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Objective: Recombinant adeno-associated virus type 8 (rAAV8) mediating transgene expression in mice was investigated using co-expressed report gene of secreted Gaussia princeps luciferase (Gluc) and non-secreted firefly luciferase(Fluc)., Methods: rAAV8-Gluc/Fluc was prepared and infected HEK293 cells to test its performance in vitro. BALB/c mice were received rAAV8-Gluc/Fluc at different doses by intravenous injection (iv) or intramuscular injection (im). Then Glue activities in blood were measured,the whole-body images for Flue activities were performed and Flue activities of tissue lysate were also detected., Results: rAAV8-Gluc/Fluc was successfully prepared and could infected HEK293 cells. The Glue was mainly detected in the culture media while the Flue was mainly detected within cells. The blood Glue activities of mice received rAAV8-Gluc/Fluc by iv or im peaked at 10-20 d post injection and persisted for at least 120d. The blood Gluc activities increased at the rAAV8-Gluc/Fluc dose-dependent manner. For mice received rAAV8 by iv, Fluc mainly expressed in liver and minor Fluc expression was also found in cardiac muscle and skeletal muscle. For mice received rAAV8-Gluc/Fluc by im, Fluc mainly expressed in local skeletal muscle and secondly in liver., Conclusion: rAAV8-Gluc/Fluc has been prepared successfully and its mediating transgene expression in mice has been investigated. This research will facilitate preclinical studies for rAAV8-mediated gene therapy.

The aim of this study was to investigate the molecular mechanisms underlying the antitumour effects of CD40L through analysing the change of genes expression profile in AGS using Affymetrix Gene Chip. Human gastric carcinoma AGS cells were first incubated with 2 μg/ml sCD40L or equal volume of medium (control) in F12 medium. RNA was isolated from AGS and were reverse transcribed, labeled with digoxigenin-11-dUTP, and then hybridized with Clontech Atlas mouse cDNA expression arrays for comparison. Performing clustering analysis, we found that 7 detected genes were down-regulated and 38 were upregulated as the sCD40L acted on AGS. To further verify the results of gene chip screening, Gene Database was searched, finding that the most significantly up-regulated genes were Gadd45a, c-Jun and Bcl-2, and the most significantly down-regulated genes were Cyclin D1, CDC6, TNFR10B, c-IAP2 and ORC5L. Based upon these findings, the signalling pathways that possibly mediate CD40-induced apoptosis are proposed.

Plectasin is a defensin-like antimicrobial peptide isolated from a fungus, the saprophytic ascomycete Pseudoplectania nigrella. Plectasin showed marked antibacterial activity in vitro against Gram-positive bacteria, especially Streptococcus pneumoniae, including strains resistant to conventional antibiotics. Plectasin could kill the sensitive strain as efficaciously as vancomycin and penicillin and without cytotoxic effects on mammalian cell viability. In order to establish a bacterium-based plectasin production system, in the present study, the coding sequence of plectasin was optimized, and then cloned into pET32a (+) vector and expressed as a thioredoxin (Trx) fusion protein in Escherichia coli. The soluble fusion protein collected from the supernatant of the cell lysate was separated by Ni(2+)-chelating affinity chromatography. The purified protein was then cleaved by Factor Xa protease to release mature plectasin. Final purification was achieved by Ni(2+)-chelating chromatography again. The recombinant plectasin exhibited the same antimicrobial activity as reported previously. This is the first study to describe the expression of plectasin in E. coli expression system, and these works might provide a significant foundation for the following production or study of plectasin, and contribute to the development and evolution of novel antimicrobial drugs in clinical applications.

A novel dual luciferase expression vector was designed and its expression characteristics were studied in vitro and in vivo. Firstly, the Gluc and Fluc genes were connected via the TaV 2A sequence by overlaping PCR, and inserted into the expression vector pAAV2neoCAG, obtaining the recombinant plasmid pAAV2neoCAG-Gluc-2A-Fluc. Then pAAV2neoCAG-Gluc-2A-Fluc was transfected into BHK21 cells and the activity of Gluc and Fluc in the supernatant and cell lysates were assayed respectively. Results showed that both Gluc and Fluc were expressed successfully. The Gluc was mainly detected in the culture media while the Fluc was mostly within cells. The activity of Gluc in the supernatant increased gradually with time while the Fluc activity in cells almost kept stable. To investigate the expression of pAAV2neoCAG-Gluc-2A-Fluc in vivo, the plasmid was hydrodynamically injected into BALB/c mice through tail vein. The Gluc activity was assayed in a small volume of blood taken by tail vein at different time points. Results showed that Gluc was expressed stably at least 7 days. Live bioluminescence imaging technology was used to compare the expression characteristics of Gluc and Fluc. Whole body imaging was seen when coelenterazine, a specific substrate for Gluc, was injected, and the imaging signals decreased rapidly within 10 minutes. Liver imaging was showed when Flue specific substrate named D-Luciferin was injected, and the imaging remained stable at least for half an hour. The dual luciferase expression vector pAAV2neoCAG-Gluc-2A-Fluc combines the advantages of the secreted report gene Gluc and the non-secreted report gene Fluc, and will provide a new tool for cell labeling and tracing.

Aim: To express human anti-keratin Fab in Pichia pastoris secretively and optimize the expression condition., Methods: Genes of kappa chain and Fd fragment of anti-keratin antibody from the plasmids p3MH/Fab were subcloned into vectors pPIC9K and pPICZalphaA respectively. After confirmed by DNA sequence analysis, the recombinant plasmids pPIC9K/L and pPICZalphaA/Fd were transducted into the genome of GS115 Pichia pastoris using two-step integrating technology. Mut(+) multiple insert transformants were screened by G418 and Zeocin. The pH value and methanol concentration was adjusted to optimize the expression condition., Results: Under the optimized expression condition, the Fab of anti-keratin antibody was efficiently secreted into the medium. Western blot assay proved that the expressed protein had specific keratin binding activity., Conclusion: The successful expression of the anti-keratin Fab in Pichia pastoris has laid a solid foundation for its further application.

Selective gene expression in cells in physiological or pathological conditions is important for the growth and development of organisms. Acetylation of histone H4 at K16 (H4K16ac) catalyzed by histone acetyltransferase 8 (KAT8) is known to promote gene transcription; however, the regulation of KAT8 transcription and the mechanism by which KAT8 acetylates H4K16ac to promote specific gene expression are unclear. Using the lepidopteran insect Helicoverpa armigera as a model, we reveal that the transcription factor FOXO promotes KAT8 expression and recruits KAT8 to the promoter region of autophagy-related gene 8 (Atg8) to increase H4 acetylation at that location, enabling Atg8 transcription under the steroid hormone 20-hydroxyecdysone (20E) regulation. H4K16ac levels are increased in the midgut during metamorphosis, which is consistent with the expression profiles of KAT8 and ATG8. Knockdown of Kat8 using RNA interference results in delayed pupation and repression of midgut autophagy and decreases H4K16ac levels. Overexpression of KAT8-GFP promotes autophagy and increases H4K16ac levels. FOXO, KAT8, and H4K16ac colocalized at the FOXO-binding region to promote Atg8 transcription under 20E regulation. Acetylated FOXO at K180 and K183 catalyzed by KAT8 promotes gene transcription for autophagy. 20E via FOXO promotes Kat8 transcription. Knockdown or overexpression of FOXO appeared to give similar results as knockdown or overexpression of KAT8. Therefore, FOXO upregulates KAT8 expression and recruits KAT8 to the promoter region of Atg8, where the KAT8 induces H4 acetylation to promote Atg8 transcription for autophagy under 20E regulation. This study reveals the mechanism that KAT8 promotes transcription of a specific gene. [ABSTRACT FROM AUTHOR]

Effective treatments for nonobstructive azoospermia (NOA), which affects 1% of all men globally, are limited by undefined pathogenic mechanisms, especially in idiopathic NOA (iNOA). Here, we tried to identify the functional ferroptosis-related genes and phenotypes involved in iNOA. Differentially expressed ferroptotic genes were identified from iNOA mRNA microarray datasets by bioinformatic analyses, and these ferroptotic genes were subsequently filtered by various algorithms. Then, receiver operating characteristic (ROC) curves were generated to evaluate the diagnostic ability of the abovementioned genes for iNOA. Generally, 11 differentially expressed ferroptotic genes were downregulated, and five genes were upregulated in iNOA samples. Four genes, including DUSP1, GPX4, HSD17B11, and SLC2A8, were technically selected and determined to be potential biomarkers for iNOA. Subsequently, similar expression levels were validated at both the RNA and protein levels in the iNOA specimens. Finally, morphologic and biochemical assays were applied to define the ferroptotic phenotypes in testes. The ferroptotic features, like shrunken mitochondria with electron-dense membranes and a reduction in cristae were observed across various cell types within iNOA patients, accompanied by the overload of ferrous ions and increased lipid peroxidation production. Our findings demonstrated that these ferroptosis genes could be involved in the underlying pathogenesis mechanisms of iNOA by regulating ferroptosis and serve as potential diagnostic biomarkers. Also, the ferroptotic phenotypes were identified in iNOA patients. [ABSTRACT FROM AUTHOR]

Soft tissue sarcomas (STS) are rare malignant tumors of mesenchymal origin, which are easy to metastasize and relapse and are a great threat to human health. In our previous study, the abnormal expression of neurofibromin 1 (NF1) is observed in tumor tissue of STS, and the NF1 gene is regulated by miRNAs. The study aimed to assess the association between NF1-related miRNA gene polymorphisms and the risk of STS. In this case–control study, the information and peripheral blood were collected from 169 patients with STS and 170 healthy controls. Six single-nucleotide polymorphisms of the NF1-related miRNAs were investigated and genotyped using a Sequenom MassARRAY® matrix-assisted laser desorption/ionization time-of-flight mass spectrometry platform. The association between the polymorphisms and the risk of STS was estimated using unconditional logistic regression analysis. There was a significant statistical difference on genotype distribution of miR-199a2 rs12139213 between the case group and the control group (p = 0.026). Comparing with individuals with wild-type AA, individuals with the AT/TT genotype had a 1.753-fold (odds ratio [OR] = 1.753, 95% confidence interval [CI] = 1.090–2.819, p = 0.021) increased risk of STS and 1.907-fold (OR = 1.907, 95% CI = 1.173–3.102, p = 0.009) increased risk of STS adjusted for age and smoking status. Individuals with the AG/GG genotype for miR24–3p rs4743988 displayed a significantly reduced risk of STS compared with individuals with homozygous mutations AA (OR = 0.605, 95% CI = 0.376–0.973, p = 0.038). Individuals carrying the AT/TT genotype for miR-199a2 rs12139213 or the AA genotype for miR24–3p rs4743988 may be susceptible to STS, which could be potential biomarkers for the diagnosis of STS. [ABSTRACT FROM AUTHOR]

Context: Oleic acid together with insulin is widely used to induce preadipocyte differentiation in humans and mammals, and is also used alone in chicken preadipocytes from abdominal adipose tissue. However, it is not clear whether oleic acid alone promotes goat primary intramuscular preadipocyte differentiation. Aims: The aim of the present study was to identify the role of oleic acid in regulating primary preadipocyte differentiation in goats. Methods: Three healthy, 7-day-old Jianzhou goats were randomly selected. After slaughter, the longissimus dorsi tissues were collected from each goat under sterile procedures and mixed equally. The primary preadipocytes were then prepared using collagenase type I digestion, and treated with 5 mg/L insulin or different concentrations of oleic acid, including 0 μM, 50 μM, 100 μM, 150 μM and 300 μM. The results were determined using microscopy and Oil Red O staining. The expression of genes related to preadipocyte differentiation were determined by real-time fluorescence quantitative polymerase chain reaction. Results: Lower concentrations of oleic acid (50 μM, 100 μM and 150 μM) did not affect the cell morphology and cell growth, whereas 300 μM oleic acid led to severe cytotoxicity compared with the control (0 μM). The treatment of oleic acid (100 μM) enhanced cellular accumulation and lipid droplets deposition significantly, which was not affected by supplementary insulin. In addition, insulin alone treatment did not alter cellular adipogenesis in goat intramuscular preadipocytes. Treatment with oleic acid significantly increased the expression of peroxisome proliferator-activated receptor gamma, CCAAT enhancer-binding protein alpha and fatty acid binding protein 4, and decreased the expression of lipoprotein esterase on Day 2 after cell differentiation, all of which decreased continually on Day 4 and Day 6. Expression of all genes increased significantly on Day 8 after oleic acid treatment in goat intramuscular preadipocytes. Conclusion: The results underscore the role of oleic acid independent of insulin in promoting intramuscular preadipocytes in goats, and probably via the control of peroxisome proliferator-activated receptor gamma and CCAAT enhancer-binding protein alpha. Implications: These data provide insight into the mechanism underlying preadipocyte differentiation. The present study found that 100 μM oleic acid, independent of insulin, enhanced the triglyceride accumulation and lipid droplet deposition in goat primary intramuscular preadipocytes, and was not affected by supplementation of insulin. Meanwhile, oleic acid significantly increased the expression of peroxisome proliferator-activated receptor gamma, CCAAT enhancer-binding protein alpha and fatty acid binding protein 4, and decreased the expression of lipoprotein esterase. Oleic acid, independent of insulin, may promote goat preadipocyte differentiation via the expression of peroxisome proliferator-activated receptor gamma and CCAAT enhancer-binding protein alpha. [ABSTRACT FROM AUTHOR]

ADGRD1 (GPR133), an adhesion G protein-coupled receptor (GPCR), has been linked to cancer. However, the prognostic value and regulatory function within non-small-cell lung cancer (NSCLC) is still unclear. This work adopted various bioinformatics methods, including publicly available databases as well as real-time PCR (RT-PCR), for detecting ADGRD1 expression level and investigating the correlation between ADGRD1 expression level and prognosis, tumor mutational burden (TMB), microsatellite instability (MSI), immune infiltrating cells, immune-related genes, and targeted regulation mechanisms in NSCLC. According to the results, ADGRD1 expression decreased within NSCLC, which might be the factor predicting prognosis of NSCLC. Meanwhile, ADGRD1 showed significant correlation with TMB and MSI, respectively, as well as immune cell infiltrating levels in lung adenocarcinoma (LUAD), which were primarily linked to macrophage M1, mast cell resting, T cell CD4 memory activated, and T cell CD4 memory resting and were associated with mast cell activated and mast cell resting in lung squamous cell carcinoma (LUSC). The most promising upstream regulation pathways of ADGRD1 were likely miR-142-5p, miR-93-5p, and miR-17-5p, which were overexpressed and associated with poor prognosis in NSCLC. ADGRD1 and immune-related genes correlated with ADGRD1 were shown to be enriched in "positive regulation of leukocyte activation," "external side of plasma membrane," "receptor ligand activity," and "cytokine-cytokine receptor interaction" pathways. ADGRD1 expression and regulation may be critical in determining NSCLC prognosis. [ABSTRACT FROM AUTHOR]

Osteosarcoma (OS) is a common primary malignancy in children and adolescents with relative high survival rate after chemotherapy. While, the toxicity of chemotherapy and personalized different response to chemotherapy makes it difficult for the selection of therapeutics and improvement of diagnosis. In this study, we conducted a combined analysis of two types of microarray datasets (gene expression and DNA methylation) from the Gene Expression Omnibus (GEO). Differential methylation sites (DMS) were identified by the IMA package and differential expression genes (DEGs) were screened out via the limma package. A total of 11,242 DMS (corresponding to 3080 genes (DMGs)) and 337 DEGs, with 40 overlaps (OS genes) between DEGs and DMGs, were identified. Enriched functions of OS genes were obtained through the Database for Annotation, Visualization and Integrated Discovery (DAVID). The OS genes were mainly enriched in the biological processes related to inflammatory/immune response and Pertussis pathways and Hematopoietic cell lineage pathways. Besides, OS-specific disease network was obtained, and found that UBS and NRF1 were regulated by multiple OS genes. Kaplan Meier analysis of OS genes identified BHMT2, DOCK2, DNALI1 and RIPK3 as significant OS survival-related genes. SEMA3A and PRAME are included in the 40 OS genes and within the top 10 most up-regulated DEGs. Their expression changes were further validated in U2OS osteosarcoma cell lines and hOB normal cell lines through quantitative PCR (qPCR) and consistent result with microarray analysis was obtained. Based on this study, some novel targets were identified for OS, which would be helpful in its early diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Due to the high mortality of lung cancer, early diagnosis followed by early effective treatment is the key to prognosis improvement, which demands to identify the biological targets. Therefore, a multistage screening analysis was used to identify biological targets of lung adenocarcinoma. Two independent datasets of DNA methylation and RNA expression microarray in lung adenocarcinoma and normal lung tissues were chosen from the Gene Expression Omnibus. The association between DNA methylation and gene expression was explored, and the prognostic value of the hub genes was also evaluated. In this study, 8533 differential methylation sites, mostly located in the CpG island region and corresponding to 2754 genes (referred as differential methylation genes), were detected from methylation microarray. Besides, we obtained 830 differential expression genes, including 570 downregulated and 260 upregulated genes, through differential expression analysis. Protein–protein interaction analysis identified CXCL12, GFR, KDR, MMP9, TEK, and TWIST as core nodes, involving in the process of tumor cell identification, cell growth, cytokine secretion, inflammatory response, and angiogenesis. Among them, MMP9 and TWIST1 were identified as more valuable biological targets for the early diagnosis and targeted therapy of lung cancer through Kaplan–Meier analysis of TCGA lung adenocarcinoma datasets. Our study should contribute to the diagnosis and treatment of lung adenocarcinoma. [ABSTRACT FROM AUTHOR]

Abstract: This study investigated the expression of heat shock protein 90 alpha (Hsp90α) in acute leukemia cells. The expression of Hsp90α was investigated in leukemia cell lines and human bone marrow mononuclear cells derived from acute leukemia patients and from healthy individuals using polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay. Compared with cells from healthy individuals, the expression of Hsp90α in the untreated patients was higher. Similarly high levels were observed in remission patients. Significantly higher expression levels were observed in all the tested cell lines, and in cells from refractory and relapsed patients. No obvious relationship was observed between the occurrence of graft versus host disease and the expression of Hsp90α. The untreated patients showing higher expression levels of Hsp90α had lower complete remission rates. During remission of untreated patients, the expression of Hsp90α decreased and reached the lowest level after transplantation, but the expression increased again before relapse. Hsp90α was highly expressed in leukemia cells. The expression level of Hsp90α was associated with leukemia prognosis. However, no obvious relationship was observed between the occurrence of graft versus host disease and the expression of Hsp90α. [Copyright &y& Elsevier]

The Vitreoscilla hemoglobin ( VHb) gene was expressed in yeast-like conidia (YLCs) of Tremella fuciformis ( T. fuciformis) to increase cell density in submerged fermentation by enhancing oxygen uptake. With the intention of doing this, an integrated expression vector containing the VHb gene and the hygromycin B phosphotransferase ( hph) gene derived from Escherichia coli ( E. coli) as the selectable marker was constructed, and then transformed into protoplasts of YLCs from T. fuciformis with restriction enzyme-mediated DNA integration (REMI). Hygromycin-resistant transformants had been generated during the transformation. Molecular evidences including PCR assay, Southern blotting, and Western blot analysis indicated the VHb gene had been integrated into the genome of transgenic T. fuciformis strains and was expressed successfully. Shake-flask fermentation and bioreactor cultivation results showed that the expression of VHb in this fungus could enhance growth of YLCs. The final cell density was higher in the culture of VHb-expressing strain than that of the wild-type strain. Moreover, these results also suggested that CaMV35S promoter was capable of driving the expression of heterologous genes in T. fuciformis. [ABSTRACT FROM AUTHOR]

Background: Cerebral hypoperfusion is a pivotal risk factor for vascular dementia (VD), for which effective therapy remains inadequate. Persistent inflammatory responses and excessive chemotaxis of microglia/macrophages in the brain may accelerate the progression of VD. Endocannabinoids are involved in neuronal protection against inflammation-induced neuronal injury. Cannabinoids acting at cannabinoid receptor 2 (CB2R) can decrease inflammation. Based on the identification of paeoniflorin (PF) as a CB2R agonist, we investigated the neuroprotective and microglia/macrophages M1 to M2 polarization promoting effects of PF in a permanent four-vessel occlusion rat model. Methods: One week after surgery, PF was intraperitoneally administered at a dose of 40 mg/kg once a day for 28 successive days. The effects of PF on memory deficit were investigated by a Morris water maze test, and the effects of PF on hippocampal neuronal damage were evaluated by light microscope and electron microscope. The mRNA and protein expression levels of key molecules related to the M1/M2 polarization of microglia/macrophages were assessed by RT-qPCR and Western blotting, respectively. Results: Administration of PF could significantly attenuate cerebral hypoperfusion-induced impairment of learning and memory and reduce the morphological and ultrastructural changes in the hippocampal CA1 region of rats. Moreover, PF promoted an M1 to M2 phenotype transition in microglia/macrophages in the hippocampus of rats. In addition to its inhibitory property against proinflammatory M1 mediator expression, such as IL-1β, IL-6, TNF-α and NO, PF dramatically up-regulated expression of anti-inflammatory cytokines IL-10 and TGF-β1. Importantly, CB2R antagonist AM630 abolished these beneficial effects produced by PF on learning, memory and hippocampus structure in rats, as well as the polarization of microglia/macrophages to the M2 phenotype. Additionally, PF treatment significantly inhibited cerebral hypoperfusion-induced mTOR/NF-κB proinflammatory pathway and enhanced PI3K/Akt anti-inflammatory pathway. Effects of PF on these signaling pathways were effectively attenuated when rats were co-treated with PF and AM630, indicating that the mTOR/NF-κB and PI3K/Akt signaling pathways were involved in the PF effects through CB2R activation. Conclusion: These findings demonstrated PF exerts its neuroprotective effect and shifts the inflammatory milieu toward resolution by modulation of microglia/macrophage polarization via CB2R activation. [ABSTRACT FROM AUTHOR]

Background: In China, Panax notoginseng has been used to treat oxidative stress-related diseases for a long time. Panax notoginseng saponins is an extract from Panax notoginseng Ledeb. Its therapeutic potential is related to antioxidant activity, but related mechanisms are still unclear. The study aims to assess the protection effects of Panax notoginseng saponins in the taurocholate-induced rat model of acute pancreatitis (AP) and explore underlying mechanisms. Methods: A rat model of severe acute pancreatitis (SAP) was established in rats induced with taurocholate. Panax notoginseng saponins was firstly administered in the treatment group via intravenous injection. After 2 h, taurocholate administration was performed. After 24 h, the expression levels of miR-181b, Beclin1, LC3-II, Akt and mTOR from pancreas tissues were measured by Western Blotting and RT-PCR. Then the expression levels of Caspase-3 and Blc-2 were determined by immunohistochemistry. Apoptosis was assessed by the TUNEL assay. Amylase and lipase in serum were determined by ELISA and pancreatic water contents in pancreatic tissue were measured. After eosin and hematoxylin staining, the histologic analysis was performed. Results: After SAP induction by taurocholate and the treatment with Panax notoginseng saponins for 24 h, we detected the up-regulated miR-181b, the reduced Bcl-2 expression, the increased activity of mTOR/Akt, the blocked Beclin1 and LC3-II expressions, and the enhanced Caspase-3 expression. Serum lipase and amylase levels were significantly decreased in the treatment group of Panax notoginseng saponins compared to the control group. Histological analysis results verified the attenuation effects of Panax notoginseng saponins on taurocholate-induced pancreas injury, apoptosis, and autophagy. Conclusion: By up-regulating the miR-181b expression level, Panax notoginseng saponins significantly reduced taurocholate-induced pancreas injury and autophagy and increased apoptosis. The significant protection effects of Panax notoginseng saponins suggested its potential in treating taurocholate induced-acute pancreatitis. [ABSTRACT FROM AUTHOR]

Plants use various secondary chemicals in their chemical defense against herbivores. While botanical insecticides are crucial for reducing the reliance on chemical pesticides, the development of plant-derived insecticides remains limited. In this study, we fed Henosepilachna vigintioctopunctata with three different host plants (Solanum nigrum, Solanum tuberosum, and Physalis floridana) and observed that feeding on P. floridana led to changes in the body size and a significantly high mortality rate. Through transcriptome analysis, it was found that the trehalose metabolism pathway of H. vigintioctopunctata changed significantly under different host feeding conditions, especially since the expression level of the trehalase gene was extremely different. We subsequently identified eight transcripts of HvTREs and analyzed their evolution and structure. Among them, significant differences are observed in the relative expression levels of HvTRE1-5 in H. vigintioctopunctata after the fourth instar and were affected by different plant diets. Compared with the natural host S. nigrum, the larvae that fed on P. floridana significantly reduced the contents of trehalose, glucose and glycogen and significantly affected the trehalase activity. Knockdown of HvTRE1-5 by RNAi increased mortality at the H. vigintioctopunctata prepupation stage, suggesting that HvTRE1-5 is important for H. vigintioctopunctata pupation. This study provides new insights into developing of green control methods for H. vigintioctopunctata and offers a valuable example for understanding the interaction between host plants and herbivorous insects. [ABSTRACT FROM AUTHOR]

• TD is an avian welfare problem cause lameness in fast-growing broiler chickens. • Therapeutic role of Celecoxib investigated in thiram-induced TD chickens. • The expression level of COX-1/2, MMP-13 and ColX genes were investigated. • Celecoxib plays a therapeutic role in the welfare problem of TD. • Celecoxib supplementation prevented chondrocytes of thiram induced TD chickens from damage. The pathogenicity of tibial dyschondroplasia (TD) is unknown; therefore, it has not been completely evacuated. In this study, we planned to find the possible therapeutic role of Celecoxib supplementation in thiram-induced TD chickens. Three hundred sixty (n = 360) broiler chickens were divided into 3 groups as control (C), thiram (T) and Celecoxib supplementary group (ST) for day 1 (8 day old), 2 (9 day old), 4 (11 day old) and 6 (13 day old). Thiram 100 mg·kg−1 was given in group (T) along with basal diet to induce TD. Supplementation of Celecoxib at 40 mg·kg−1 was given in group (ST) along with the basal diet and thiram. Clinically it was observed that broiler chickens of group T had more difficulty in locomotion than group ST. Histopathological results revealed that damaged chondrocytes increased from day 1 to 6 in group T. However, Celecoxib supplementation to thiram induced TD chickens reduced the tibial lesions significantly. Cyclooxygenase-1 (COX-1) gene was highly expressed at different stages (P < 0.01) while its protein expression was seen at a lower level. Cyclooxygenase-2 (COX-2) gene and protein expression were also changed at different stages. Expression of matrix metalloproteinase-13 (MMP-13) gene were observed high on day 1 (P < 0.01), which insignificantly changed on day 2 and 4. MMP-13 protein expression found reduced parallel to its gene expression (P < 0.01), on day 2. Type X collagen (ColX) gene expression was upregulated (P < 0.01). After adding Celecoxib in the diet, both COX-1 and COX-2 gene (P < 0.01) and their protein expression expressed similar to the control group, and to some extent observed parallel to MMP-13 gene (P < 0.01) & protein expression. In conclusion, changes in the expression of COX-1/2, MMP-13 and ColX might be related to TD occurrence. The dietary supplementation of Celecoxib plays a therapeutic role to overcome the welfare problem of TD. [ABSTRACT FROM AUTHOR]