A novel V263I mutation in the glutamate-gated chloride channel of Plutella xylostella (L.) confers a high level of resistance to abamectin.

The frequent and extensive use of insecticides leads to the evolution of insecticide resistance, which has become one of the constraints on global agricultural production. Avermectins are microbial-derived insecticides that target a wide number of insect pests, including the diamondback moth Plutella xylostella, an important global pest of brassicaceous vegetables. However, field populations of P. xylostella have evolved serious resistance to avermectins, including abamectin, thereby threatening the efficiency of these insecticides. In this study, a novel valine to isoleucine mutation (V263I) was identified in the glutamate-gated chloride channel (GluCl) of field P. xylostella populations, which showed different levels of resistance to abamectin. Electrophysiological analysis revealed that the V263I mutation significantly reduced the sensitivity of PxGluCl to abamectin by 6.9-fold. Genome-modified Drosophila melanogaster carrying the V263I mutation exhibited 27.1-fold resistance to abamectin. Then, a knockin strain (V263I-KI) of P. xylostella expressing the homozygous V263I mutation was successfully constructed using the CRISPR/Cas9. The V263I-KI had high resistance to abamectin (106.3-fold), but significantly reduced fecundity. In this study, the function of V263I mutation in PxGluCl was verified for the first time. These findings provide a more comprehensive understanding of abamectin resistance mechanisms and lay the foundation for providing a new molecular detection method for abamectin resistance monitoring.

CRISPR/Cas9-mediated D472N substitution in the Rdl1 of Plutella xylostella confers low resistance to abamectin.

BACKGROUND: Abamectin is one of the main insecticides used for the control of Plutella xylostella, a destructive pest of cruciferous crops. Target-site mutation plays an important role in insecticide resistance. A point mutation (D472N) has been reported in the Rdl1 γ-aminobutyric acid receptor (GABAR) in P. xylostella, but its roles in insecticide resistance remain unknown. RESULTS: In this study, the D472N mutation of the Rdl1 GABAR was detected in several field populations of P. xylostella and showed a positive correlation with abamectin resistance. A knock-in homozygous mutation strain (D472N-KI) of P. xylostella was successfully constructed using CRISPR/Cas9 coupled with homology-directed repair, and the bioassay results demonstrated that compared with the susceptible strain, the D472N-KI strain had 11.1- and 3.7-fold increased resistance to abamectin and endosulfan, respectively. There was no difference in resistance to fipronil, broflanilide or isocycloseram, which also target the GABAR. In addition, the total fecundity of the D472N-KI strain was significantly reduced by 50.0%. CONCLUSION: Our results suggest that the homozygous D472N mutation in Rdl1 confers a low level of resistance to abamectin in P. xylostella but causes significant fecundity disadvantages, which may delay the development of resistance to some extent. These results lay a foundation for further understanding the mechanisms of abamectin resistance in insect pests. © 2022 Society of Chemical Industry.

A ceRNA network mediated by LINC00475 in papillary thyroid carcinoma.

Papillary thyroid carcinoma (PTC) is the most frequent histological type of differentiated thyroid carcinoma. Long noncoding RNAs (lncRNAs) have been widely reported to play a key role in human malignancies, and PTC is included. This study aimed to find out the functions and mechanism of lncRNA LINC00475 in PTC. LINC00475 was upregulated in PTC cells and was mainly located in the cytoplasm according to reverse-transcription polymerase chain reaction analyses and subcellular fractionation assays. As shown by cell counting kit-8 assays, ethynyl deoxyuridine incorporation assays, wound healing assays, and transwell assays, LINC00475 knockdown suppressed cell viability, proliferation, migration, and invasion. Mechanistically, LINC00475 upregulated the expression of messenger RNA zinc finger CCHC-type containing 12 (ZCCHC12) by binding to miR-376c-3p. ZCCHC12 was a direct target gene of miR-376c-3p in PTC cells. The relationship between miR-376c-3p and LINC00475 (or ZCCHC12) in PTC cells was probed by luciferase reporter assays, RNA pulldown assays, and RNA immunoprecipitation assays. In addition, both mRNA and protein levels of ZCCHC12 were downregulated due to miR-376c-3p overexpression or LINC00475 silencing. ZCCHC12 overexpression partially reversed the suppressive effect of LINC00475 knockdown on malignant behaviors of PTC cells. In conclusion, LINC00475 promotes PTC cell proliferation, migration, and invasion by upregulating ZCCHC12 via the interaction with miR-376c-3p.

PG545 alleviates diabetic retinopathy by promoting retinal Müller cell autophagy to inhibit the inflammatory response.

Diabetic retinopathy (DR), a major cause of blindness in working-age people, is attributed to the inflammatory response of retinal Müller cells (RMCs). The heparanase inhibitor PG545 plays proautophagic and anti-inflammatory roles. Intraperitoneal injection of PG545 at a dose of 20 mg/kg/d clearly reduced diabetes-induced body weight changes and fasting blood glucose levels in mice. PG545 also mitigated the reduction in retinal thickness and the formation of microaneurysms by promoting autophagy to inhibit the inflammatory response. In vitro, PG545 stimulated autophagy to downregulate the inflammatory response in high glucose-induced primary adult mouse RMCs. These data suggest that PG545 mitigates DR by promoting RMC autophagy to inhibit the inflammatory response.

The predictive value for excellent response to initial therapy in differentiated thyroid cancer: preablation-stimulated thyroglobulin better than the TNM stage.

PURPOSE: This study aimed to identify the predictive value of the low preablation-stimulated thyroglobulin (ps-Tg, <2 ng/ml) for excellent response to radioiodine remnant ablation in differentiated thyroid carcinoma. PATIENTS AND METHODS: A total of 398 consecutive patients who underwent total thyroidectomy and radioactive iodine remnant ablation therapy were reviewed retrospectively. Each patient was risk-stratified using the American Joint Cancer Committee and risk staging systems and using response to the initial therapy reclassification system. ps-Tg was defined as less than 2 ng/ml with negative thyroglobulin antibody under thyroid-stimulating hormone stimulation. A multivariate analysis was carried out for ps-Tg, TNM stage, and other potential clinical and pathologic factors. RESULTS: We followed the patients for a median of 32.7 months. Overall, an excellent rate of response was achieved in 367 (92.2%) of the 398 patients. The only variable found to be associated with excellent response was ps-Tg (odds ratio=2.530, P=0.009) by multivariate analysis. The subgroups with 0

Is papillary thyroid microcarcinoma an indolent tumor?: A retrospective study on 280 cases treated with radioiodine.

The increasing detection of papillary thyroid microcarcinoma (PTMC) has created management dilemmas. To clarify the clinical significance of postsurgery stimulated thyroglobulin (ps-Tg) in PTMC who undergo thyroidectomy and radioactive iodine (RAI), we retrospectively reviewed the 358 PTMC patients who were treated with RAI and followed up in our hospital. Those with an excessive anti-Tg antibody, ultrasound-detected residual were excluded, thereby resulting in the inclusion of 280 cases. Their clinical and histopathological information and clinical outcomes were collected and summarized. Tumor stages were classified according to the tumor, node, metastasis (TNM) staging system and the consensus of the European Thyroid Association (ETA) risk stratification system, respectively. Kaplan-Meier curves were constructed to compare the disease-free survival (DFS) rates of different risk-staging systems. By the end of follow-up, none of the patients died of the disease or relapsed. The 8-year DFS rate was 76.9%. Kaplan-Meier curves showed different DFS rates in TNM stages I versus IV, III versus IV, very low risk versus high risk, low risk versus high risk, respectively (P < 0.05), while they were not significantly different in stage I versus stage III, very low risk versus low risk (P > 0.05). Finally, 40 (14.3%) cases got a persistent disease. Five variables (male sex, nonconcurrent benign pathology, initial tumor size >5 mm, lymph node metastasis, and ps-Tg ≥ 10 µg/L) were associated with disease persistence by univariate regression analysis. Ps-Tg ≥ 10 µg/L was the only independent prognostic variable that predicted disease persistence by multivariate regression analysis (odds ratio: 36.057, P = 0.000). Therefore, PTMC with a small size of ≤1 cm does not always act as an indolent tumor. In conclusion, ps-Tg ≥ 10 µg/L is associated with increased odds of disease persistence. ETA risk stratification is more effective in predicting disease persistence than the TNM classification system.

Housing type and myopia: the mediating role of parental myopia.

BACKGROUND: Myopia has become a significant global public health concern, and is highly prevalent worldwide especially in Asian countries. It is associated with genetic factors as well as socioeconomic status; however, the underlying cause for school myopia has not been established. This study evaluates the impact of living environment on school myopia in Chinese school-aged children. METHODS: A large cross-sectional sample of area- and ethnicity-matched school children; a total of 43, 771 children from 12 cities participated in this study. The presence of myopia was self-reported and potential risk factors were determined by questionnaires. RESULTS: The self-reported prevalence of myopia in Chinese children was 31.8 % (n = 13, 928). In multiple logistic regression analysis, higer risk of myopia among school-aged children was significantly positively associated with both parental myopia (OR = 3.57; 95 % CI: 3.26-3.90), living in 1-3 floor (OR = 1.28; 95 % CI: 2.57-3.15), 4-6 floor (OR = 1.84; 95 % CI: 1.73-1.95) and 7 floor or more (OR = 2.02; 95 % CI: 1.88-2.16). Particularly, housing type was independently associated with myopia after stratified by parental myopia. An increasing prevalence of myopia was found with increasing floor of housing type in each outdoor time group. CONCLUSIONS: Housing type was independently associated with myopia, after stratified by parental myopia. Flat room, lower living floor and more outdoor time may be protective factors for myopia among school-aged children in mainland China.

[Analysis on the time of sunshine exposure among Chinese primary and middle school students in 12 provinces].

OBJECTIVE: To investigate the characteristics on the time of sunshine exposure in Chinese primary and middle school students. METHODS: A total of 43 771 primary and middle school students were randomly selected from 12 provinces in China. A self-reported questionnaire was used to understand their time of sunshine exposure on weekdays and weekends as well as related family information and behaviors.χ(2) test was used to compare the hours of sunshine exposure among different groups. Multivariate logistic regression analysis was conducted to identify the influencing factors. RESULTS: Results from the current survey indicated that the average time of sunshine exposure on weekdays and weekends were 2.49 and 3.22 hours, respectively. On weekdays, proportions of students with time of sunshine exposure ≤2 h/d or >2 h/d were 60.81% and 39.19%, respectively. On weekends, the proportions became 44.05% and 55.95%, respectively. On weekdays, the time of sunshine exposure as ≤2 h/d was significantly different among factors as sex, urban or rural area, residential area, self-rating academic performance, type of housing or ways of commuting to school. On weekends, the time of sunshine exposure ≤2 h/d were significantly different among sex, residential areas, self-rating academic performance, type of housing and ways of commuting to school. Data from the multivariate logistic regression analysis showed that the limited sunshine exposure was significantly associated with factors as: type of primary schools, being girls, students living in the urban area or living in central or western areas, with normal or better academic performances, living in a flat apartment and the time spent on watching video program >2 h/d both on weekdays and weekends. CONCLUSION: The time of sunshine exposure seemed not enough among school students and significant differences were seen among different groups.

Inhibition of BRD4 suppresses tumor growth and enhances iodine uptake in thyroid cancer.

Thyroid cancer is a common malignancy of the endocrine system. Although radioiodine (131)I treatment on differentiated thyroid cancer is widely used, many patients still fail to benefit from (131)I therapy. Therefore, exploration of novel targeted therapies to suppress tumor growth and improve radioiodine uptake remains necessary. Bromodomain-containing protein 4 (BRD4) is an important member of the bromodomain and extra terminal domain family that influences transcription of downstream genes by binding to acetylated histones. In the present study, we found that BRD4 was up-regulated in thyroid cancer tissues and cell lines. Inhibition of BRD4 in thyroid cancer cells by JQ1 resulted in cell cycle arrest at G0/G1 phase and enhanced (131)I uptake in vitro and suppressed tumor growth in vivo. Moreover, JQ1 treatment suppressed C-MYC but enhanced NIS expression. We further demonstrated that BRD4 was enriched in the promoter region of C-MYC, which could be markedly blocked by JQ1 treatment. In conclusion, our findings revealed that the aberrant expression of BRD4 in thyroid cancer is possibly involved in tumor progression, and JQ1 is potentially an effective chemotherapeutic agent against human thyroid cancer.

Effect of outdoor activity on myopia onset and progression in school-aged children in northeast China: the Sujiatun Eye Care Study.

BACKGROUND: Due to its high prevalence and associated sight-threatening pathologies, myopia has emerged as a major health issue in East Asia. The purpose was to test the impact on myopia development of a school-based intervention program aimed at increasing the time student spent outdoors. METHODS: A total of 3051 students of two primary (grades 1-5, aged 6-11) and two junior high schools (grades 7-8, aged 12-14) in both urban and rural Northeast China were enrolled. The intervention group (n = 1735) unlike the control group (n = 1316) was allowed two additional 20-min recess programs outside the classroom. A detailed questionnaire was administered to parents and children. Uncorrected visual acuity (UCVA) was measured using an E Standard Logarithm Vision Acuity Chart (GB11533-2011) at baseline, 6-month and 1-year intervals. A random subsample (n = 391) participated in the clinic visits and underwent cycloplegia at the beginning and after 1 year. RESULTS: The mean UCVA for the entire intervention group was significantly better than the entire control group after 1 year (P < 0.001). In the subgroup study, new onset of myopia and changes in refractive error towards myopia were direction during the study period was significantly lower in the intervention group than in the control group (3.70 % vs. 8.50 %, P = 0.048; -0.10 ± 0.65 D/year vs. -0.27 ± 0.52 D/year, P = 0.005). Changes in axial length and IOP were also significantly lower following the intervention group (0.16 ± 0.30 mm/year vs. 0.21 ± 0.21 mm/year, P = 0.034; -0.05 ± 2.78 mmHg/year vs. 0.67 ± 2.21 mmHg/year, P = 0.006). CONCLUSIONS: Increasing outdoor activities prevented myopia onset and development, as well as axial growth and elevated IOP in children. TRIAL REGISTRATION: Current controlled trials NCT02271373.

[Association between elevated light levels in classrooms and change in vision acuity among elementary and secondary students].

OBJECTIVES: To explore the association between elevated light levels in classrooms and change in vision acuity among elementary and secondary students. METHODS: A total of 4 elementary (grade 1-5) and secondary (grade 7-8) schools in urban and rural areas in Sujiatun, Shenyang, China were selected by cluster sampling as experimental schools, and lighting systems have been rebuilt to improve the ambient light levels in 56 classrooms in November 2012. The control schools were chosen for the comparable academic burden and adjacent location to experimental schools, 4 schools in all. Cluster sampling of all students in the selected schools as the subjects was carried out. A total of 2 092 students were chosen as experimental group and 1 595 students were in the control group. The luxmeter was used to measure illuminance of classrooms in two groups at baseline, and intervention for 1 month, respectively.Students in both groups were underwent 3 times for vision acuity examination by standard logarithmic visual acuity chart at baseline, intervention for 6 month and intervention for 1 year, respectively. The light levels of desk and blackboard in two groups were compared by Wilcoxon test. Multivariate analysis of covariance with repeated measures was performed to assess three vision acuity results between groups. RESULTS: After intervention, the average illuminance of desk (117.5 vs 532.5 lx, Z = -5.38, P < 0.001) and blackboard (75.6 vs 423.5 lx, Z = -5.38, P < 0.001) and uniformity of desk (Z = -4.28, P < 0.001) with new lighting were improved significantly than that with old lighting, however the uniformity of blackboard was lower than baseline significantly (0.64 vs 0.70, Z = -2.34, P = 0.019). The average scores of vision acuity in students at baseline, intervention for 6 month and intervention for 1 year were 4.87 ± 0.23, 4.84 ± 0.25 and 4.85 ± 0.23 in experimental group, and 4.88 ± 0.22, 4.84 ± 0.25 and 4.81 ± 0.27 in control group, respectively. The significant differences between groups were found and F values were 1.41, 0.13, 19.99, P values were 0.235,0.724, <0.001. At last the average vision acuity in experimental group were significantly better than that in control group either among elementary (4.90 ± 0.20) vs (4.87 ± 0.21) score, F = 13.61, P < 0.001 or secondary students (4.73 ± 0.28) vs (4.68 ± 0.32) score, F = 14.25, P < 0.001. CONCLUSIONS: Visual acuity loss could be decreased in students with elevated light levels which may slow the response to myopiagenic stimuli for eyes, therefore the ambient light levels of blackboard and desk in classroom should be improved.

Elevated light levels in schools have a protective effect on myopia.

PURPOSE: To determine whether elevated light levels in classrooms in rural areas can protect school-age children from myopia onset or myopia progression. METHODS: A total of 317 subjects from 1713 eligible students aged six to 14 in four schools located in northeast China participated in the study. Students received a comprehensive eye examination including cycloplegic refraction and ocular biometry, which included axial length (AL), anterior chamber depth (ACD), and corneal curvature (CC) measurement, and completed a questionnaire. The intervention arm included 178 students in two schools with rebuilt elevated lighting systems and the control arm included 139 students in which lighting systems were unchanged. Results for the two arms were compared with a Wilcoxon rank sum test, a chi-squared test or a t-test, as appropriate. Factors that might help explain any differences were explored with multivariate linear regression analysis. RESULTS: The median average illuminance of blackboards and desks and uniformity of desk lighting were significantly improved, however, the uniformity of blackboard lighting declined after intervention. At baseline, the mean refraction, AL, CC, ACD and myopia prevalence between the two arms were not significantly different. After 1 year, compared with the control arm the intervention arm had a lower incidence of new myopia onset (4% vs 10%; p = 0.029), a smaller decrease in refractive error among no myopic subjects (-0.25 dioptre [D] vs -0.47 D; p = 0.001), and shorter axial growth for both non-myopic (0.13 vs 0.18 mm; p = 0.023) and myopic subjects (0.20 vs 0.27 mm; p = 0.0001). Multivariate linear regression analysis showed the intervention program, lower hyperopic baseline refraction, lower father's education level, longer time sleeping and less time in screen-viewing activities were associated with less refractive shift in the direction of myopia in non-myopic children. For myopic subjects, myopia progression was significantly associated with family income only. The intervention program and older age had a protective effect on axial growth for both myopic and non-myopic subjects. The father's education level and sleep duration were significantly associated with axial growth in non-myopic children. CONCLUSIONS: Elevated light levels in classrooms have a significant effect on myopia onset, decreases in refraction, and axial growth; if the findings of lighting intervention are reproduced in future studies, the ambient light levels in schools should be improved.