The effect of methyltestosterone (MT) on sex differentiation and growth in juvenile yellow perch (Perca flavescens).

A study was conducted to evaluate the gonad differentiation of juvenile yellow perch (YP, Perca flavencens) and determine the latest labile period related to hormone treatment. Juvenile fish were subjected to two dietary concentrations of methyltestosterone (MT; 20 and 50 mg/kg feed) for 60 days in three (3) age groups of 38-, 46-, and 67-days post-hatching (dph), where control group were fed with standard commercial feed. Following a 10-month on-growing period, sex phenotypes were determined by gross and histological gonad morphology. Results showed the juvenile YP responded to the exogenous hormone when it was applied at 38 dph for both 20 and 50 mg/kg feed resulting in 100% males. At 46 dph, only 50 mg/kg feed resulted in 100% males. Both MT-treated at 38 and 46 dph significantly differed (P < 0.01) from the expected normal population of male:female (1:1). MT-treated at 67 dph resulted in 37% and 25% intersex fish for both 20 and 50 mg/kg feed dosage groups, respectively. MT-treated at 38 and 46 dph promoted growth and showed significantly heavier mean body weight (P < 0.05) compared to control. The gonadosomatic index (GSI) of MT-treated at 38 and 46 dph was significantly lower than that in control. This study provides the first evidence that juvenile YP can be successfully masculinized when the treatment is initiated at the age of up to 46 dph. The result is important for sex control in aquaculture.

Ginsenoside regulates Treg/Th17 cell ratio and inhibits inflammation to treat COPD.

Objective: Several studies have suggested an involvement of the immune system in the occurrence and development of chronic obstructive pulmonary disease (COPD), but the mechanism is still unclear. The aim of this study was to explore the mechanism of ginsenoside in inhibiting inflammation by regulating FOXP3 in COPD. Methods: Eighty COPD patients were selected and 35 healthy people were enrolled in the study to determine clinical efficacy, observation index, and SGRQ scores. Percentage of Treg and Th17 cells were detected by flow cytometry; HE staining was used to detect the effect of ginsenoside therapy on pathological changes of COPD in mice. Additionally, we transfected FOXP3 inhibitor; RT-PCR and western blot were used to detect the inflammation related genes and proteins. Results: The basic information of the patients were comparable. The clinical outcome in the treatment group was better than that in the control group, which indicated that ginsenoside has a certain therapeutic effect on COPD patients. The lung function and 6MWT distance results indicated that ginsenoside could stabilize the clinical symptoms of COPD patients and improve their quality of life. Flow cytometry results showed that ginsenoside can increase Treg expression while reducing Th17 cell expression. RT-PCR and western blot results showed that the expression of TNF-α and IL-17 in the model group was significantly increased after treatment, obviously caused by an increased expression of FOXP3. Conclusion: Ginsenoside can inhibit inflammation in COPD by up-regulating FOXP3.

Onset of Coronary Heart Disease is Associated with HCMV Infection and Increased CD14 +CD16 + Monocytes in a Population of Weifang, China.

OBJECTIVE: To investigate the relationship between human cytomegalovirus (HCMV) infection and peripheral blood CD14 +CD16 + monocytes in the pathogenesis of coronary heart disease (CHD), and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection, inflammation, and CHD, to provide a basis for the prevention, evaluation, and treatment of the disease. METHODS: In total, 192 patients with CHD were divided into three groups: latent CHD, angina pectoris, and myocardial infarction. HCMV-IgM and -IgG antibodies were assessed using ELISA; CD14 +CD16 + monocytes were counted using a five-type automated hematology analyzer; mononuclear cells were assessed using fluorescence-activated cell sorting; and an automatic biochemical analyzer was used to measure the levels of triglyceride, cholesterol, high- and low-density lipoprotein cholesterols, lipoprotein, hs-CRp and Hcy. RESULTS: The positive rates of HCMV-IgM and -IgG were significantly higher in the CHD groups than in the control group. HCMV infection affects lipid metabolism to promote immune and inflammatory responses. CONCLUSION: HCMV infection has a specific correlation with the occurrence and development of CHD. The expression of CD14 +CD16 + mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection. Thus, HCMV antibody as well as peripheral blood CD14 +CD16 + mononuclear cells can be used to monitor the occurrence and development of CHD.

Effects of intraperitoneal hydrogen injection on nitric oxide synthase mRNA and malondialdehyde following limb ischemia-reperfusion in rabbits.

OBJECTIVE: The purpose of this study was to investigate the effects of intraperitoneal hydrogen (H2) injection on the mRNA expression levels of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) as well as the serum malondialdehyde (MDA) level in a rabbit model of limb ischemia-reperfusion (I/R)-induced skeletal muscle injury. METHODS: To establish the hind limb I/R animal model, 30 rabbits were randomly assigned to one of 3 groups: sham, I/R, and ischemia-reperfusion + H2 (IRH). An intraperitoneal injection of H2 was given to the IRH group, while an equivalent amount of air was given to the sham and I/R groups. At 3, 6, 12, and 24 h after reperfusion, serum MDA level as well as skeletal muscle iNOS and eNOS mRNA expression levels were determined. RESULTS: Both iNOS mRNA expression and serum MDA levels were higher in the I/R group than the sham group (p<0.01) and lower in the IRH group than the I/R group (p<0.01, p<0.05, respectively) at various time points after reperfusion. The eNOS mRNA expression level exhibited no significant difference between the I/R and sham groups after reperfusion but was significantly higher in the IRH group than in the sham group (p<0.01, p<0.05, respectively). CONCLUSION: During the I/R process, the expression of iNOS mRNA was up-regulated along with an increase in MDA. Intraperitoneal injection of H2 can down-regulate iNOS mRNA expression and up-regulate eNOS mRNA expression in the I/R process, suggesting a protective effect of H2 in I/R-induced skeletal muscle injury.

Down-regulation of aquaporin3 expression by lipopolysaccharide via p38/c-Jun N-terminal kinase signalling pathway in HT-29 human colon epithelial cells.

AIM: To investigate the influence of lipopolysaccharide (LPS) through the p38/c-Jun N-terminal kinase (JNK) signalling pathway on aquaporin 3 (AQP3) expression in HT-29 human colon epithelial cells. METHODS: HT-29 cells were treated with LPS, and then the membrane localisation of AQP3 was examined by immunofluorescence staining. The mRNA and protein expression of AQP3 with LPS exposure was measured by real-time reverse transcription-PCR and Western blot, respectively. Activation of p38 and JNK was evaluated by detection of phosphorylation of p38 and JNK using Western blot assay. AQP3 protein expression was determined by Western blot in cells after treatment with SB203580, a selective p38 MAPK inhibitor, or SP600125, a selective JNK inhibitor. RESULTS: In HT-29 cells, the transcription and protein expression of AQP3 were decreased by LPS in a dose- and time-dependent manner, the expression of AQP3 was significantly decreased with the increased concentration of LPS, and at a dose of 100 µg/mL LPS, AQP3 mRNA and protein levels were decreased by a maximum (P < 0.05) of 1.51-fold and 1.49-fold, respectively. When cells were treated with 100 µg/mL LPS for 0, 3, 6, 12, and 24 h, the AQP3 mRNA level was significantly decreased at an early time point of 3 h, and reached about 10% of the control level at 24 h post-treatment (P < 0.05). Down-regulation of AQP3 expression was significantly inhibited by the p38 inhibitor (SB203580) and JNK inhibitor (SP600125). CONCLUSION: p38 and JNK may be promising targets for the preservation of AQP3 expression and may be beneficial to the clinical management of diarrhoea.

Molecular cloning and mRNA expression of the peptidoglycan recognition protein gene HcPGRP1 and its isoform HcPGRP1a from the freshwater mussel Hyriopsis cumingi.

Peptidoglycan recognition proteins (PGRPs) are innate immune molecules that have been structurally conserved throughout evolution in invertebrates and vertebrates. In this study, peptidoglycan recognition protein HcPGRP1 and its isoform HcPGRP1a were identified in the freshwater mussel Hyriopsis cumingii. The full-length cDNAs of HcPGRP1 (973 bp) and HcPGRP1a (537 bp) encoded polypeptides with 218 and 151 amino acids, respectively. Sequence analysis showed that HcPGRP1 had one C-terminal PGRP domain that was conserved throughout evolution. Phylogenetic analysis showed that HcPGRP1 clustered closely with EsPGRP4 of Euprymna scolopes. Real-time PCR showed that the mRNA transcripts of HcPGRP1 and HcPGRP1a were constitutively expressed in various tissues, with the highest level in hepatopancreas. Stimulation with lipopolysaccharide (LPS) and peptidoglycan (PGN) significantly up-regulated HcPGRP1 mRNA expression in hepatopancreas and foot, but not in gill, whereas HcPGRP1a expression was significantly up-regulated in all three tissues. Our results indicate that HcPGRP1 is both a constitutive and inducible protein that may be involved in immune responses (recognition and defense) against invaders.

Quantitative guidelines for the prediction of ultrasound contrast agent destruction during injection.

Experiments and theory were undertaken on the destruction of ultrasound contrast agent microbubbles on needle injection, with the aim of predicting agent loss during in vivo studies. Agents were expelled through a variety of syringe and needle combinations, subjecting the microbubbles to a range of pressure drops. Imaging of the bubbles identified cases where bubbles were destroyed and the extent of destruction. Fluid-dynamic calculations determined the pressure drop for each syringe and needle combination. It was found that agent destruction occurred at a critical pressure drop that depended only on the type of microbubble. Protein-shelled microbubbles (sonicated bovine serum albumin) were virtually all destroyed above their critical pressure drop of 109 ± 7 kPa Two types of lipid-shelled microbubbles were found to have a pressure drop threshold above which more than 50% of the microbubbles were destroyed. The commercial lipid-shelled agent Definity was found to have a critical pressure drop for destruction of 230 ± 10 kPa; for a previously published lipid-shelled agent, this value was 150 ± 40 kPa. It is recommended that attention to the predictions of a simple formula could preclude unnecessary destruction of microbubble contrast agent during in vivo injections. This approach may also preclude undesirable release of drug or gene payloads in targeted microbubble therapies. Example values of appropriate injection rates for various agents and conditions are given.

Melatonin ameliorates bisphenol A-induced DNA damage in the germ cells of adult male rats.

Bisphenol A (BPA) is a well-known endocrine-disrupting chemical (EDC) that has received particular attention because of its widespread distribution in humans. Due to its chemical similarity to diethylstilbestrol, which is carcinogenic to mammals, the possible genotoxicity of BPA has already largely been evaluated. However, the results are still inconclusive and controversial. To investigate the genotoxic effects of BPA in rat germ cells and the potential protective action of melatonin against these effects, adult male Sprague-Dawley rats were orally administered BPA at a dose of 200mg/kg body weight per day for ten consecutive days with or without melatonin pretreatment. The thiobarbituric acid reactive substances (TBARS) level and superoxide dismutase (SOD) activity in the testes were evaluated. Subsequently, their spermatocytes were isolated, and DNA damage was assessed using an alkaline comet assay and the meiotic spread method. BPA administration did not significantly affect the weights of rats and their reproductive organs, and no alteration in sperm count was found. However, we demonstrated that BPA administration induced a significant increase in TBARS levels and a decrease in SOD activity that were concomitant with an increase in DNA migration within male germ cells and γH2AX foci formation on the autosomes of pachytene spermatocytes. Furthermore, a decrease in the proportion of 4C-cells was observed. These BPA effects were significantly alleviated by melatonin pretreatment. Nevertheless, the genotoxic effects of BPA were not accompanied by apoptosis in germ cells and morphological changes in the testes. These results indicate that BPA exposure may induce DNA damage accumulation in germ cells via oxidative stress. Moreover, melatonin may be a promising pharmacological candidate for preventing the potential genotoxicity of BPA following occupational or environmental exposure.

NF-kappaB is involved in inhibition of lipoxin A4 on dermal inflammation and hyperplasia induced by mezerein.

The mechanisms by which lipoxin A(4) (LXA(4)) inhibit skin inflammation remain unclear. In the present studies, the ear inflammatory model was induced by topical application of mezerein. Treatment of the mouse ear with LXA(4) exhibited the inhibitory effects on oedema, neutrophil infiltration, vascular permeability, expressions of interleukin (IL)-1, IL-6 and IL-8 mRNA, DNA-binding activity of nuclear factor-kappaB (NF-kappaB), and on dermal hyperplasia. NF-kappaB reporter activities and nuclear translocations of NF-kappaB p65 in cultured keratinocytes stimulated by mezerein were inhibited by pretreatment of the cells with LXA(4). LXA(4) reduced degradation, but not phosphorylation of IkappaBalpha in cultured keratinocytes stimulated by mezerein, suggesting that LXA(4)-attenuated IkappaBalpha degradation may restore the mezerein-blocked inhibitory effects of IkappaB on nuclear translocation and DNA-binding activity of NF-kappaB. Our results demonstrated that LXA(4) displays the anti-inflammatory and anti-proliferative role on ear inflammatory model induced by mezerein and these effects were related with downregulation of DNA-binding activity of NF-kappaB.