Solvent Control Group Research Articles

The Effect of Intraurethrally Applied Anatolian Propolis Extract on Urethral Healing in a Rat Model

Purpose: Urethral stricture may cause irreversible results, as it prevents normal voiding. Although various endoscopic and open surgical options are available, the results are not always satisfactory so the main purpose is to prevent the formation of urethral scar. Our purpose was to examine the effects of intraurethrally administered Anatolian propolis on healing after an experimental urethral injury. Materials and Methods: A total of 40 Wistar male rats were used. Rats were divided into five equal groups: healthy control (Group 1), urethral damage/pathology (Group 2), solvent control (Group 3), 1-week propolis treatment (Group 4) and 3-week propolis treatment (Group 5). Urethral damage was performed with a 29G needle. Intraurethral, 50% ethanol, was administered in the solvent control group and 30% propolis was administered intraurethrally to the rats in Groups 3 and 4. Penile tissues were taken under deep anesthesia and examined under a light microscope. Results: Irregularities and luminal narrowing in the urethral epithelium and connective tissue were found in Group 2, except for one rat. Similarly, hyperemia-bleeding was observed in all rats except for one rat. Irregularities and hyperemia in the urethral epithelium and connective tissue were found in Group 3, except for two rats. Total improvement was observed in one rat, and more than 50% of fibrosis was observed in four rats in Groups 2 and 3. In Group 4, irregularity was observed in the urethral epithelium in four rats, while no inflammation was found in five rats. All of the rats had <50% fibrosis. In Group 5, six rats had complete recovery and <30% fibrosis. Conclusion: Anatolian propolis applied into the urethra seems to accelerate recovery after urethral trauma and reduces the formation of fibrosis.

Effects of sorafenib and regorafenib on the expression of hypoxia-inducible factors in hepatocellular carcinoma-transplanted nude mice

Abstract Objective The objective of this study was to investigate the inhibitory effects of sorafenib and regorafenib on the growth of hepatocellular carcinoma (HCC) using a subcutaneous transplantation tumor model in nude mice and exploring the effects of sorafenib and regorafenib on the expression of hypoxia-inducible factor (HIF)-1α, HIF-2α, and HIF-1β in HCC tissues collected from HCC-transplanted nude mice. Methods HepG2 cells were inoculated intradermally into nude mice. The mice were randomly assigned to either sorafenib treatment (100 mg/kg), regorafenib treatment (20 mg/kg), or solvent control group (dimethylsulfoxide) (n = 8 per group) and received once-daily treatment for 14 days. The tumor volumes were recorded every 3 days after the initiation of treatment. The expression levels of HIF-1α, HIF-1β, HIF-2α, and SART1 in the HCC tissues were examined via quantitative real-time PCR (qRT-PCR) analysis and Western blotting. Results The tumors in the sorafenib and regorafenib treatment groups grew slower and smaller than did the tumors in the solvent control group. qPCR analysis and western blotting demonstrated that the mRNA and protein expressions of HIF-1α and HIF-1β were down-regulated. The expression of HIF-2α and SART1 was up-regulated in the sorafenib treatment group (P < 0.05); meanwhile, the expression of HIF-1α and HIF-1β was up-regulated, and that of HIF-2α and SART1 was down-regulated in the regorafenib treatment group (P < 0.05). Conclusion The expression of hypoxia-associated factor is up-regulated by sorafenib and down-regulated by regorafenib, which may induce the different effects of sorafenib on the expression of HIFs.

Oregano Oil Combined with Macleaya Cordata Oral Solution Improves the Growth Performance and Immune Response of Broilers.

Simple SummaryDeveloping safe and effective antibiotic growth promoters (AGPs) substitutes is particularly important to improve animal health and production performance. As an essential plant oil, the oregano oil's main bioactive substance is carvacrol, which has been proven to have antioxidant, anti-inflammatory, antibacterial, and antiviral properties. The sanguinarine from macleaya cordata is the primary bioactive substance. Sanguinarine has anti-tumor, immune-enhancing, antibacterial, and anti-inflammatory effects. However, it has not been reported whether the compatibility of oregano oil and macleaya cordata extract could produce better results. This study is the first to report the effect of oregano oil combined with macleaya cordata oral solution on the growth of broilers. The oregano oil combined with macleaya cordata oral solution significantly improved the growth performance of broilers. At the same time, serum biochemical indices, serum antioxidant indices, serum immune indices, serum cytokines, and intestinal morphology were significantly improved. In summary, our results demonstrated that the mixed solution of oregano oil and macleaya cordata has substantial potential to be an alternative to AGPs for broilers to reduce costs and improve benefits. This study provides basic data and technical support for further research.The abuse of AGPs in animal husbandry has led to severe problems such as drug resistance and ecological, and environmental destruction, which seriously threaten human health and public health security. In recent years, extracts of oregano oil and macleaya cordata have become a hot spot in the research and application of AGP substitutes for their safety and high efficiency. This study is the first to report the effect of oregano oil combined with macleaya cordata oral solution on broiler growth performance. A total of 960 one-day-old broiler chickens were randomly divided into four treatment groups (240 chickens per group). Each treatment group was divided into six replicate groups (40 birds per replicate group). There were four groups in this study: the solvent control group, the oregano essential oil combined with macleaya cordata extract oral solution group (OS group), the oregano essential oil oral solution group (OEO group), and the macleaya cordata extract oral solution group (MCE group). Two chickens from each replicate group were collected and mixed into a composite sample. Six composite samples were obtained for each treatment group. The results showed that the oregano oil combined with macleaya cordata oral solution significantly improved the growth performance of broiler chickens. At the same time, serum biochemical indices, serum antioxidant indices, serum immune indices, serum cytokines, and intestinal morphology were significantly improved by the OS group. This study shows that oregano oil combined with macleaya cordata oral solution has substantial potential to be an alternative to AGPs for broilers.

Effects of octadecadienoic acid on proliferation and apoptosis of glioma cells and its mechanisms

To study the effects of octadecadienoic acid (ODA) on the proliferation and apoptosis of glioma cells and its mechanisms. Cultured human glioma cells (cell density 2×106 cells/L) were divided into solvent control group (DMSO, 30 μl/L), 5-FU group (10 mg/L) and octadecadienic acid groups (0.3, 0.6 and 1.2 mg/L groups). The toxicity of ODA on glioma cells was detected by trypan blue and thiazolium blue (MTT). The expression levels of P53, PI3K, P21, PKB/Akt and Caspase-9 in glioma cells were determined by enzyme-linked immunosorbent assay (ELISA). ① Cell count under optical microscope showed that the inhibition rate of cell proliferation in ODA low, medium and high dose groups and 5-FU group was significantly higher than that in the solvent control group (P＜0.01), but there was no statistical significance compared with the 5-FU group (P＞0.05). ② MTT assay showed that the inhibition rate of cell proliferation was increased significantly in ODA low, medium and high dose groups and 5-FU groups (P＜0.01), compared with the solvent control group. Compared with 5-FU group, the inhibition rate of cell proliferation was increased significantly only in ODA high dose group (P＜0.01). ③ The number of G0/G1 phase cells in ODA low, medium and high dose groups and 5-FU group were increased significantly (P＜0.05, P＜0.01), the number of G2/M phase cells were decreased significantly (P＜0.01), and the apoptosis rate was increased significantly (P＜0.01),compared with the solvent control group. Compared with the 5-FU group, the number of cells in G2/M phase was decreased significantly (P＜0.01) and the apoptosis rate was increased significantly (P＜0.01) in ODA high dose group. ④ ELISA test results showed that the protein expression levels of P53, PI3K and PKB/Akt in ODA low , medium and high dose groups and 5-FU group were significantly lower than those in solvent control group (all P＜0.01), but the protein expression levels in ODA high dose group were significantly lower than those in 5-FU group (P＜0.01). The protein expression levels of P21 and caspase-9 in ODA low , medium and high dose groups and 5-FU group were significantly higher than those in solvent control group (P＜0.05, P＜0.01), but the protein expression levels in ODA high dose group were significantly higher than those in 5-Fu group (P＜0.01). ODA can significantly inhibit the proliferation and promote apoptosis of glioma cells. The mechanisms are related to up-regulating the levels of P21 and caspase-9 to promote apoptosis, down-regulating the levels of P53, PI3K and PKB/Akt to inhibit the cell division cycle, and reducing the activity of PI3K-Akt signal transduction pathway.

Evaluation of Nootropic activity of Bacopa monnieri and Convolvulus pluricaulis herbs extracts with Panchagavya ghrita

Bacopa monnieri and Convolvulus pluricaulis with the Panchagavya ghrita are expected to have better nootropic activity than the combination of Bacopa monnieri and Convolvulus pluricaulis. These marshy herbs are found throughout India having various medicinal importances like treatment of ulcer, epilepsy, memory improvement. The present study was focused on the exploration and assessment of memory and learning activity from the aqueous and methanol extract of Bacopa monnieri (Brahmi) and Convolvulus pluricaulis (Sankhapuspi) leaves with and without the Panchagavya ghrita (PGG) by performing different pharmacological screening models. The study of nootropic activity was performed by using different screening methods such as; phenobarbitone sodium-induced sleeping time, elevated plus maze, radial arm maze, actophotometer. Both methanol and aqueous extracts of Brahmi, Sankhapuspi with the combinations of Panchagavya ghrita showed a significant increase in memory and learning activity as compared to solvent control group where methanol extract containing combination exhibited better enhancement of memory learning activity in all experimental models as compared to aqueous extract. However, it is confirmed that herbs are having excellent activities in learning and memory when combined with Panchagavya ghrita.

Effect of Omega-3-Rich Fish Oil on TNF-  Expression in Mice's Colonic Tissue Induced with Azoxymethane (AOM) and Dextran Sodium Sulphate (DSS)

Objective: Colorectal cancer (CC) is one type of cancer with a high incidence worldwide. Many therapeutic techniques have been for CC but have not yet yielded satisfactory results. Fish oil has potential as an alternative therapy for CC through its anti-inflammatory effects. Here, we want to investigate that effect on TNF-α expression using omega-3-rich fish oil (FO). Methods: FO was injected into Swiss mice that have been induced (2% in drinking water) with DSS for seven consecutive days. Animals were separated into six groups: normal, negative control, positive control, solvent control, and FO groups (3 and 6 mg/kg body weight/day). All animals were sacrificed, and the colons were collected then stained with anti-TNF-α. The stained sections were subsequently examined with ImageJ based on colour density. Results: Based on the H-Score of each group, FO 3 mg and 6 mg has significantly decreased the expression of TNF-α compared to the negative control (p=0.001 and p=0.009). Conclusion: FO administration was able to inhibit the expression of TNF-α on mice's colonic tissue induced with AOM and DSS.

The role of TNF-α/TNFR1 inhibiting autophagy of M1-type Kupffer cells in liver injury in trichloroethylene-sensitized mice

Objective: To detect the expression levels of M1-type polarization and autophagy-related indicators in the liver of trichloroethylene (TCE) -sensitized mice, and to explore the role of liver tumor necrosis factor-α (TNF-α) and tumor necrosis factor receptor 1 (TNFR1) in regulating M1-type Kupffer cells autophagy in liver injury in TCE-sensitized mice. Methods: In November 2019, according to simple random grouping, 45 SPF grade BALB/c female mice (6-8 weeks old) were divided into 4 groups: blank control group (n=5) , solvent control group (n=5) , TCE treatment group (n=18) , TCE+R7050 (inhibitor) treatment group (n=17) . Transdermally sensitized mice, 24 h after the last challenge, the mice were divided into TCE sensitized group and TCE non-sensitized group according to the skin reaction score. The livers of mice were harvested, and the pathological changes of the livers were observed under light and electron microscopes. Western blotting was used to detect the expressions of TNF-α, TNFR1 and autophagy-related indexes. The expression of inducible nitric oxide synthase (iNOS) , a marker of M1-type Kupffer cells, was detected by immunohistochemistry, and the occurrence of autophagy in M1-type Kupffer cells was detected by immunofluorescence double-labeling method. Results: The sensitization rate of TCE treatment group was 38.9% (7/18) , and TCE+R7050 treatment group was 35.3% (6/17) , with no significant difference between the two groups (P=1.000) . Compared with the blank control group, mice in the TCE sensitized group had abnormal liver ocytes, obvious liver injury, reduced mitochondria and broken endoplasmic reticulum. Western blotting results showed that the expressions of TNF-α and TNFR1 protein in the liver of the mice in the TCE sensitized group increased, the expression of iNOS protein in M1-type Kupffer cells increased, and the expressions of autophagic microtubule-associated protein 1 light-chain 3 (LC3B) and Beclin1 protein were decreased (P<0.05) . The results of immunohistochemistry showed that iNOS was not significantly expressed in the blank control group and solvent control group, and a small amount of expression was found in the TCE non-sensitized group, the positive staining area was obvious in TCE sensitized group, and the expression of iNOS was significantly increased (P<0.05) . Immunofluorescence results showed that the iNOS protein levels in the blank control group, solvent control group and TCE non-sensitized group were lower, and only partially colocalized with P62; the colocalization of iNOS with P62 in the TCE sensitized group was significantly increased. Conclusion: TNF-α/TNFR1 signaling pathway may promote liver injury in TCE-sensitized mice by inhibiting autophagy of M1-type Kupffer cells.

Identification of pathways and key genes in male late-stage carotid atherosclerosis using bioinformatics analysis.

Late-stage carotid atherosclerosis has a high incidence rate and may lead to various cerebrovascular diseases. The gene expression profile GSE100927 was selected to identify differentially expressed genes (DEGs) in carotid atherosclerosis. Subsequently, protein-protein interaction, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were conducted. Furthermore, experimental verification was performed using human umbilical vein endothelial cells (HUVECs), human aortic vascular smooth muscle cells (HAVSMCs) and Tohoku Hospital Pediatrics-1 (THP-1)-induced macrophages. The groups were as follows: Control group, solvent control group and palmitic acid group. The levels of reactive oxygen species (ROS) in the three cell types were detected by flow cytometry or fluorescence microscopy. Furthermore, apoptosis of HUVECs and HAVSMCs was assessed by flow cytometry and the nuclear Hoechst 33258 staining of THP-1-induced macrophages was performed. Male late-stage carotid atherosclerosis samples, including 10 control samples and 21 atherosclerosis samples, were selected. Pathway enrichment analysis demonstrated that ‘Toll-like receptor signaling pathway’ was the top pathway associated with the DEGs. MMP7, MMP9, IL1β, C-C motif chemokine ligand 4 (CCL4), secreted phosphoprotein 1 (SPP1), CCL3 and interferon regulatory factor 5 (IRF5) were selected for experimental verification. Palmitic acid increased the ROS levels and the apoptosis rates of HUVECs and HAVSMCs. However, it did not increase the levels of ROS and did not shrink the nuclei of THP-1-induced macrophages. Furthermore, palmitic acid increased the mRNA levels of IL1β, CCL4, SPP1, CCL3, IRF5, MMP7 and MMP9 in HUVECs and THP-1-induced macrophages, and increased the mRNA levels of CCL4 and MMP9 in HAVSMCs. In conclusion, IL1β, CCL3, CCL4, SPP1, IRF5, MMP7 and MMP9 are important markers of late-stage carotid atherosclerosis.

The role of ROS/TXNIP/NLRP3 pathway in the skin injury of trichloroethylene sensitized mice

Objective: To explore the mechanism of reactive oxygen species/thioredoxin-interacting protein/nucleotide-binding oligomerization domain-like receptor 3 (ROS/TXNIP/NLRP3) pathway in the skin injury of trichloroethylene (TCE) sensitized mice. Methods: In August 2020, 40 female BALB/c mice were randomly divided into control group (n=5) , solvent control group (n=5) , TCE treatment group (n=15) and TCE+(2-(2, 2, 6, 6-Tetrameyhylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride (Mito TEMPO) treatment group (n=15) . The TCE sensitization model was established. Mice in the TCE treatment group and TCE+Mito TEMPO treatment group were divided into the sensitized positive group and the sensitized negative group according to the skin erythema and edema reactions on the back of the mice 24 h after the last stimulation. The mice were sacrificed 72 h after the last stimulation, the back skin of the mice was taken, and the skin lesions were observed. Immunohistochemistry (IHC) was used to detect the expression level of NLRP3, and the Western Blot was performed to detect the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) , cysteinyl aspartate specific proteinase 1 (Caspase 1) , Interleukin-1β (IL-1β) and TXNIP proteins in the skin of the mice, the reactive oxygen species (ROS) kit was used to detect the level of intracellular ROS in the back skin tissue. Results: The sensitization rates of TCE treatment group and TCE+Mito TEMPO treatment group were 40.0% (6/15) and 33.3% (5/15) , respectively, and there was no significant difference between the two groups (P>0.05) . The back skin of the mice in the TCE sensitized positive group was thickened and infiltrated by a large number of inflammatory cells. The number of mitochondria in the epidermis cells was significantly reduced, the mitochondrial crest disappeared and vacuolar degeneration occurred. TCE+Mito TEMPO sensitized positive group had less damage, more mitochondria and relatively normal cell structure. Compared with the solvent control group and corresponding sensitized negative groups, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE sensitized positive group and TCE+Mito TEMPO sensitized positive group were significantly increased (P<0.05) . Compared with TCE sensitized positive group, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE+Mito TEMPO sensitized positive group were significantly decreased (P<0.05) . Conclusion: ROS/TXNIP/NLRP3 pathway was activated and then encouraged the release of IL-1β, finally aggravated the TCE-induced skin injury.

Combined effect of diazepam and polystyrene microplastics on the social behavior of medaka (Oryzias latipes)

The combined effect of microplastics and pharmaceuticals on aquatic organisms is an issue of concern. In this laboratory study, we evaluated the combined effect of polystyrene microplastics (2-μm diameter) and diazepam on the social behavior of medaka (Oryzias latipes) by using the shoaling behavior test with five treatment groups: solvent control, polystyrene microplastics exposure (0.04 mg/L), low-concentration diazepam exposure (0.03 mg/L), high-concentration diazepam exposure (0.3 mg/L), and polystyrene microplastics and low-concentration diazepam co-exposure. After 7 days of exposure, the shoal-leaving behavior of the high-concentration diazepam exposure group (8.9 ± 8.3 counts/medaka) and the co-exposure group (6.8 ± 6.7 counts/medaka) was significantly greater than that in the solvent control group (1.8 ± 2.6 counts/medaka). Even after 5 days of recovery, medaka in the co-exposure group left the shoal more often (7.3 ± 5.0 counts/medaka) than those in the solvent control group (2.6 ± 2.6 counts/medaka), whereas the shoal-leaving behavior in other exposure groups, except for the high-concentration diazepam exposure group, was restored. Our findings show that the combined effects of diazepam and polystyrene microplastics suppressed medaka social behavior, suggesting that the presence of microplastics can enhance the adverse effects of pollutants on the social behavior of aquatic organisms.

General toxicity and genotoxicity of altertoxin I: A novel 28-day multiendpoint assessment in male Sprague-Dawley rats.

The mycotoxin altertoxin I (ATX-I) is one of secondary metabolites produced by Alternaria fungi and is frequently detected as food and feed contaminants. Little is known about the genotoxicity of the ATX-I. In order to evaluate potential genotoxicity and general toxicity of ATX-I, the novel 28-day multiendpoint (Pig-a assay + micronucleus [MN] test + comet assay) genotoxicity platform was applied. Male Sprague-Dawley (SD) rats were randomized to five groups (six rats per group), that is, a positive control group (N-ethyl-N-nitrosourea [ENU], 40 mg/kg.bw/d), two solvent control groups (PBS and corn oil), and two ATX-I-treated groups (low-dose group [1.10μg/kg.bw/d] and high-dose group [5.51 μg/kg.bw/d]). Treatments were administered by oral gavage to male SD rats for 28 consecutive days. Histopathological damages in the liver, kidney, and spleen were observed, but without significant changes in hematological and serum biochemical parameters. Genotoxic endpoints indicated that ATX-I could cause DNA damage. To summarize, in a relatively low-dose range, ATX-I may not have direct genotoxicity in vivo but could induce liver, kidney, and spleen damage.

The evaluation of cytotoxicity and cytokine IL-6 production of root canal sealers with and without the incorporation of simvastatin: an invitro study

BackgroundFreshly mixed root canal sealers when proximate the periapical tissues, trigger varying degrees of cytotoxicity/inflammatory reactions. Simvastatin, a class of the drug statin, is a widely used cholesterol-lowering agent with additional anti-inflammatory activities. This study assessed the effects of simvastatin on cytotoxicity and the release of IL-6 (Interleukin-6) production when incorporated in zinc oxide eugenol and methacrylate resin-based sealers.MethodsExperimental groups consisted of conventional zinc oxide eugenol and methacrylate based-EndoREZ sealers (ZE & ER respectively) and 0.5 mg/mL simvastatin incorporated sealers (ZES & ERS). L929 mouse fibroblast cells were exposed to freshly mixed experimental sealers and evaluated for cytotoxicity (MTT assay) and inflammation levels (inflammatory marker IL-6 for ELISA) at various time intervals (0h, 24h and 7th day). The values were compared to the cell control (CC; L929 cells alone) and solvent control (SC; L929 cells + DMSO) groups. All the experiments were conducted in triplicates and subjected to statistical analysis using IBM SPSS Statistics software. Non parametric tests were conducted using Kruskal-Wallis and Friedman tests for inter-group and intra-group comparisons respectively. Pairwise comparison was conducted by post hoc Dunn test followed by Bonferroni correction. P values < 0.05 were considered statistically significant.ResultsAll the experimental groups (ZE, ER, ZES, ERS) exhibited varying degree of cytotoxicity and IL-6 expression compared to the control groups CC and SC. The cell viability for ZE and ER decreased on day 7 as compared to 24 h. ZES and ERS had higher viable cells (75.93% & 79.90%) compared to ZE and ER (54.39% & 57.84%) at all time periods. Increased expression of IL-6 was observed in ZE & ER (25.49 pg/mL & 23.14 pg/mL) when compared to simvastatin incorporated ZE & ER (ZES-12.70 pg/mL & ERS-14.68 pg/mL) at all time periods. Highest level of cytotoxicity and inflammation was observed in ZE compared to all the other groups on day 7.ConclusionsAddition of 0.5 mg/mL of simvastatin to the sealers (ZES and ERS) decreased the cytotoxicity in the freshly mixed state and reduces their inflammatory effect.

EVALUATION OF PTEN AND PI3K/AKT EXPRESSIONS IN STANOZOLOL-TREATED RAT KIDNEYS

Objective: Stanozolol is an anabolic androgenic steroid (AAS) that is widely used by teenagers and athletes in bodybuilding, sports, and athletics. The potential effects of stanozolol in kidney functions have not been defined. In this study we investigated the expression of tumor suppressor protein phosphatase and tensin homolog (Pten) and mRNA levels of phosphatidylinositol 4,5-bisphosphate 3 kinase (Pi3k) and the protein kinase B (Akt1) signaling pathway in rat kidneys treated by stanozolol. Material and Method: Rats were randomized to 5 groups as control, solvent control, steroid (stanozolol), solvent-exercise, and steroid-exercise. Subcutaneous injection of stanozolol (5 mg/kg) was applied for 28 days and swimming exercise was performed 20 min/day, 5 days/week in exercise groups. Expression of PTEN was evaluated by immunohistochemistry. Also, Pten, Pi3k, and Akt1 mRNA expressions were analyzed via RT-PCR. Results: Mesangial cells and renal tubules in the control, solvent control, and solvent exercise groups showed strong (+++) PTEN reactivity against weak PTEN reactivity in the steroid group. Moderate PTEN reactivity was detected in cells of the steroid exercise group. Pten mRNA expression was significantly decreased, and Pi3k and Akt1 mRNA expression were significantly increased in the steroid group versus other groups (p&lt;0.001). Pten expression showed increase while Pi3k and Akt1 expression showed decrease with exercise treatment (p&lt;0.05). Conclusion: Our findings suggest that AAS usage may inhibit PTEN expression in kidneys, which can be associated with increased Pi3k and Akt1 mRNA levels. Exercise performed with AAS usage can be protective on stanozolol-exposed kidneys due to increased levels of PTEN expression and decreased levels of Pi3k and Akt1.

EVALUATION OF PTEN AND PI3K/AKT EXPRESSIONS IN STANOZOLOL-TREATED RAT KIDNEYS

Objective: Stanozolol is an anabolic androgenic steroid (AAS) that is widely used by teenagers and athletes in bodybuilding, sports, and athletics. The potential effects of stanozolol in kidney functions have not been defined. In this study we investigated the expression of tumor suppressor protein phosphatase and tensin homolog (Pten) and mRNA levels of phosphatidylinositol 4,5-bisphosphate 3 kinase (Pi3k) and the protein kinase B (Akt1) signaling pathway in rat kidneys treated by stanozolol. Material and Method: Rats were randomized to 5 groups as control, solvent control, steroid (stanozolol), solvent-exercise, and steroid-exercise. Subcutaneous injection of stanozolol (5 mg/kg) was applied for 28 days and swimming exercise was performed 20 min/day, 5 days/week in exercise groups. Expression of PTEN was evaluated by immunohistochemistry. Also, Pten, Pi3k, and Akt1 mRNA expressions were analyzed via RT-PCR. Results: Mesangial cells and renal tubules in the control, solvent control, and solvent exercise groups showed strong (+++) PTEN reactivity against weak PTEN reactivity in the steroid group. Moderate PTEN reactivity was detected in cells of the steroid exercise group. Pten mRNA expression was significantly decreased, and Pi3k and Akt1 mRNA expression were significantly increased in the steroid group versus other groups (p&lt;0.001). Pten expression showed increase while Pi3k and Akt1 expression showed decrease with exercise treatment (p&lt;0.05). Conclusion: Our findings suggest that AAS usage may inhibit PTEN expression in kidneys, which can be associated with increased Pi3k and Akt1 mRNA levels. Exercise performed with AAS usage can be protective on stanozolol-exposed kidneys due to increased levels of PTEN expression and decreased levels of Pi3k and Akt1.

The Role and Mechanism of M2c Macrophages in Lipopolysaccharide/D-Galactosamine Induced Acute Hepatitis

The study was based on the evaluation of the positive effect of M2c macrophages on liver tissue inflammation, pathological changes and tissue damage in lipopolysaccharide/D-galactosamine induced acute hepatitis. Meanwhile, whether M2c macrophages induced anti-inflammatory effects through Janus kinase/signal transducer and activator of transcription signaling pathway was also investigated. Forty institute of cancer research female mice in the same batch with no significant difference in body weight were distributed randomly into 5 different groups namely the control group, solvent control group, lipopolysaccharide/D-galactosamine, lipopolysaccharide/D-galactosamine+M2c and lipopolysaccharide/ D-galactosamine+M2c+ruxolitinib, 8 in each group. 6 h after the last treatment of mice in each group, serum was collected to detect the content of alanine aminotransferase and aspartate transferase. The pathology of liver tissue was determined using hematoxylineosin staining. The determination of the levels of expression of anti-inflammatory factors, pro-inflammatory factors, related apoptosis proteins and Janus kinase/signal transducer and activator of transcription signaling pathway related proteins in liver tissue were carried out by Western blot. After 6 h of induction with lipopolysaccharide/D-galactosamine, the liver function (alanine aminotransferase, aspartate aminotransferase), pathological score of liver slices, liver pro-inflammatory factors and anti-inflammatory factors (interleukin-6, tumor necrosis factor-alpha, interleukin-1beta and interleukin-10) were significantly improved in the mice blood. Comparative to the normal control group, apoptosis-related proteins (B-cell lymphoma-extra-large, B-cell lymphoma 2, Bcl-2- associated X protein) and other indicators were higher with a significant difference (p<0.05) under the acute hepatitis model induced lipopolysaccharide/D-galactosamine, after infusion of M2c macrophages from the tail vein of mice. The above indexes of mice were improved and exhibited a statistically significant difference (p<0.05). After adding ruxolitinib to intervene Janus kinase/signal transducer and activator of transcription signaling pathway on the basis of lipopolysaccharide/D-galactosamine+M2c group, the liver function, liver slice pathological score, pro and anti-inflammatory factor and apoptosis-related protein expressions were significantly higher as compared to the lipopolysaccharide/D-galactosamine+M2c group with significant difference (p<0.05), while the observed differences were not significant (p>0.05) in the lipopolysaccharide/ D-galactosamine group. Lipopolysaccharide/D-galactosamine induces the occurrence of acute hepatitis, which can well mimic the virus the pathogenic process of hepatitis. M2c macrophages exhibited a critical anti-inflammatory role in acute hepatitis via activating the Janus kinase/signal transducer and activator of transcription signaling pathway.

Assessing sub-lethal effects of the dinitroaniline herbicide pendimethalin in zebrafish embryos/larvae (Danio rerio)

Pendimethalin is a dinitroaniline herbicide used to control broadleaf weeds by inhibiting the formation of microtubules during cell division. Its use on a variety of crops leads to its potential entry into aquatic environments, but little is known about its sub-lethal toxicity to early developmental stages of aquatic vertebrates. To address this knowledge gap, we assessed the toxicity of pendimethalin to zebrafish embryos and larvae by measuring mortality, developmental abnormalities, oxidative respiration, reactive oxygen species, gene expression, and locomotor activity following exposure to the herbicide throughout early development. Embryos at ~6 h post-fertilization (hpf) were exposed to either a solvent control (0.1% DMSO, v/v), embryo rearing medium (ERM), or one dose of either 1, 2.5, 5, or 25 μM pendimethalin for up to 7-days post fertilization depending on the bioassay. Exposure to 25 μM pendimethalin resulted in high prevalence of spinal curvature, tail malformations, pericardial edema, and yolk sac edema at 4 dpf, while exposure to 5 μM pendimethalin induced pericardial edema and lordosis in the fish exposed over 7 dpf. Exposure to pendimethalin up to 5 μM did not negatively impact oxidative respiration (e.g., basal respiration, oligomycin-induced ATP production) in embryos following a 24-h exposure. Pendimethalin did not induce reactive oxygen species at concentrations of 1–5 μM. Levels of transcripts associated with oxidative respiration and damage response were altered in 7d-larvae; cox1 mRNA was increased in larvae fish exposed to 1 μM while cox5a1 and sod2 mRNA were decreased with 2.5 μM exposure. The Visual Motor Response (VMR) test for light-dark response revealed that larval activity in the dark period was reduced for zebrafish exposed to >1 μM pendimethalin compared to ERM and DMSO solvent control groups. These data inform on the sub-lethal toxicity of pendimethalin to early stages of fish embryos and larvae.

Nigella Sativa’s Protection Against 7,12 Dimethylbenz [A] Anthracene -Induced Colon Carcinogenesis in Rats

Colon cancer is the third most common cause of death from cancer worldwide. Recently, natural products have been widely used as an alternative therapy for colon cancer. Previous studies have reported that Nigella sativa has chemopreventive activity in vitro and in vivo.This study aimed to evaluate the effect of Nigella sativa seed (NSS) on rat-colon cell after initiation of 7,12-dimethylbenz [a] anthracene. Rats were divided into five groups, 12 rats in each group: Group I was given 7,12dimetilbenz [a] anthracene (DMBA) orally 20 mg/kgBW twice a week for five weeks, group V is the solvent control group was given corn oil. The other three groups were given DMBA + NSS, at the dosage of 250 mg/kgBW, 500 mg/kgBW and 750 mg/kgBW. NSS extract was dissolved in corn oil and administered daily per oral during the next two weeks before and during the initiation of DMBA. After 16 weeks, all rats were sacrificed. H&amp;E staining showed that necrosis activity was lower in treated groups compared to DMBA group. AgNOR staining showed mAgNOR was significantly decrease following the increasing dose of NSS (250 mg/kgBW, 500 mg/kgBW and 750 mg/kgBW) were subsequently 1.62 ± 0.086, 1.60 ± 0.101 and 1.39 ± 0.049 (p&lt;0.05). The results showed that NNS reduce the damage of colon cells and inhibit colon cell proliferation in DMBA induced rats.

Role of LncRNA CASC11 in the malignant transformation of 16HBE cells induced by glycidyl methacrylate

To investigate the effect of trending up-regulation LncRNA CASC11 which is differentially expressed during the malignant transformation of human bronchial epithelial cells(16 HBE) induced by glycidyl methacrylate(GMA). After 16 HBE cells were repeatedly exposed to low concentration GMA(8 μg/mL), the 10 th, 20 th and 30 th passage cells of the GMA group and the DMSO solvent control group were collected. High throughput LncRNA microarray were used to screen the differentially expressed LncRNAs at different stages. Short Time-series Expression Miner and bioinformatics tool RPISeq were used to screen and predict the potential targets of specific LncRNA. Real-time fluorescent quantitative PCR(qPCR) and Western blot were used to detect the relative expression of specific LncRNA and the content of its target protein respectively. The specific differential expression of LncRNA CASC11 in the process of GMA-induced malignant transformation of 16 HBE cells showed a trend of up-regulation. Compared with the control cells in the same period, the P10, P20 and P30 generation cells were down-regulated by 1.64 times, up-regulated by 2.01 times, and up-regulated by 2.66 times, respectively. Western blot showed that the levels of cyclin-dependent kinase 1(CDK1), cyclinB1 and cyclinB2 in P10, P20 and P30 cells after exposure were lower than those in DMSO control group during the same period, which was consistent with the microarray results. The differential expression of LncRNA CASC11 in the process of GMA-induced 16 HBE cell malignant transformation was up-regulated trendingly. It is speculated that it may block or delay cell cycle progression by inhibiting CDK1.

Phytoestrogens on the Proliferation of Breast Cancer Cell MCF-7

A cell is an organic whole containing biological macromolecules such as nucleic acid and protein. Human cells contain about 15,000 proteins, which play an important role in the development of cell life. This article aims to study the effect of phytoestrogens on the proliferation of breast cancer cells MCF-7. The estrogen-dependent MCF-7 cells described in this article are grown on average DMEM (containing calf serum 10) and connected to single cell culture. Five days before adding the test product, the cells were washed with PBS and converted into red-free phenol. High-sugar DMEM containing 5 parts of bovine embryo serum was cultured and treated with activated charcoal glucan glycosides. In the experiment, four dose groups of solvent control, estrogen control, anti-estrogens and two test substances were used for control, and the proliferation of MCF-7 cells was analyzed by integration method and flow cytometry. The experimental results in this article show that compared with the solvent control group, GS can significantly inhibit MCF-7 cell proliferation and cell DNA synthesis, and G/M can block the cell cycle and produce a similar inhibitory effect. 96mol/L zein treatment for 24 hours can significantly promote MCF-7 cell proliferation and cell DNA composition, promote cell cycle, and increase cell separation and proliferation index.

Study on the effect and mechanism of recombinant human intestinal trefoil factor on intestinal mucosal injury and repair in burned mice

Study on the effect and mechanism of recombinant human intestinal trefoil factor on intestinal mucosal injury and repair in burned mice