Identification of m6A-associated genes as prognostic and immune-associated biomarkers in Wilms tumor.

OBJECTIVES: Wilms tumor (WT) is a common renal malignant tumor in children. We aimed to investigate the potential prognostic value of m6A-related genes and their relationship to the immune microenvironment in WT. METHODS: RNA-seq data and clinical information from 121 WT and 6 normal samples were obtained from the University of California Santa Cruz Xena database. We used various bioinformatics analysis tools to analyze these data and verify the expression level of m6A-related genes by experiments. RESULTS: Four m6A-related genes were successfully screened, including ADGRG2, CPD, CTHRC1, and LRTM2. Kaplan-Meier survival curves showed that the four genes were closely related to the prognosis of WT, which was also confirmed by receiver operator characteristic curves. Subsequently, in the immune microenvironment of WT, we discovered that Th1_cells were positively correlated with ADGRG2, CCR was negatively correlated with CPD, CCR was positively correlated with CTHRC1, APC_co_stimulation, CCR, Macrophages, inflammation-promoting cells, Treg, and Type_II_IFN_Reponse were negatively correlated with LRTM2. Finally, qRT-PCR showed that expression levels of the four genes were upregulated in the nephroblastoma cell lines (G-401, SK-NEP-1, and WT-CLS1) compared with the human embryonic kidney cell lines (293T). CONCLUSIONS: Taken together, our study first time screened the m6A-related genes and revealed that ADGRG2, CPD, CTHRC1, and LRTM2 are the prognostic and immune-associated biomarkers in WT.

Effect of Epichloë Endophyte on the Growth and Carbon Allocation of Its Host Plant Stipa purpurea under Hemiparasitic Root Stress.

Epichloë endophytes not only affect the growth and resistance of their host plants but also confer nutrient benefits to parasitized hosts. In this study, we used Pedicularis kansuensis to parasitize Stipa purpurea, both with and without endophytic fungi, and to establish a parasitic system. In this study, endophytic fungal infection was found to increase the dry weight of the leaf, stem, and leaf sheath, as well as the plant height, root length, tiller number, aboveground biomass, and underground biomass of S. purpurea under root hemiparasitic stress. Meanwhile, the 13C allocation of the leaf sheaths and roots of S. purpurea increased as the density of P. kansuensis increased, while the 13C allocation of the leaf sheaths and roots of E+ S. purpurea was lower than that of E- S. purpurea. The 13C allocation of the stem, leaf sheath, and root of E+ S. purpurea was higher than that of its E- counterpart. Furthermore, the content of photosynthetic 13C and the 13C partition rate of the stems, leaves, roots, and entire plant of S. purpurea and P. kansuensis transferred from S. purpurea increased as the density of P. kansuensis increased. These results will generate new insights into the potential role of symbiotic microorganisms in regulating the interaction between root hemiparasites and their hosts.

Changes in bud bank and their correlation with plant community composition in degraded alpine meadows.

Bud banks are considered a crucial factor in regulating the species composition of grassland communities and maintaining the ecological function of alpine grasslands. However, few studies have paid attention to the dynamic changes of bud banks from undisturbed to severely degraded alpine meadows. Therefore, this study examined the correlations between plant diversity and bud bank traits at different stages of alpine meadows degradation. Grass biomasses and plant diversity were found to be highest in moderately degraded meadows, and sedge biomasses were highest in lightly degraded meadows. Lack of disturbance and moderate disturbance by herbivores increased the bud bank density of alpine meadows. Consistent with the changes in bud bank density, bud bank diversity was highest in undisturbed meadows. The structural equation model indicated that the densities of rhizome and the densities and diversities of tiller buds play crucial roles in facilitating the greater diversity of the plant community. Our findings suggest that the diversities and densities of rhizome and tiller buds in the degradation stages are synchronized with changes in plant diversity, and in the regenerative ability of bud banks, which largely determine the outcome of restoration in degraded meadows. These findings could provide a frame of reference for effectively restoring degraded alpine regions by regenerating bud banks. The potential driving force and renewal capacity of bud banks should be taken into account in restoring the Qinghai-Tibet Plateau's degraded meadow.

Overexpression of ST7-AS1 Enhances Apoptosis and Inhibits Proliferation of Papillary Thyroid Carcinoma Cells Via microRNA-181b-5p-Dependent Inhibition Tripartite Motif Containing 3.

Long non-coding RNAs (lncRNAs) are of great significance in the pathogenesis and progression of papillary thyroid carcinoma (PTC). LncRNA tumorigenicity 7 antisense RNA 1 (ST7-AS1) is a newly identified lncRNA serving as an oncogene or tumor suppressor in different tumors; however, the role of ST7-AS1 in PTC remains completely unknown. In this study, ST7-AS1 was mainly distributed in the cytoplasm of PTC cells and presented reduced expression in THCA tumors and PTC cell lines. Functional experiments revealed that overexpressed ST7-AS1 inhibited the viability and proliferation of PTC cells, whereas accelerated the apoptosis of PTC cells. The expression of miR-181b-5p was upregulated and it bound with ST7-AS1 in PTC cells. Moreover, TRIM3 exhibited downregulated expression level in PTC cells and ST7-AS1 elevated TRIM3 expression via harboring miR-181b-5p. Rescue experiments illuminated that knockdown of TRIM3 reversed ST7-AS1 overexpression-induced promotion on PTC cell proliferation and suppression on PTC cell apoptosis. Overall, overexpression of ST7-AS1 enhances apoptosis and represses proliferation of PTC cells via targeting the miR-181b-5p/TRIM3 axis, which may help broaden the horizon and establish the foundation to develop therapeutic strategies for PTC in the future.

Hepatic thyroid hormone signalling modulates glucose homeostasis through the regulation of GLP-1 production via bile acid-mediated FXR antagonism.

Thyroid hormones (TH) regulate systemic glucose metabolism through incompletely understood mechanisms. Here, we show that improved glucose metabolism in hypothyroid mice after T3 treatment is accompanied with increased glucagon-like peptide-1 (GLP-1) production and insulin secretion, while co-treatment with a GLP-1 receptor antagonist attenuates the effects of T3 on insulin and glucose levels. By using mice lacking hepatic TH receptor ß (TRß) and a liver-specific TRß-selective agonist, we demonstrate that TRß-mediated hepatic TH signalling is required for both the regulation of GLP-1 production and the insulinotropic and glucose-lowering effects of T3. Moreover, administration of a liver-targeted TRß-selective agonist increases GLP-1 and insulin levels and alleviates hyperglycemia in diet-induced obesity. Mechanistically, T3 suppresses Cyp8b1 expression, resulting in increased the levels of Farnesoid X receptor (FXR)-antagonistic bile acids, thereby potentiating GLP-1 production and insulin secretion by repressing intestinal FXR signalling. T3 correlates with both plasma GLP-1 and fecal FXR-antagonistic bile acid levels in people with normal thyroid function. Thus, our study reveals a role for hepatic TH signalling in glucose homeostasis through the regulation of GLP-1 production via bile acid-mediated FXR antagonism.

Triiodothyronine (T3) promotes brown fat hyperplasia via thyroid hormone receptor α mediated adipocyte progenitor cell proliferation.

The thyroid hormone (TH)-controlled recruitment process of brown adipose tissue (BAT) is not fully understood. Here, we show that long-term treatment of T3, the active form of TH, increases the recruitment of thermogenic capacity in interscapular BAT of male mice through hyperplasia by promoting the TH receptor α-mediated adipocyte progenitor cell proliferation. Our single-cell analysis reveals the heterogeneous nature and hierarchical trajectory within adipocyte progenitor cells of interscapular BAT. Further analyses suggest that T3 facilitates cell state transition from a more stem-like state towards a more committed adipogenic state and promotes cell cycle progression towards a mitotic state in adipocyte progenitor cells, through mechanisms involving the action of Myc on glycolysis. Our findings elucidate the mechanisms underlying the TH action in adipocyte progenitors residing in BAT and provide a framework for better understanding of the TH effects on hyperplastic growth and adaptive thermogenesis in BAT depot at a single-cell level.

Depletion of hepatic forkhead box O1 does not affect cholelithiasis in male and female mice.

Cholelithiasis is one of the most prevalent gastroenterological diseases and is characterized by the formation of gallstones in the gallbladder. Both clinical and preclinical data indicate that obesity, along with comorbidity insulin resistance, is a predisposing factor for cholelithiasis. Forkhead box O1 (FoxO1) is a key transcription factor that integrates insulin signaling with hepatic metabolism and becomes deregulated in the insulin-resistant liver, contributing to dyslipidemia in obesity. To gain mechanistic insights into how insulin resistance is linked to cholelithiasis, here we determined FoxO1's role in bile acid homeostasis and its contribution to cholelithiasis. We hypothesized that hepatic FoxO1 deregulation links insulin resistance to impaired bile acid metabolism and cholelithiasis. To address this hypothesis, we used the FoxO1LoxP/LoxP-Albumin-Cre system to generate liver-specific FoxO1-knockout mice. FoxO1-knockout mice and age- and sex-matched WT littermates were fed a lithogenic diet, and bile acid metabolism and gallstone formation were assessed in these animals. We showed that FoxO1 affected bile acid homeostasis by regulating hepatic expression of key enzymes in bile acid synthesis and in biliary cholesterol and phospholipid secretion. Furthermore, FoxO1 inhibited hepatic expression of the bile acid receptor farnesoid X receptor and thereby counteracted hepatic farnesoid X receptor signaling. Nonetheless, hepatic FoxO1 depletion neither affected the onset of gallstone disease nor impacted the disease progression, as FoxO1-knockout and control mice of both sexes had similar gallstone weights and incidence rates. These results argue against the notion that FoxO1 is a link between insulin resistance and cholelithiasis.

The mechanism of freeze-thawing induced accumulation of γ-aminobutyric acid in germinated soybean.

BACKGROUND: γ-Aminobutyric acid (GABA) is a non-protein amino acid with several functions in the human body. Although freeze-thawing could effectively accumulate GABA in soybean sprouts, the mechanism has not been revealed. The mechanism by which freeze-thawing enhances GABA accumulation in germinated soybean was revealed by evaluating GABA content, the activity of related synthesis enzymes, and the microstructure of the tissues and cells of sprouts. The germinated soybeans were treated at different temperatures (from -196 °C to 25 °C) for 12 h and then thawed at 25 °C for 6 h. RESULTS: The results showed that GABA content in frozen soybean sprouts did not change significantly before thawing. After thawing, the GABA content of sprouts increased by 83.9% and 82.9% when treated by liquid nitrogen flash freeze at - 80 °C for 12 h compared with the control (4 °C treatment for 12 h). The results indicated that GABA formation mainly occurred during thawing. However, glutamate decarboxylase (GAD), diamine oxidase (DAO), and aminoaldehyde dehydrogenase (AMADH) activity decreased during thawing. Based on the malonaldehyde (MDA) content and microstructure of sprouts, it was suggested that freezing at lower temperatures (< -20 °C) maintained the integrity of the cell structure, while the tissues and cell membranes were broken during thawing. CONCLUSION: These results could provide evidence for the hypothesis that GABA formation resulted from full contact between enzymes and substrates during thawing, rather than the contribution of higher enzyme activity. © 2019 Society of Chemical Industry.

AMADH inhibitor optimization and its effects on GABA accumulation in soybean sprouts under NaCl-CaCl2 treatment.

ABSTRACT: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) was the suitable inhibitor for aminoaldehyde dehydrogenase (AMADH) compared with N-ethylmaleimide (NEE) and iodoacetamide (IAM). EDC exhibited the most obvious inhibition effect on AMADH activity, while its inhibition on glutamate decarboxylase (GAD) was insignificant. Compared with the control, AMADH activity reduced by 70.4% with 0.5 mM EDC, and γ-aminobutyric acid (GABA) content declined by 44.3% in soybean sprouts at 4 days of germination. AMADH activity reduced by 80.62, 67.61 and 72.02% in the 4-day sprouts with 1 mM EDC under NaCl, CaCl2 and NaCl + CaCl2 treatment, respectively, and GABA content decreased by 43.56, 38.84 and 35.53%, respectively. EDC is a proper inhibitor for AMADH and it could be used to quantify the contribution of polyamine degradation pathway on GABA formation. In soybean sprouts, the presence of CaCl2 under NaCl stress decreased the contribution of polyamine degradation pathway on GABA accumulation.

Obesity-induced upregulation of miR-361-5p promotes hepatosteatosis through targeting Sirt1.

OBJECTIVE: Obesity is associated with an increased risk of many metabolic disorders, including non-alcoholic fatty liver disease (NAFLD). However, the underlying mechanisms remain poorly understood. Recent studies have demonstrated that MicroRNA-mediated gene silencing plays an important role in hepatic triglyceride (TG) metabolism. In the present study, we aimed to investigate the pathological function of miR-361-5p in the development of NAFLD. METHODS: Expression levels of miR-361-5p was determined by quantitative real-time PCR in livers of obese mice and NAFLD patients. Liver tissues from mice with miR-361-5p overexpression or inhibition were collected and analyzed by TG contents, gene expression profile. RESULTS: Expression of miR-361-5p was increased in the livers of two obese mouse models and NAFLD subjects. Overexpression of miR-361-5p in C57BL/6 mice led to hepatosteatosis, whereas inhibition of miR-361-5p expression in db/db mice improved TG accumulation and insulin sensitivity. Mechanistically, we identified Sirt1 as a direct target gene of miR-361-5p and re-introduction of Sirt1 largely abolished the metabolic action of miR-361-5p. CONCLUSIONS: Our results demonstrated the role of miR-361-5p in the regulation of hepatic TG homeostasis, which may provide potential therapeutic target for hepatosteatosis.

Long non-coding RNA MEG3 regulates proliferation, apoptosis, and autophagy and is associated with prognosis in glioma.

PURPOSE: Accumulating evidence indicates that dysregulated long noncoding RNAs (lncRNAs) play critical roles in tumorigenesis and cancer progression. LncRNA-maternally expressed gene 3 (MEG3) has been shown to be involved in the initiation and development of several cancers, including glioma. However, the clinical prognostic value of MEG3 in glioma has not yet been fully elucidated. METHODS: The expression levels of MEG3 were detected in 79 glioma tissues and adjacent normal brain tissues, as well as, glioma cells and normal human astrocytes by qRT-PCR. Kaplan-Meier and Cox regression methods were utilized for the survival analysis. MTT assay, flow cytometry, and immunofluorescence assay were carried out to detect the impact of MEG3 on glioma cell proliferation, apoptosis, and autophagy. RESULT: The current results showed that MEG3 expression was significantly downregulated in glioma tissues and cell line and negatively correlated with WHO grade in glioma patients. Low MEG3 expression was significantly associated with the advanced WHO grade, low Karnofsky performance score (KPS), IDH wild-type, and tumor recurrence. Patients displaying a low expression of MEG3 contributed to poor overall survival. The downregulated level of MEG3, advanced WHO grade, low KPS, IDH wild-type, and tumor recurrence were independent poor prognostic indicators in glioma patients. The in vitro experiments demonstrated that the MEG3 overexpression remarkably suppressed the proliferation while facilitating apoptosis and autophagy in glioma cells. CONCLUSIONS: These findings indicated a critical role of MEG3 in glioma cell proliferation, apoptosis, and autophagy. Also, the gene was found to be significantly associated with the prognosis in glioma patients. Thus, it might provide a new target for predicting prognosis and therapeutic intervention in glioma.

Association between SKIV2L polymorphism rs429608 and age-related macular degeneration: A meta-analysis.

PURPOSE: This study was conducted to comprehensively evaluate the potential association of SKIV2L polymorphism rs429608 with age-related macular degeneration (AMD) through a meta-analysis. METHODS: We performed a literature search in EMBASE, PubMed, Web of Science, and the Chinese Biomedical Database for AMD genetic studies published before August 30, 2015. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated for single-nucleotide polymorphisms (SNPs) using fixed-effect models or random effect models according to between-study heterogeneity. Publication bias analyses were conducted using Egger's test. RESULTS: A total of five studies from published articles were included, and a total number of 2789 AMD cases and 3451 healthy controls were tested in this meta-analysis. The results demonstrated that SKIV2L rs429608 is associated with AMD under allelic model (A vs. G; OR = 0.52, 95% CI 0.44-0.62, p < 0.001), heterozygous model (AG vs. GG; OR = 0.51; 95%CI, 0.38-0.68; p < 0.001; PQ = 0.48; I2 = 0) and dominant model (AA+AG vs. GG; OR = 0.49; 95%CI 0.37-0.65; p < 0.001; PQ = 0.44; I2 = 0), but not under other genetic models. CONCLUSIONS: This meta-analysis showed that SKIV2L rs429608 was statistically associated with AMD and it might exert a protective effect on AMD. Further investigations are needed to validate the association and confirm the role of SKIV2L in AMD.

Effects of dexamethasone and HA1077 on actin cytoskeleton and ß-catenin in cultured human trabecular meshwork cells.

AIM: To investigate the effects of dexamethasone (DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine (HA1077) on actin cytoskeleton and ß-catenin in cultured human trabecular meshwork (HTM) cells. METHODS: The HTM cells were separated from human eyeball and cultured in vitro. They were divided into control group, DEX (1×10-6 mol/L) group, HA1077 (3×10-5 mol/L) group, and DEX (1×10-6 mol/L) and HA1077 (3×10-5 mol/L) group. Actin cytoskeleton and ß-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses. RESULTS: In DEX group, there were reorganization of actin cytoskeleton and formation of cross linked actin networks (CLANs), which were partially reversed in DEX and HA1077 group. DEX treatment also induced an increased expression of ß-catenin, which was obviously reduced in DEX and HA1077 group. Meanwhile, the cultured HTM cells in HA1077 group had lower expression of ß-catenin than that in the control group. CONCLUSION: Our results show that HA1077 can reverse the changes of actin organization and expression of ß-catenin induced by DEX in cultured HTM cells, suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.

Novel Compound Heterozygous CBS Mutations Cause Homocystinuria in a Han Chinese Family.

The cystathionine ß-synthase (CBS) gene has been shown to be related to homocystinuria. This study was aimed to detect the mutations in CBS in a Han Chinese family with homocystinuria. A four-generation family from Shandong Province of China was recruited in this study. All available members of the family underwent comprehensive medical examinations. Genomic DNA was collected from peripheral blood of all the participants. The coding sequence of CBS was amplified by polymerase chain reaction (PCR), followed by direct DNA sequencing. Among all the family members, three affected individuals showed typical clinical features of homocystinuria. Two novel compound heterozygous mutations in the CBS gene, c.407T > C (p. L136P) and c.473C > T (p.A158V), were identified by sequencing analysis in this family. Both of the two missense mutations were detected in the three patients. Other available normal individuals, including the patients' parents, grand parents, her younger sister and brother in this family either carried one of the two mutations, or none. In addition, the two mutations were not found in 600 ethnically matched normal controls. This study provides a mutation spectrum of CBS resulting in homocystinuriain a Chinese population, which may shed light on the molecular pathogenesis and clinical diagnosis of CBS-associated homocystinuria.

HOXC10 up-regulation contributes to human thyroid cancer and indicates poor survival outcome.

HOX genes have been well described as important players in development and morphogenesis, and more recently, in carcinogenesis. The role and clinical implication of homeodomain-containing gene 10 (HOXC10) in human thyroid cancer is poorly understood. Real-time PCR and bioinformatics analysis were used to detect the expression of HOXC10 in normal and human thyroid cancer samples from Shanghai General Hospital (also known as Shanghai First People's Hospital) and the TCGA dataset. The survival time of patients with human thyroid cancer was also detected. Cell Count Kit-8 (CCK8) analysis was used to detect cell proliferation, and the cell cycle was assessed by flow cytometry. Migration and invasion were detected by transwell analysis. Gene set enrichment analysis (GSEA) was used to explore the correlation of HOXC10 with signaling pathways. Real-time PCR and Western blot analysis were used to detect the expression of signaling pathway related genes in human thyroid cancer cells. HOXC10 expression in Shanghai General Hospital and the TCGA dataset revealed a significant increase in human thyroid cancer tissues and its expression was positively correlated with the advanced age, poor pathologic stage, and poor prognosis. HOXC10 knockdown by shRNA conferred cell cycle blocking and inhibition of migration and invasion. GSEA in the TCGA datasets revealed that HOXC10 expression was positively correlated with cytokine-cytokine receptor interaction and chemokine signaling pathways. Our data suggest that inhibition of HOXC10 may be a therapeutic strategy for human thyroid cancer treatment. This study investigates the role and clinical implication of HOXC10 in human thyroid cancer.

Isolation and Expression Analysis of CYP9A11 and Cytochrome P450 Reductase Gene in the Beet Armyworm (Lepidoptera: Noctuidae).

Cytochrome P450 monooxygenases (CYPs), as an enzyme superfamily, is widely distributed in organisms and plays a vital function in the metabolism of exogenous and endogenous compounds by interacting with its obligatory redox partner, CYP reductase (CPR). A novel CYP gene (CYP9A11) and CPR gene from the agricultural pest insect Spodoptera exigua were cloned and characterized. The complete cDNA sequences of SeCYP9A11 and SeCPR are 1,931 and 3,919 bp in length, respectively, and contain open reading frames of 1,593 and 2,070 nucleotides, respectively. Analysis of the putative protein sequences indicated that SeCYP9A11 contains a heme-binding domain and the unique characteristic sequence (SRFALCE) of the CYP9 family, in addition to a signal peptide and transmembrane segment at the N-terminal. Alignment analysis revealed that SeCYP9A11 shares the highest sequence similarity with CYP9A13 from Mamestra brassicae, which is 66.54%. The putative protein sequence of SeCPR has all of the classical CPR features, such as an N-terminal membrane anchor; three conserved domain flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN), and nicotinamide adenine dinucleotide phosphate (NADPH) domain; and characteristic binding motifs. Phylogenetic analysis revealed that SeCPR shares the highest identity with HaCPR, which is 95.21%. The SeCYP9A11 and SeCPR genes were detected in the midgut, fat body, and cuticle tissues, and throughout all of the developmental stages of S. exigua. The mRNA levels of SeCYP9A11 and SeCPR decreased remarkably after exposure to plant secondary metabolites quercetin and tannin. The results regarding SeCYP9A11 and SeCPR genes in the current study provide foundation for the further study of S. exigua P450 system.

Synovial Fluid HMGB-1 Levels are Associated with Osteoarthritis Severity.

BACKGROUND: Recent data have shown that enhanced cytokine production in knee osteoarthritis (OA) synovium are believed to promote pathological OA. High-mobility group box 1 (HMGB-1) as a well-known pro-inflammatory cytokine may influence the development of knee OA. The purpose of this study is to analyze the amount of and location of HMGB-1 in OA synovium and to compare it with controls who are afflicted with acute meniscal or cruciate ligament tears. We also evaluated the relationship between the level of HMGB-1 in synovial fluid with the severity of synovitis, clinical symptoms (pain, stiffness, daily activity), and radiological changes in patients with knee OA. METHODS: Synovium and synovial fluid were harvested from seventy-four knee OA patients and thirty-four controls afflicted with acute meniscal or cruciate ligament tears. Kellgren-Lawrence (KL) grading system and Western Ontario McMaster University Osteoarthritis Index (WOMAC) assessment scale were applied to evaluate the radiological and clinic severity of OA patients. Additionally, for all patients, the microscopic synovitis was graded to evaluate the severity of synovium pathology. The location of HMGB-1 was determined in the synovium by immunohistochemistry. Synovium and synovial fluid HMGB-1 levels were measured by western blotting and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: Synovium immunohistochemical analysis revealed that HMGB-1 displayed a strictly nuclear localization in controls; however, both nuclear and cytoplasmic distributions were present in OA patients. The percentage of HMGB-1 positive cells as well as cytoplasmic HMGB-1 cell population in OA patients were higher than those of controls (42.5% vs. 39.7% and 24.0% vs. 5.7%). Both synovium and synovial fluid HMGB-1 levels in OA patients were significantly higher than controls. In OA patients, HMGB-1 in the KL2/3 group was higher than in the KL4 group. Additionally, synovial fluid HMGB-1 levels in OA patients were positively associated with the severity of synovitis, pain, and daily activities. CONCLUSIONS: Our results demonstrated that HMGB-1 is overexpressed and relocated in synovial membranes of patients with knee OA. The increased synovial fluid HMGB-1 levels were associated with the severity of synovitis, pain, and daily activities in knee OA patients. These results suggested that HMGB-1, as a pro-inflammatory cytokine, may play a crucial role in the progression of knee OA.

Cloning and characterisation of NADPH-dependent cytochrome P450 reductase gene in the cotton bollworm, Helicoverpa armigera.

BACKGROUND: Previous studies in our laboratory showed that cytochrome P450 CYP6B7 plays a critical role in a Handan fenvalerate resistant strain (HDFR) of Helicoverpa armigera. As an important component of P450 enzyme systems, cytochrome P450 reductase (CPR) plays an essential role in transferring electrons from NADPH to the P450-substrate complex. However, little information about CPR in H. armigera (HaCPR) has been reported. RESULTS: A full-length cDNA (3525 bp) of HaCPR was cloned. The open reading frame of the HaCPR gene encoded 687 amino acids and shared 27.87-95.21% identities with other known CPRs. Bioinformatic analysis showed that HaCPR is a transmembrane protein with Mw of 77.4 kDa and contains conserved features. The results of real-time quantitative polymerase chain reaction showed that the expression level of HaCPR mRNA was 1.84-fold higher in midgut of 5th instars of the Handan susceptible strain than that in pupae, and the level in the midgut of HDFR strain was 2.02-fold higher than that of the Handan susceptible strain. The levels of HaCPR mRNA were induced by phenobarbital at concentrations of 2 and 4 mg g(-1) , which enhanced 5.20- and 17.45-fold, respectively, compared to that of the control after 48 h of phenobarbital treatment. CONCLUSIONS: The results indicate that HaCPR is important for the development of H. armigera and may play an essential role in the P450-mediated insecticide resistance of H. armigera to fenvalerate.