Role of natural resources, renewable energy sources, eco-innovation and carbon taxes in carbon neutrality: Evidence from G7 economies.

Global warming has created problems for human life, and it has been increasing for a few years. All the developing and developed countries are establishing policies to attain zero carbon status. This study extends the ongoing debate on carbon emissions. It examines the effect of natural resources and RE (Biofuel and other renewable sources) on greenhouse gas (CO2 emission and PM2.5) emissions while using data over 22 years (1999-2021) from G7 countries. In addition, this study has investigated the effect of carbon taxes, financial development, and environmental policies on carbon neutrality. The cross-sectional-ARDL, the Common correlated effect means group (CCEMG), and the Augmented mean group (AMG) cutting-edge model have been employed. Quantile regression has been employed for robustness. The study results demonstrate that biofuel and other renewable energy (RE) sources, carbon taxes, environmental policy, and eco-innovation decrease greenhouse gas emissions (CO2 emissions). Meanwhile, financial development, and natural resource dependence positively impact carbon neutrality. The robustness result also verifies the findings from CS-ARDL, AMG, and CCEMG methods. The empirical findings are used to infer policy implications for G7 economies.

Multi-omics analysis and the remedial effects of Swertiamarin on hepatic injuries caused by CCl4.

Hepatic diseases pose a significant threat to community health, impacting the quality of life and longevity of millions worldwide. Despite revolutionary advancements in treatment, liver diseases remain a pressing issue, necessitating the development of more effective therapeutic approaches. Here, we conducted a comprehensive multi-omics analysis to investigate the underlying mechanism of Swertiamarin in alleviating hepatic injuries induced by CCl4 in mice. We divided 100 Kunming mice into five groups: RC (control), RM (CCl4), RD (15 mg/Kg Swertiamarin), RZ (30 mg/Kg Swertiamarin), and RG (60 mg/Kg Swertiamarin). Animals in groups RD, RZ, and RG received daily Swertiamarin via gavage, while those in groups RM, RD, RZ, and RG were treated with CCl4 solution intraperitoneally every four days, nine times in total. Our findings revealed that mice in the RM group exhibited slightly lower average weights compared to other groups, along with significantly higher liver weight (p<0.0001) and liver index (p<0.0001). Pathological analysis indicated liver damage characterized by cell degeneration, inflammatory cell infiltration, and hepatic fibrosis in the CCl4-induced group. In contrast, Swertiamarin supplementation mitigated these effects, reducing denatured cells, inflammatory cells, and collagenous fibers in the liver. Serum analysis showed elevated levels of TNF-α (p<0.001), IL-6 (p<0.05), ALT (p<0.001), AST (p<0.0001), MDA (p<0.001), and Hyp (p<0.001) in CCl4-induced animals, along with lower levels of T-AOC (p<0.001), GSH-px (p<0.0001), SOD (p<0.001), and CAT (p<0.01). Microbiome analysis revealed significant differences among groups, with pathogenic taxa such as Arthrinium and Aureobasidium, and probiotic Saccharomyces showing notable variations. Metabolomics analysis identified numerous differentially abundant metabolites, with Swertiamarin-treated animals exhibiting distinct profiles. Our findings highlight the potential of Swertiamarin ameliorating CCl4-induced liver toxicity through modulation of antioxidant capacity, inflammatory response, gut microbiota, and metabolites. These insights may inform the development of novel therapies for liver injury.

Protective effects of Abrus cantoniensis Hance against liver injury through modulation of intestinal microbiota and liver metabolites.

Abrus cantoniensis Hance (ACH) is an ancient Chinese medicine herb known for its therapeutic effects. This study investigated the potential protective effect of ACH against carbon tetrachloride (CCl4)-induced liver damage in mice. Fifty (n= 50) ICR mice were grouped into five groups. CCl4 was intraperitoneally injected into different mice groups: AM (CCl4 induced), AD (ACH-treated with 25 mg/kg), AZ (ACH-treated with 50 mg/kg), and AG (ACH-treated with100mg/kg) after every three days for a total of 31 days. The control group was denoted as AC. Additionally, groups AD, AZ, and AG received daily doses of ACH via gavage throughout the study period. According to our findings, ACH administration prominently mitigated liver pathological lesions and the increased liver index induced by CCl4 in mice (p < 0.05). Treatment with ACH resulted in a dose-dependent recovery of GSH-px, SOD, and CAT activities (p < 0.001). Moreover, the levels of TNF-α, MDA, and ALT showed significanlty decreasing trends with various doses of ACH (p < 0.001). Furthermore, 16 S rRNA gene sequencing demonstrated that ACH increased the abundance of beneficial genera of Comoclathris, Aureobasidium, and Kazachstania while decreased the presence of pathogenic genera such as Sporobolomyces and Filobasidium. Additionally, ACH treatment ameliorated the changes in liver metabolism due to CCl4 and enhanced the beneficial liver metabolites. In conclusion, ACH shows potential in protecting the liver against oxidative stress and inflammation caused by CCl4 exposure, possibly through its effects on gut microbiota and liver metabolism. Therefore, the use of ACH may offer an effective approach for alleviating CCl4-induced liver injury.

Stigmatization, panic disorder, and death anxiety among patients of Covid-19: Fourth wave of pandemic in Pakistan.

BACKGROUND: In Pakistan, the fourth wave of COVID-19 is causing an increasing number of positive cases. This fourth wave may be a risky aspect of mental health issues for COVID-19 patients. This quantitative study is designed to understand the stigmatization, and panic disorder and to explore the mediating role of death anxiety among patients of COVID-19 during the fourth wave of novel coronavirus. METHODS: The study was conducted using a correlational research design. The survey was carried out by utilizing a questionnaire with a convenient sample technique. The sample of the study was comprised of 139 patients with COVID-19. Data were collected through Stigma Scale for Chronic Illnesses (SSCI), The Panic Disorder Severity Scale (PDSS), and Death Anxiety Inventory. RESULTS: Results show that stigma is significantly positively related to panic disorder and death anxiety. Furthermore, panic disorder is also significantly positively related to death anxiety. Results also indicate that stigmatization is a significant positive predictor for death anxiety and panic disorder. Moreover, results indicate that death anxiety has a mediating role in the relationship between stigmatization and panic disorder with age and gender as covariates. CONCLUSION: This study would be helpful for people around the world to understand this threatening contagious virus so they wouldn't stigmatize infected ones. Additional research is required for the sustainable improvement of anxiety over time.

More recent insights into the breast cancer burden across BRICS-Plus: Health consequences in key nations with emerging economies using the global burden of disease study 2019.

Background: Brazil, Russia, India, China, South Africa, and 30 other Asian nations make up the BRICS-Plus, a group of developing countries that account for about half of the world's population and contribute significantly to the global illness burden. This study aimed to analyzed the epidemiological burden of female breast cancer (BC) across the BRICS-Plus from 1990 to 2019 and studied the associations with age, period, birth cohort and countries' sociodemographic index (SDI). Methods: The BC mortality and incidence estimates came from the 2019 Global Burden of Disease Study. We estimated cohort and period effects in BC outcomes between 1990 and 2019 using age-period-cohort (APC) modeling. The maximum likelihood (ML) of the APC-model Poisson with log (Y) based on the natural-spline function was used to estimate the rate ratio (RR). We used annualized rate of change (AROC) to quantify change over the previous 30 years in BC across BRICS-Plus and compare it to the global. Results: In 2019, there were about 1.98 million female BC cases (age-standardized rate of 45.86 [95% UI: 41.91, 49.76]) and 0.69 million deaths (age-standardized rate of 15.88 [95% UI: 14.66, 17.07]) around the globe. Among them, 45.4% of incident cases and 51.3% of deaths were attributed to the BRICS-Plus. China (41.1% cases and 26.5% deaths) and India (16.1% cases and 23.1% deaths) had the largest proportion of incident cases and deaths among the BRICS-Plus nations in 2019. Pakistan came in third with 5.6% cases and 8.8% deaths. Over the past three decades, from 1990 to 2019, the BRICS-Plus region's greatest AROC was seen in Lesotho (2.61%; 95%UI: 1.99-2.99). The birth cohort impacts on BC vary significantly among the BRICS-Plus nations. Overall, the risk of case-fatality rate tended to decline in all BRICS-Plus nations, notably in South Asian Association for Regional Cooperation (SAARC) and China-ASEAN Free Trade Area (China-ASEAN FTA) countries, and the drop in risk in the most recent cohort was lowest in China and the Maldives. Additionally, there was a substantial negative link between SDI and case fatality rate (r1990= -0.91, p<0.001; r2019= -0.89, p<0.001) in the BRICS-Plus in both 1990 and 2019, with the Eurasian Economic Union (EEU) nations having the highest case fatality rate. Conclusions: The BC burden varies remarkably between different BRICS-Plus regions. Although the BRICS' efforts to regulate BC succeeded, the overall improvements lagged behind those in high-income Asia-Pacific nations. Every BRICS-Plus country should strengthen specific public health approaches and policies directed at different priority groups, according to BRIC-Plus and other high-burden nations.

A Mediation Moderation Model between Self-Evaluative Emotions and Relapse Rate among Polysubstance Users: A Comparative Cross-Sectional Study.

A substantial portion of drug abuse research has concentrated on people with a single-substance-use disorder (SSUD), but many people abuse more than one drug. Studies have yet to examine how those with polysubstance-use disorder (PSUD) differ from those with an SSUD on the risk of relapse, self-evaluative emotions (e.g., shame and guilt), and personality factors (e.g., self-efficacy). Eleven rehab facilities in Lahore city, Pakistan were randomly chosen to provide a sample of 402 males with PSUD. For comparison, 410 age-matched males with SSUD were enlisted using a demographic form with eight questions, the State Shame and Guilt Scale, and the General Self-Efficacy Scale. Mediated moderation analysis was performed using Hayes' process macro. The results demonstrate that shame-proneness is positively associated with relapse rate. Guilt-proneness mediates the relationship between shame-proneness and relapse rate. Self-efficacy buffers the influence of shame-proneness on relapse rate. Although the mediation and moderation effects were found in both study groups, these effects were significantly stronger among people with PSUD than those with SSUD. To be more specific, people with PSUD reported a higher overall score on shame, guilt, and relapse rate. Additionally, people with SSUD indicated a higher score on self-efficacy than those with PSUD. The findings of this study suggest that drug rehab facilities should implement a variety of strategies to raise drug users' levels of self-efficacy, which will help to reduce their risk of relapse.

Response of cardiovascular environment to sulfonated hyaluronic acid with higher sulfur content.

Sulfonated hyaluronic acid (S-HA) has been shown to promote endothelialization in the treatment of cardiovascular diseases according to amounts of investigations. In this study, two kinds of S-HA with higher sulfur content were obtained successfully. Through a series of cell experiments, it was found that the S-HA with higher sulfur content not only possessed stronger ability of promoting the growth and migration of endothelial cells, regulating the phenotype of smooth muscle cells, but also had stronger anti-inflammatory function. Furthermore, all the S-HA molecules are very compatible with blood.

Remodeling of the microbiota improves the environmental adaptability and disease resistance in Tibetan pigs.

Introduction: The establishment of intestinal microbiota and the maintenance of its equilibrium structure plays an important role in Tibetan pigs during different growth stages. Understanding the structure and function of the intestinal microbiota at different growth stages of Tibetan pigs can provide a theoretical basis for guiding nutritional regulation and feeding management in different stages. Methods: Fecal samples were collected from the Tibetan piglets at different growth stages, and the 16S rRNA was sequenced to analyze the changes of intestinal microbiota. Results: Alpha and Beta diversity indexes showed that the diversity of the intestinal microbiota did not change during the three growth stages, and the main components of intestinal microbiota were not significantly different. At the phylum level, Firmicutes and Bacteroidetes were dominant and abundant at different growth stages and were not restricted by age. At the genus level, Streptococcus, Lactobacillus, and Bifidobacterium were the most dominant in the TP10d and TP40d groups, Streptococcus was the most dominant in the TP100d group, followed by Treponema_2 and Lactobacillus. Fusobacteria, Gluconobacter, and Synergistetes were found to be specific genera of 10-day-old Tibetan piglets by LEfSe combined with LDA score. The change of diet made Tenericutes and Epsilonbacteraeota, which are closely related to digestive fiber, become specific bacteria at the age of 40 days. With the consumption of oxygen in the intestine, obligate anaerobes, such as Verrucomicrobia, Fibrobacter, and Planctomycetes, were the characteristic genera of 100 days. KEGG function prediction analysis showed that the intestinal microbiota function of Tibetan pigs changed dynamically with the growth and development of Tibetan piglets. Discussion: In conclusion, the structure and composition of the intestinal microbiota of Tibetan pigs are significantly different at different growth and development stages, which plays an important role in their immune performance.

Shame and guilt activations: Associations and sociodemographic differences among ploysubstance abusers.

Objectives: Shame and guilt in polysubstance abusers are still understudied despite their significance in substance use disorders (SUD). The goal of the current study is to develop a better understanding of how shame and guilt interact among polysubstance abusers who are receiving residential treatment. Methods: The sample of two hundred four males with SUD admitted to five rehabilitation centers from two cities in Pakistan participated in this study. For comparison, 215 age-matched healthy individuals were recruited (control). All participants reported their scores on the state shame and guilt scale (SSGS) and demographic form. A cross-sectional study design was adopted. Results: The group with SUD reported greater activations on SGSS (r = 0.79, p < 0.001) as compared to healthy (control) individuals (r = 0.48, p < 0.001). Further, multivariate analysis indicated that people with SUD who were of older age, unemployed, living in a nuclear family system, with a higher level of education, and low income, experienced higher levels of shame and guilt. Multinomial logistic regression analysis revealed that people with SUD in the age group 41-60 years (OR 5.2, 95%CI 2.4-6.8), unemployed (OR 4.4, 95%CI 3.2-4.7), nuclear family system (OR 5.9, 95%CI 4.5-6.4) and low monthly income group (OR 5.4, 95%CI 3.5-5.8) had a significantly high risk of shame and guilt than the control group. Conclusion: Findings of the current study indicate an association between shame and guilt activation and SUD. These results suggest that polysubstance users may benefit from therapeutic interventions to avoid a generalization of shame and guilt toward their substance use. Reducing shame and guilt should be considered a priority in treating adults with multiple SUD.

Hepatotoxicity and the role of the gut-liver axis in dogs after oral administration of zinc oxide nanoparticles.

Zinc oxide nanoparticles (ZnO NPs) have been used in many fields, and people are concerned about its effects on health. The present study reported the changes in liver metabolites and intestinal microbiota induced by overused ZnO NPs in dogs and explored the related mechanisms of liver injury induced by ZnO NPs. The results showed that overused ZnO NPs promote zinc accumulation in the liver and increase liver coefficient and serum liver-related indexes. In addition, the overuse of ZnO NPs increase the reactive oxygen species levels, affecting the hepatocyte antioxidant capacity and mitochondrial function. Results showed that ZnO NPs significantly inhibited the hepatocyte apoptosis via the Cytc pathway and promoted the autophagy via activating the mTOR/ATG5 pathway. Metabolic analysis of liver tissue showed that 81 metabolites changed overall and mainly affected the glycerophospholipid metabolism. ZnO NPs can significantly change the richness and diversity of the intestinal bacteria in dogs, increasing the abundance of Firmicutes and Actinobacteria while reducing the bacterial abundance of Proteobacteria. In conclusion, the results suggest that overexposure to ZnO NPs can lead to the disruption of intestinal microbiome and liver metabolites in dogs, which ultimately leads to liver damage.

Meta-analysis on the prevalence of bovine hydatid disease in China from 2000 to 2021.

Cystic Echinococcosis (hydatidosis) is caused by the larval stage of Echinococcus spp. It is an animal-borne zoonotic parasitic disease with local epidemic and natural foci, which is very common in northwest China. In recent years, a considerable attention has been paid to the epidemic investigation of hydatid disease in humans and sheep, but there are few large-scale epidemic investigation and data analysis of bovine hydatid disease. We systematically reviewed and analyzed the prevalence of bovine hydatid disease (2000-2021) in China for the first time. Several databases including CNKI, Wanfang, VIP Chinese periodical database, Baidu Library, PubMed and ScienceDirect were used to search 57 articles and 72 sets of valid data about bovine hydatid disease in China from 2000 to 2021. We used the random effect model in META package of R software, and PAS for rate conversion. The subgroup analysis and univariate meta regression analysis were used to reveal the factors leading to the heterogeneity of the study. The total prevalence rate of bovine hydatid disease in China from 2000 to 2021 is estimated to be 17.27% (10898/63113). According to the analysis of sampling years, the lowest positive rate since 2016 is 7.54% (1503/19929). The highest prevalence rate of bovine hydatid disease is 53.93% (4340/8048). The infection rate of bovine liver accounted for the highest proportion of the total infections, 45.2% (2040/4507). We also assessed the effects of different geographical and climatic factors on the prevalence of bovine hydatid disease. The results showed that the prevalence rate of hydatid disease was higher in cold and humid areas. Although the infection rate of bovine hydatid disease has declined in recent years, it is still necessary to carry out long-term surveillance and control of hydatid disease, cut off the infection route and reduce the risk of infection in high-risk areas.

Breast Cancer Mortality Trends and Predictions to 2030 and Its Attributable Risk Factors in East and South Asian Countries.

Background: Amidst the rising breast cancer burden in Asia, we aim to predict the future mortality risk due to breast cancer and identify the risk-attributable deaths for breast cancer among East and South Asian countries. Methods: We used country-level data to predict the trends in the next decade relating to female breast cancer mortality by employing data from 1990 to 2019 from the Global Burden of Disease 2019 study. We used the stochastic mortality modeling and prediction techniques to forecast the age-specific and risk-attributable breast cancer mortality trends at the regional and national levels of East and South Asia. Results: The number of deaths caused by the breast cancer is predicted to increase in East and South Asian countries in the next decade (2020-2030). Age-standardized death rate (ASDR) of breast cancer is predicted to increase by 7.0% from 9.20/100,000 (95% CI: 6.04-12.12) in 1990 to 9.88/100,000 (95% CI: 7.12-11.4) in 2030 in East Asia, and about 35% increase from 13.4/100,000 (95% CI: 9.21-16.02) in 1990 to 18.1/100,000 (95% CI: 13.23-21.10) in 2030 in South Asia. At the national level, the highest percent change in ASDR between 1990 and 2030 was reported in Pakistan (a 62% increase) and Nepal (a 47% increase). The highest percent change in breast cancer mortality between 2020 and 2030 for females of age group 80-84 years was observed in Pakistan [21.6, (95% CI, 20.6-94.7)], followed by Afghanistan [13.3 (4.0-80.8)], and Nepal [36.6 (11.1-125.7)] as compared to the other countries. In the females of aged 50-80 years, the predicted death rates were associated with high body mass index, high-fasting plasma glucose, and diet high in red meat, across the majority of countries under study. Furthermore, reductions in percent change in mortality rates occurred in several countries with increases in sociodemographic index (SDI), notably across high SDI countries. Conclusion: Breast cancer mortality risk varies substantially across East and South Asian countries with higher mortality risk in low/middle SDI countries. Early detection using screening, awareness among females and health workers, and cost-effective and timely treatment of patients with breast cancer is vital in stemming the tide of breast cancer in the next decade.

Challenges of fluoride pollution in environment: Mechanisms and pathological significance of toxicity - A review.

Fluoride is an important trace element in the living body. A suitable amount of fluoride has a beneficial effect on the body, but disproportionate fluoride entering the body will affect various organs and systems, especially the liver, kidneys, nervous system, endocrine system, reproductive system, bone, and intestinal system. In recent years, with the rapid development of agriculture and industry, fluoride pollution has become one of the important factors of environmental pollution, and fluoride pollution in any form is becoming a serious problem. Although countries around the world have made great breakthroughs in controlling fluoride pollution, however fluorosis still exists. A large amount of fluoride accumulated in animals will not only produce the toxic effects, but it also causes cell damage and affect the normal physiological activities of the body. There is no systematic description of the damage mechanism of fluoride. Therefore, the study on the toxicity mechanism of fluoride is still in progress. This review summarizes the existing information of several molecular mechanisms of the fluoride toxicity comprehensively, aiming to clarify the toxic mechanism of fluoride on various body systems. We have also summerized the pathological changes of those organ systems after fluoride poisoning in order to provide some ideas and solutions to the reader for the prevention and control of modern fluoride pollution.

Thiram exposure in environment: A critical review on cytotoxicity.

Thiram is used in large quantities in agriculture and may contaminate the environment by improper handling or storage in chemical plants and warehouses. A review of the literature has shown that thiram can affect different organs in animals and its toxic mechanisms can be elucidated in more detail at molecular level. We have summarized several impacts of thiram on animals: the effects of the perspectives of oxidative stress, mitochondrial damage, autophagy, apoptosis, and the IHH/PTHrP pathway on regulating abnormal skeletal development in particular tibial dyschondroplasia and kyphosis; angiogenesis inhibition was investigated from the perspective of angiogenesis factor inhibition, PI3K/AKT signaling pathway and CD147; the inhibition effect of thiram on fibroblasts and erythrocytes via the perspective of oxidative stress, mitochondrial damage and inhibition of growth factors in animal skin fibroblasts and erythrocytes; studied fertilized egg size, reduced fertility, neurodegeneration, and immune damage from the perspectives of CYP51 inhibition and dopamine-b-hydroxylase inhibition in the reproductive system, vitamin D deficiency in the nervous system, and inflammatory damage in the immune system; embryonic dysplasia in terms of thyroid hormone repression in animal embryonic development and repression of the SOX9a transcription factor. The elucidation of the mechanisms of toxicity of thiram on various organs of animals at molecular level will enable a more detailed understanding of the mechanisms of toxicity of thiram in animals and will facilitate the exploration of the treatment of thiram poisoning at molecular level.

Coexistence of virulence and ß-lactamase genes in avian pathogenic Escherichia coli.

Emergence of multidrug resistance in E. coli and advent of newer strains is becoming serious concern which requires keen observations. This study was designed to find the ciprofloxacin resistant E. coli isolates co-existed with multi-drug resistance along with ß-lactamase production from poultry source, and finally the genome sequencing of these strains to explore genetic variations. Study constituted on isolation of n = 225 E. coli from broiler farms of central China which were further subjected to identification of resistance against ciprofloxacin followed by antibiogram of n = 26 antibiotics and identification of ß-lactamase production. Whole genome resequencing was performed using Illumina HiSeq 4000 system. PCR results revealed predominant ß-lactamase genes i.e.CTX-M, CTX-M-1, CTX-M3, TEM-1 and OXA. Furthermore, the MDR isolates were containing most of the tested virulence genes. The most prevalent virulence genes were pap-C, fim-C, fim-H, iuc-D, irp-2, tra-T, iro-N and iut-A. The single nucleotide polymorphisms (SNPs) loci mentioned in this data give valuable genetic markers to growing high-throughput techniques for fine-determination of genotyping of MDR and virulent isolates. Characterization of SNPs on functional basis shed new bits of knowledge on the evolution, disease transmission and pathogenesis of MDR E. coli isolates. In conclusion, these findings provide evidence that most of poultry E. coli are MDR, ß-lactamase producers, and virulent which could be a zoonotic threat to the humans. The whole genome resequencing data provide higher resolution of resistance and virulence characteristics in E. coli which can further be used for the development of prevention and treatment strategies.

Effect of paraquat on cytotoxicity involved in oxidative stress and inflammatory reaction: A review of mechanisms and ecological implications.

Paraquat (PQ) is a cheap and an effective herbicide, which is widely being used worldwide to remove weeds in cultivated crop fields. However, it can cause soil and water pollution, and pose serious harm to the environment and organisms. Several countries have started to limit or prohibit the use of PQ because of the increasing number of human deaths. Its toxicity can damage the organisms with a multi-target mechanism, which has not been fully understood yet. That is why it is hard to treat as well. The current research on PQ focuses on its targeted organ, the lungs, in which PQ mostly trigger pulmonary fibrosis. While there is a lack of systematic research, there are few studies published discussing its toxic effects at systematic level. This review summarizes the major damages caused by PQ in different organisms and partial mechanisms by which it causes these damages. For this purpose, we consulted several research articles that studied the toxicity of PQ in various tissues. We also listed some drugs that can be used to alleviate the toxicity of PQ. However, at present, the effectiveness of these drugs is still being explored in animal experiments and the study of their mechanism will also help in understanding the poisoning mechanism of PQ, which will ultimately lead to effective treatment in future.

Effect of total flavonoids of Rhizoma Drynariae in thiram induced cytotoxicity of chondrocyte via BMP-2/Runx2 and IHH/PTHrP expressions.

Tibial Dyschondroplasia (TD) is a prevailing skeletal disorder that mainly affects rapidly growing avian species. It results in reduced bone strength, lameness and an increase risk of fragility fractures. Total flavonoids of Rhizoma drynariae (TFRD) have been used as an effective treatment of different bone diseases in humans. The current in vitro study was conducted to explore the therapeutic effect of TFRD on thiram-induced cytotoxicity in avian growth plate cells via bone morphogenetic protein-2/runt related transcription factor-2 (BMP-2/Runx2) and Indian hedgehog/Parathyroid hormone-related peptide (IHH/PTHrP) expressions. Chondrocytes were isolated, cultured and refined from chicken's tibial growth plates in a special medium. Then chondrocytes were treated with sublethal thiram having less concentration (2.5 µg/mL) to induce cytotoxicity of chondrocyte, and then treated with providential doses (100 µg/mL) of TFRD. Thiram caused distorted morphology of chondrocytes, nuclei appeared disintegration or lysed along with decreased expressions of BMP-2/Runx2 and IHH/PTHrP. TFRD administration not only enhanced the viability of chondrocytes by itself, but also well restored the damage caused by thiram on growth plate chondrocytes by significantly up-regulating the expressions of BMP-2/Runx2 and IHH/PTHrP. Therefore, this study provides a novel insight into the further treatment of TD and other skeletal ailments and lays the foundation for prevention and treatment.

Effect of Anacardic Acid against Thiram Induced Tibial Dyschondroplasia in Chickens via Regulation of Wnt4 Expression.

Tibial dyschondroplasia (TD) is a tibia bone problem in broilers. Anacardic acid (AA) is a traditional Chinese medicine, which is commonly used to treat arthritis in human. The purpose of the present study is to investigate the effect of AA against TD. A total of 300 day-old poultry birds were equally divided and distributed into three different groups: Control, TD and AA groups. The results showed that the feed conversion ratio was significantly lower in the TD group than control chickens. The tibia bone parameters including weight, length and width were of low quality in TD chickens, while the width of the tibial growth plate was enlarged remarkably. Whereas, in the AA treatment group, the tibia bone parameters showed improvement and tend to return to normal. The antioxidant parameters level of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), total and antioxidant capacity (T-AOC) was significantly decreased, while malondialdehyde (MDA) level was increased significantly in TD affected chickens. AA administration restored the antioxidant parameters significantly. The gene expression revealed a decrease in Wnt4 expression in TD chickens as compared to control chickens, while AA treatment up-regulated the Wnt4 expression. The present study demonstrates that the AA plays an important role to prevent the lameness and restore the size of tibial growth plate of chickens by regulating the expression of Wnt4.

Identification of differentially expressed MiRNAs profile in a thiram-induced tibial dyschondroplasia.

Tetramethyl thiuram disulfide (thiram) is a dithiocarbamate, which is widely used on seeds and storing food grains. The incorporation of thiram into the food chain could be a risk for both human beings and animals. Thiram-contaminated feed has been considered a common cause of tibial dyschondrolplasia (TD) in many avian species. The molecular mechanism of action of thiram on TD involving microRNA (miRNA) is not fully understood. For this purpose, the morbidity and pathologic changes were evaluated to understand the TD, and high-throughput RNA sequencing (RNA-Seq) was performed to explore the differentially expressed miRNAs (DEGs). RT-qPCR was used to confirm the validity as compared with sequencing data. The results showed that the marked alterations in the growth plate of the TD chickens were noticeable, with shrinking cells and irregular chondrocyte columns as compared with control group. In this study, we identified total 375 (p < 0.1), 340 (p < 0.05) and 266 (p < 0.01) significant DEGs between the TD and control groups. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs showed that the target miRNAs were significantly enriched in different treatment groups, such as apoptosis, mRNA surveillance pathway, mitophagy-animal, etc. This study provides theoretical basis for in-depth understanding the pathogenesis of thiram-induced TD and explore the new insights towards the proposed molecular mechanism of specific miRNA as biomarkers for effective gene diagnosis and treatment of TD in broilers.