-week studies. No male reproductive toxicity was observed in these studies. No-observed-adverse-effect levels (NOAELs) were identified in all studies. Exposures at the NOAELs equaled, or exceeded, human exposure at the approved pretomanid dose with the exception of female monkeys in a 39-week chronic toxicity study. These data support the use of pretomanid as part of the 6-month BPaL regimen for treating XDR-TB and MDR-TB.

[QUADAS-C-A tool for assessing risk of bias regarding Quality Assessment of Diagnostic Accuracy Studies-Comparative].

This paper introduced the Quality Assessment of Diagnostic Accuracy Studies-Comparative (QUADAS-C), illustrated the comparison with the QUADAS-2, and using QUADAS-C together with QUADAS-2 to present QUADAS-C results through systematic reviews. Like the domain for QUADAS-2, QUADAS-C retained four domains, including patient selection, index test, reference standard, flow, and timing, and comprised additional questions for each QUADAS-2 part. Unlike the QUADAS-2 tool, the starting question of each domain for QUADAS-C was designed to summarize the risk of biased information captured by QUADAS-2. QUADAS-C only dealt with the risk of bias but did not include the part of concerns regarding applicability. The answers to signaling questions for each domain of QUADAS-C would lead to a 'low''high' or 'unclear' risk of biased judgment for the original study.

[Quality assessment of global breast cancer screening guidelines].

Objective: To evaluate the quality of the published breast cancer screening guidelines to provide a reference for domestic studies in the future. Methods: PubMed, Embase, Cochrane Library, Web of Science, SinoMed, China National Knowledge Infrastructure, VIP, and Wanfang Data were searched to identify breast cancer screening guidelines on until August 2020. Two reviewers screened literature and extracted data independently. The Appraisal of Guidelines for Research & Evaluation Ⅱ(AGREEⅡ) and Reporting Items for Practice Guidelines in Healthcare(RIGHT) tools were used to evaluate the quality of the included guidelines. Results: A total of 15 breast cancer screening guidelines were included, of which seven were published in the United States, with publication years focusing on 2015 to 2019, and 11 guidelines had updated versions. "Rigour of development" (47.0%±22.1%) and "Applicability" (44.0%±15.1%) of AGREEⅡ scored lower than other domains. "Review and quality assurance" (46.7%±39.9%) and "Funding, declaration, and management of interests" (41.7%±24.4%) of RIGHT were reported poorer than others. There were six guidelines recommended and another nine recommended with modifications based on the overall AGREEⅡ score. There were four guidelines with a good level, and another 11 were with a moderate level of RIGHT. The National Comprehensive Cancer Network published the best overall quality guidelines in 2018 (AGREEⅡ: 83.3%, RIGHT: 80.0%) and by the American Cancer Society in 2015 (AGREEⅡ: 83.3%, RIGHT: 85.7%). Conclusion: The quality of breast cancer screening guidelines was predominantly of moderate quality, and greater attention should be paid to the guideline development process and quality control of the guidelines.

[Quality assessment of global colorectal cancer screening guidelines and consensus].

Objective: To systematically evaluate the methodology and reporting quality of colorectal cancer (CRC) screening guidelines/consensus and provide lights for drafting CRC screening guidelines in China. Methods: The literature retrieval for all the Chinese and English guidelines published before September 1st, 2020 was conducted by using Chinese/English databases, such as China National Knowledge Infrastructure, Wanfang Data, VIP, SinoMed, PubMed, Embase, Web of Science, Cochrane Library, Guideline International Network, and supplement with the official website of multiple regions, such as the U.S. Preventive Services Task Force and American Cancer Society. We utilized The Appraisal of Guidelines for Research & Evaluation Ⅱ (AGREE Ⅱ) and Reporting Items for Practice Guidelines in Healthcare (RIGHT) approaches to assess the quality of CRC screening guidelines/consensus comprehensively. Results: After quality control, a total of 19 guidelines/consensus released by the United States, China, Australia, Canada, Britain, South Korea, and International organizations are successfully included, and strikingly, most of those belong to the United State(7). The results of the AGREE Ⅱ quality evaluation show that the average scores of scope and purpose (87.5%) and clarity of presentation (89.6%) are high. In contrast, there are deficient in stakeholder involvement (47.0%), the rigor of development (42.3%), applicability (47.5%), and editorial independence (50.2%). Among all the guidelines, there are 12 with an overall score of 50 or more, 13 with a recommendation level of "A", 2 with a rating of "B" and 4 with a rating of "C". Additionally, the RIGHT evaluation revealed that the average report rate in each field is necessary information (76.3%), background (77.0%), evidence (55.8%), recommendations (59.4%), review and quality assurance (26.3%), funding and declaration and management of interests (43.4%), other information (49.1%). Among all the guidelines, six have good reporting quality, whereas the additional 13 have general or weak evidence. Furthermore, subgroup analysis indicates that the quality of guidelines in developed countries is superior to that of China. Conclusion: The number of CRC screening guidelines/consensus is increasing gradually, and the overall quality of those is high, but the normative nature is warranted to be strengthened.

[The current situation of occupational burnout and its influencing factors among orphan child care workers in Nanjing].

Objective: To explore the status of job burnout of nurses for orphans and disabled children in Nanjing and its influencing factors, so as to provide reference for promoting the mental health of nurses for orphans and disabled children. Methods: From February to May, 2017, a cross-sectional survey was conducted among 236 nurses working in social welfare institutions for orphans and disabled children in Nanjing, using Occupational Stress Indicator (OSI) and Maslach Burnout Inventory (MBI) . It investigated the status of job burnout among nurses of orphans and disabled children, logistic regression analysis was used to explore the effect of occupational stress on the incidence of job burnout of nurses for orphans and disabled children. Results: The results showed that the incidence of job burnout was 67.37% (159/236) , the incidence of job burnout was negatively correlated with behavioral characteristics and family support, the incidence of emotional exhaustion was positively correlated with task control and job monotony (P<0.05) , and negatively correlated with role ambiguity and job satisfaction (P<0.05) . The incidence of depersonalization was negatively correlated with job monotony and peer support (P<0.05) . Conclusion: The nurses of orphans and disabled children have serious job burnout. Occupational stress factors, personality characteristics and relieving factors have influence on the incidence of job burnout.

[Study on occupational stress response of orphans and disabled children care workers in Nanjing].

Objective: To explore the status quo of occupational stress and its influencing factors of nursing staff for orphaned and disabled children in Nanjing, and to put forward measures for adverse reactions to occupational stress, so as to ensure the psychological health of this occupational population. Methods: From February to May 2017, 236 nursing assistants for orphans and disabled children in Nanjing social welfare home were selected by cluster sampling method. The occupational stress index(occupational stress indictor, OSI) was used to investigate occupational stress response and occupational stress factors. Partial correlation and multiple linear regression were used for statistical analysis. Results: The scores of job satisfaction, mental health and depressive symptoms in occupational stress reaction were (43. 99±6. 83) , (36. 09±4. 59) and (17. 31±2. 44) re- spectively. In terms of job satisfaction, work monotony, logic and compound change were the contributing fac- tors (P<0. 05) , task strategy and task control were negative factors (P<0. 05) ; on mental health, opportunities were raised and participation in decision-making Self-esteem, technology utilization, environmental control, time management, task strategy and support of colleagues as contributing factors (P<0. 05) , ambition and role conflicts as negative factors (P<0. 05) ; on depression, work input, participation Decision-making, promotion opportunities and behavioral characteristics were protective factors (P<0. 05 ) . Conclusion: Occupational stress among caregivers of orphans and disabled children cannot be ignored. Occupational stress reaction is serious. Occupational stress factors should be reduced and individual stress coping ability should be enhanced.

[Clinical characteristics and economic burden of influenza among children under 5 years old, in Suzhou, 2011-2017].

Objective: To understand the clinical characteristics and economic burden of influenza-like illness (ILI) children aged 0-59 months in the outpatient settings in Suzhou, China, 2011-2017. Methods: From March 2011 to February 2017, we conducted a prospective surveillance program on ILI for children aged less than 5 years at Soochow University Affiliated Children's Hospital. Through standard questionnaires and follow-up survey via telephone, we collected information regarding the demographic characteristics, medical history, clinical symptoms and both direct and indirect costs associated with influenza, of the patients. We then compared clinical characteristics and economic burden of influenza A/H1N1, A/H3N2, and B infections among children with ILI. Results: We enrolled 6 310 patients with ILI from March 2011 to February 2017 and collected all their throat swabs. 791 (12.9%) of the swabs showed positive for influenza virus, including 88 (11.1%) subtype influenza A/H1N1, 288 (36.4%) subtype influenza A/H3N2, and 415(52.5%) type influenza B. The proportions of cough, rhinorrhea, wheezing, vomiting and convulsion in influenza-positive children were higher than those influenza-negative children. Except for the prevalence rates of cough (χ(2)=9.227, P=0.010), wheezing (χ(2)=7.273, P=0.026) and vomiting (χ(2)=8.163, P=0.017), other clinical symptoms appeared similar between the three viral subtypes. Among all the ILI children, the average total cost per episode of influenza was 688.4 Yuan (95%CI: 630.1-746.7) for influenza-negative children; 768.0 Yuan (95%CI: 686.8-849.3) for influenza-positive children and 738.3 Yuan (95%CI: 655.5-821.1) for influenza B. Children with influenza A/H1N1 spent much more than those with influenza A/H3N2 or influenza B in the total cost (χ(2)=7.237, P=0.028). Conclusion: Children infected influenza showed higher prevalence rates of cough, rhinorrhea, wheezing, vomiting and convulsion than those without influenza. Influenza A/H1N1 subtype caused heavier economic burden than the other two influenza subtypes.

[Assessment of psychological status of inpatients with head and neck cancer before surgery].

Objective: To investigate the prevalence and psychosocial characteristics in inpatients with head and neck cancer before surgery. Method: From September 2015 to December 2016, 237 consecutive inpatients with head and neck cancer who had been scheduled for surgery were prospectively enrolled in Department of Head and Neck Surgery of Shanxi Provincial Tumor Hospital. Mental health symptoms were systematically investigated using three psychological instruments: symptom checklist-90 (SCL-90), Zung self-rating anxiety scale (SAS) and Zung self-rating depression scale (SDS). SPSS 17.0 software was used to analyze the data. And he results of SCL-90, SAS and SDS were compared with the Chinese norm. For all statistical analyses, a P value <0.05 was considered statistically significant. Result: Of 237 patients, 228 (96.2%) completed all the questionnaire. The scores of SCL-90 (1.60±0.44), SAS (46.67±8.51)and SDS(47.50±11.43)in patients with head and neck cancer were significantly higher than those of Chinese norm (t=3.093, t=17.29, t=4.29 respectively and P=0.003, P=0.001, P=0.001 respectively). Positive proportion identified by three measure tools are 32.9%, 35.5% and 36.8% respectively. And 42 patients (18.4%) suffered from both anxiety and depression. The SCL-90 scores were significantly higher than those of the normal standard population, including dimension of somatization obsessive-compulsive, anxiety, hostility, phobic-anxiety and psychoticism (t=4.47, 3.04, 2.87, 2.58, 5.46, 4.15 respectively, all P<0.05). Conclusions: This study offers important information regarding psychological status in inpatients with head and neck cancer before surgery. Identifying these patients using proper screening instrument is of great important clinical implications for the early detection, management, and reduction of the distress associated with head and neck cancer.

An in vivo and in vitro assessment of the anti-inflammatory, antinociceptive, and immunomodulatory activities of Clematis terniflora DC. extract, participation of aurantiamide acetate.

AIM: Clematis terniflora DC. has been widely used as a traditional Chinese medicine for the treatment of tonsillitis, rheumatoid arthritis, and prostatitis. Despite its widespread use in China, there are currently no studies systematically examined its therapeutic effects and mechanism of action. As such, the present study was conducted to evaluate the anti-inflammatory, antinociceptive, and immunomodulatory effects of C. terniflora DC. using rodent and cellular models. METHODS: The anti-inflammatory properties of the 70% ethanol eluted fraction of the 70% ethanol extract of C. terniflora DC. (EECTD) were evaluated using the xylene-induced ear swelling test, the carrageenan-induced edema model, and the cotton pellet granuloma method. Its antinociceptive activities were determined using both the acetic acid-induced writhing test and the hot plate assay. In parallel, we conducted an in vitro assay in LPS-induced RAW264.7 cells to examine the anti-inflammatory effects of EECTD and its purified form, aurantiamide acetate (AA) on inhibition of nitric oxide (NO) and prostaglandin E2 (PGE2) release. RESULTS: EECTD (300mg/kg) significantly reduced the number of writhing, extended the pain response latency, and suppressed xylene-induced ear swelling. Each EECTD treatment group also had significant inhibition of cotton granulation formation in addition to reduced carrageenan-induced paw edema. EECTD was also shown to alleviate signs of inflammation in histopathological paw sections. However, it had a less noticeable effect on mouse ear swelling in the delayed type hypersensitivity test. A purified compound was isolated from EECTD and its structure was identified as AA. In vitro experimental results showed that both EECTD and AA were able to significantly inhibit the release of pro-inflammatory cytokines NO and PGE2 on LPS-induced RAW264.7 cells. CONCLUSION: These results suggest that EECTD has significant anti-inflammatory and antinociceptive activities, partially related to one of the active substances identified as AA. We hypothesize that these effects are related to its ability to inhibit the production of cytokines NO and PGE2. However, further work will be needed to determine its exact mechanism of action.

A novel lentivirus for quantitative assessment of gene knockdown in stem cell differentiation.

Loss of gene function is a valuable tool for screening genes in cellular processes including stem cell differentiation differentiation. However, the criteria for evaluating gene knockdown are usually based on end-point analysis and real-time, dynamic information is lacking. To overcome these limitations, we engineered a shRNA encoding LentiViral Dual Promoter vector (shLVDP) that enabled real-time monitoring of mesenchymal stem (MSC) differentiation and simultaneous gene knockdown. In this vector, the activity of the alpha-smooth muscle actin (αSMA) promoter was measured by the expression of a destabilized green fluorescent protein, and was used as an indicator of myogenic differentiation; constitutive expression of discosoma red fluorescent protein was used to measure transduction efficiency and to normalize αSMA promoter activity; and shRNA was encoded by a doxycycline (Dox)-regulatable H1 promoter. Importantly, the normalized promoter activity was independent of lentivirus titer allowing quantitative assessment of gene knockdown. Using this vector, we evaluated 11 genes in the TGF-ß1 or Rho signaling pathway on SMC maturation and on MSC differentiation along the myogenic lineage. As expected, knockdown of genes such as Smad2/3 or RhoA inhibited myogenic differentiation, while knocking down the myogenic differentiation inhibitor, Klf4, increased αSMA promoter activity significantly. Notably, some genes for example, Smad7 or KLF4 showed differential regulation of myogenic differentiation in MSC from different anatomic locations such as bone marrow and hair follicles. Finally, Dox-regulatable shRNA expression enabled temporal control of gene knockdown and provided dynamic information on the effect of different genes on myogenic phenotype. Our data suggests that shLVDP may be ideal for development of lentiviral microarrays to decipher gene regulatory networks of complex biological processes such as stem cell differentiation or reprogramming.

The conduct of in vitro studies to address time-dependent inhibition of drug-metabolizing enzymes: a perspective of the pharmaceutical research and manufacturers of America.

Time-dependent inhibition (TDI) of cytochrome P450 (P450) enzymes caused by new molecular entities (NMEs) is of concern because such compounds can be responsible for clinically relevant drug-drug interactions (DDI). Although the biochemistry underlying mechanism-based inactivation (MBI) of P450 enzymes has been generally understood for several years, significant advances have been made only in the past few years regarding how in vitro time-dependent inhibition data can be used to understand and predict clinical DDI. In this article, a team of scientists from 16 pharmaceutical research organizations that are member companies of the Pharmaceutical Research and Manufacturers of America offer a discussion of the phenomenon of TDI with emphasis on the laboratory methods used in its measurement. Results of an anonymous survey regarding pharmaceutical industry practices and strategies around TDI are reported. Specific topics that still possess a high degree of uncertainty are raised, such as parameter estimates needed to make predictions of DDI magnitude from in vitro inactivation parameters. A description of follow-up mechanistic experiments that can be done to characterize TDI are described. A consensus recommendation regarding common practices to address TDI is included, the salient points of which include the use of a tiered approach wherein abbreviated assays are first used to determine whether NMEs demonstrate TDI or not, followed by more thorough inactivation studies for those that do to define the parameters needed for prediction of DDI.

Assessment of autopsic samples of carotid atherosclerosis in the aged by intravascular ultrasound.

To assess the value of intravascular ultrasound in detecting carotid atherosclerosis, we compared the ultrasound images of 48 carotid artery segments from autopsies with their histological findings. The results showed that by intravascular ultrasonography one could distinguish between elastic and muscular tissues of arteries, determine the lesions of fibroelastosis and calcified plaques on arterial wall, and precisely measure the wall thickness, inner and outer diameter, luminal area and cross-sectional area of arteries with a high correlation between the data measured from ultrasonography and histological study (r values were 0.98, 0.97, 0.97, 0.96 and 0.96, respectively). This study suggests that intravascular ultrasound might be effectively used for morphological study and detection of atherosclerotic lesions in vivo.