Duck pan-genome reveals two transposon insertions caused bodyweight enlarging and white plumage phenotype formation during evolution.

Structural variations (SVs) are a major source of domestication and improvement traits. We present the first duck pan-genome constructed using five genome assemblies capturing ∼40.98 Mb new sequences. This pan-genome together with high-depth sequencing data (∼46.5×) identified 101,041 SVs, of which substantial proportions were derived from transposable element (TE) activity. Many TE-derived SVs anchoring in a gene body or regulatory region are linked to duck's domestication and improvement. By combining quantitative genetics with molecular experiments, we, for the first time, unraveled a 6945 bp Gypsy insertion as a functional mutation of the major gene IGF2BP1 associated with duck bodyweight. This Gypsy insertion, to our knowledge, explains the largest effect on bodyweight among avian species (27.61% of phenotypic variation). In addition, we also examined another 6634 bp Gypsy insertion in MITF intron, which triggers a novel transcript of MITF, thereby contributing to the development of white plumage. Our findings highlight the importance of using a pan-genome as a reference in genomics studies and illuminate the impact of transposons in trait formation and livestock breeding.

Effects of residual feed intake on the economic traits of fast-growing meat ducks.

Feed efficiency (FE) is a crucial economic indicator of meat duck production. The objective of this study was to assess the impact of residual feed intake (RFI), defined as the difference between the actual and expected feed intake based on animal's production and maintenance requirements, on the growth performance (GP), slaughter and internal organ characteristics of fast-growing meat ducks. In total, 1,300 healthy 14-day-old male fast-growing meat ducks were housed in individual cages until slaughter at the age of 35 d. The characteristics of the carcass and internal organs of 30 ducks with the highest RFI (HRFI) and the lowest RFI (LRFI) were respectively determined. RFI, the feed conversion ratio (FCR), and average day feed intake (ADFI) were significantly lower in the LRFI group than the HRFI group (P < 0.001), while there were no significant differences in marketing BW or BW gain (BWG) (P > 0.05). The thigh muscle and lean meat yields were higher, and the abdominal fat content was lower (P < 0.001) in the LRFI group, while there were no significant differences in other carcass traits between the groups (P > 0.05). The liver and gizzard yields were significantly higher (P < 0.001) in the LRFI group, while there were no significant differences (P > 0.05) in intestinal length between the groups. RFI was highly positively correlate with FCR and ADFI (P < 0.01), but negatively correlated the yields of thigh muscle, lean meat, liver, and gizzard, and positively correlated with abdominal fat content. These results indicate that selection for low RFI could improve the FE of fast-growing meat ducks without affecting the marketing BW and BWG, while increasing yields of thigh muscle and lean meat and reducing abdominal fat content. These findings offer useful insights into the biological processes that influence FE of fast-growing meat ducks.

Mapping and Functional Dissection of the Rumpless Trait in Piao Chicken Identifies a Causal Loss of Function Mutation in the Novel Gene Rum.

Rumpless chickens exhibit an abnormality in their tail development. The genetics and biology of this trait has been studied for decades to illustrate a broad variation in both the types of inheritance and the severity in the developmental defects of the tail. In this study, we created a backcross pedigree by intercrossing Piao (rumpless) with Xianju (normal) to investigate the genetic mechanisms and molecular basis of the rumpless trait in Piao chicken. Through genome-wide association and linkage analyses, the candidate region was fine-mapped to 798.5 kb (chromosome 2: 86.9 to 87.7 Mb). Whole-genome sequencing analyses identified a single variant, a 4.2 kb deletion, which was completely associated with the rumpless phenotype. Explorations of the expression data identified a novel causative gene, Rum, that produced a long, intronless transcript across the deletion. The expression of Rum is embryo-specific, and it regulates the expression of MSGN1, a key factor in regulating T-box transcription factors required for mesoderm formation and differentiation. These results provide genetic and molecular experimental evidence for a novel mechanism regulating tail development in chicken and report the likely causal mutation for the tail abnormity in the Piao chicken. The novel regulatory gene, Rum, will, due to its role in fundamental embryo development, be of interest for further explorations of a potential role in tail and skeletal development also in other vertebrates.

Microstructure and Thermal Deformation Behavior of Hot-Pressing Sintered Zr-6Al-0.1B Alloy.

Zr-6Al-0.1B alloy rich in Zr3Al phase is prepared by hot-pressing sintering. The thermal deformation behavior of sintered Zr-6Al-0.1B is analyzed by isothermal compression tests at deformation temperatures of 950, 1050, and 1150 °C with strain rates of 0.01, 0.1, and 1 s-1. The results indicate that at the early stage of thermal deformation, the stress increases rapidly with the increase of strain and then reaches the peak value. Subsequently, the stress decreases with the increase of strain under the softening effect. On the whole, the true stress-strain curve shifts to the high stress area with the increase of strain rate or the decrease of deformation temperature, so the sintered Zr-6Al-0.1B alloy belongs to the temperature and strain rate sensitive material. For the microstructure evolution of sintered Zr-6Al-0.1B during the isothermal compression, the high strain rate can improve the grain refinement. However, because sintered Zr-6Al-0.1B is a low plastic material, too high strain rate will exceed the deformation capacity of the material, resulting in an increase in defects. The increase of deformation temperature also contributes to grain refinement, but when the temperature is too high, due to the decomposition of Zr3Al phase, the deformation coordination of the material decreases, leading to the increase of the probability of the occurrence of defects. This study verified the feasibility of hot-pressing sintering to prepare Zr-6Al-0.1B alloy rich in Zr3Al phase and laid the foundation of "hot-pressing sintering + canning hot-extrusion" process of Zr-6Al-0.1B alloy components.

Microenvironment-Triggered Degradable Hydrogel for Imaging Diagnosis and Combined Treatment of Intraocular Choroidal Melanoma.

Human choroidal melanoma (HCM) is one of the most common primary intraocular tumors and easily provokes liver metastases owing to the lack of sensitive and noninvasive therapeutic methods. Concerning the imaging diagnostics and therapeutic predicaments for choroidal melanoma, we designed microenvironment-triggered degradable hydrogels (RENP-ICG@PNIPAM:Dox-FA) based on ultrasmall (<5 nm) rare-earth nanoparticles (RENPs) with enhanced NIR-II luminescence. The ultrasmall diameter can significantly enhance the NIR-II luminescence performance of RENPs. RENPs were encapsulated by a dual-response PNIPAM hydrogel, which could release drug by responding to heat energy and glutathione under the tumor microenvironment. The in vitro/in vivo NIR-II imaging detection and antitumor activity were also compared systematically after different treatment conditions on ocular choroidal melanoma-1 cells and tumor-bearing mice, respectively. Besides, the degradability of the hydrogel composites under physiological conditions could be conducive to enhance the photothermal-chemotherapeutic effect and alleviate long-term biological toxicity. Our work on the microenvironment-triggered hydrogels with enhanced NIR imaging and easy metabolism may provide a promising strategy for sensitive and noninvasive imaging and phototherapy in ocular tumors.

Effects of Rearing Conditions and Sex on Cecal Microbiota in Ducks.

The intestinal microbiome influences the health of animals. However, little is known about the impact of indoor conditions and sex on intestinal microbiome diversity and composition in ducks. The present study aimed to investigate differences in the cecal microbiome between male and female ducks reared on the floor (PY group) or in cages (LY group). We also determined the relationships between cecal microbiota composition and slaughter traits, and the expression levels of mucosal and intestinal structural genes in ducks. There was a slight difference in slaughter traits among the groups, with cecum weight being significantly lighter in the LY compared with the PY group, especially in females (p < 0.05). Analysis of the alpha diversity of the cecal microbiota between males and females in the LY and PY groups showed that LY males had significantly lower diversity and richness. Beta diversity analysis demonstrated differences in the microbiota composition in relation to rearing conditions, and a significant difference between the sexes in the PY groups. The dominant bacterial phyla in duck cecum were Bacteroidetes, Firmicutes, Proteobacteria, and Fusobacteria. The relative abundances of the most common bacteria revealed that the intestinal microbiota diversity and composition were affected by both feeding conditions and sex. Several bacterial genera were detected differentially among the groups. These genera were correlated with slaughter traits and expression levels of mucosal and cecal structural genes in ducks. In conclusion, rearing conditions, sex, and associated changes in the cecal microbiota are thus associated with gut barrier functions in ducks.

Riemerella anatipestifer infection affects intestinal barrier structure and immune reactions in the duck caecum.

Riemerella anatipestifer (RA) infection causes high mortality and poor feed conversion, leading to great economic losses to the duck industry. This study investigated the effects of RA on the intestinal morphology and immune response of ducks. Histological examination showed that RA infection caused intestinal injury, including significantly reduced mucosal thickness on days 2, 3 and 5, significantly reduced villus height on days 1, 2, 3 and 5 (P < 0.05) and significantly reduced villus height to crypt depth ratios on days 2, 3, 5 and 9 of RA infection (P < 0.05). The expression of intestinal mucosal layer construction-associated genes and tight junction genes was significantly altered on at least one time point (day 1, 2, 3, 5, 9 or 14) after RA infection. Quantitative real-time polymerase chain reaction revealed that RA infection affected intestinal mucosal immune function. The genes encoding TLR4 (toll like receptor-4), TRAF6 (TNF receptor-associated factor 6), MYD88 (myeloid differentiation factor 88), IFN-γ (interferon-γ), IL (interleukin)-4 and IL-8 were significantly upregulated on day 2 of RA infection. Taken together, these results indicate that RA infection negatively affects intestinal barrier function in ducks due to impaired mucosal and villus-crypt structure and alters the mRNA expression of mucous layer construction-, intestinal tight junction-, and intestinal mucosal immunity-related genes.

Analysis of microbial diversity and composition in small intestine during different development times in ducks.

We studied the microbial profiles of the duodenum, jejunum, and ileum during different developmental stages in the duck using high-throughput sequencing of the bacterial 16S rRNA gene. We also investigated the differences in the microbiota in the duodenum, jejunum, and ileum at different developmental times. A correlation analysis was performed between the most abundant bacterial genera and the development of the small intestine. An analysis of alpha diversity indicated different species richness and bacterial diversity in the different small intestinal segments and at different development times. A beta diversity analysis indicated differences in the bacterial community compositions across time. In a weighted UniFrac principal coordinates analysis, the samples clustered into two categories, 2 to 4 wk and 6 to 10 wk, in the duodenum, jejunum, and ileum. Our results show that the small intestine is predominantly populated by the phyla Firmicutes, Bacteroidetes, and Proteobacteria throughout the developmental stages of the duck. The duodenum, jejunum, and ileum shared most of the bacterial phyla and genera present, although they showed significant differences in their relative abundances in the intestinal segments and developmental stages. They shared different bacterial taxa during development times and among different segments when the intergroup differences were analyzed. The genera Bacillus, Corynebacterium 1, Lactococcus, Sphingomonas, and Haliangium correlated moderately positively with the increase in bodyweight and the lengths and weights of the duodenum, jejunum, and ileum, and these genera may be considered important markers when assessing the heath of the intestinal microbiota in ducks. This study provides a foundation upon which to extend our knowledge of the diversity and composition of the duck microbiota and a basis for further studies of the management of the small intestinal microbiota and improvements in the health and production of ducks.

Hot Deformation Behavior of Q345 Steel and Its Application in Rapid Shear Connection.

The high-temperature deformation behavior of Q345 steel is detected by a Gleeble-3800 thermal simulator. The Arrhenius constitutive equation for high-temperature flow stress and the dynamic recrystallization model are constructed. With the secondary development technology, customized modifications are made on existing Deform-3D software. The constructed constitutive model and dynamic recrystallization model are embedded into Deform-3D to realize the secondary development of Deform-3D. The grain size and volume percentage distribution of dynamic recrystallization are obtained by simulating the shear connection process at high temperature and high speed. The results show that the constitutive equation and the dynamic recrystallization model constructed in this paper can be used to predict the evolution of the microstructure. The difference between the prediction results and the experimental data is about 3%. The accuracy of Arrhenius constitutive equation, dynamic recrystallization model and the feasibility of software secondary development are verified.

Effects of ammonia on intestinal microflora and productive performance of laying ducks.

Atmospheric ammonia is harmful to poultry and human health. The effect of ammonia on the intestinal microflora of laying ducks is still unknown. In this study, the effects of atmospheric ammonia and exposure time on the intestinal microflora of laying ducks were investigated using 16S rDNA sequencing technology. The body weight, ovary weight, spleen weight, liver weight, and productive performance of laying ducks were also recorded, and the relationship between intestinal microflora diversity and productive performance was analyzed. The results showed that Bacteroidetes, Firmicutes, and Proteobacteria were the dominant bacterial phyla. At the phylum and genus levels, with the exception of the phylum Firmicutes and the genus Sutterella, the top 10 most abundant phyla and genera differed significantly when the ammonia concentration was increased from 10 to 75 ppm and/or the exposure time was extended from 10 to 30 D. Laying rate was highly significantly lower in ducks exposed to 75 ppm ammonia for 10 D compared with those exposed to 10 ppm ammonia for 10 D. Body, ovary, and spleen weights also decreased when the ammonia concentration was increased. At the genus level, Flavonifractor was highly significantly positively correlated with ovary weight. Methanocorpusculum and Anaerotruncus were significantly positively correlated with ovary weight. Lactobacillus was significantly positively correlated with spleen weight. Phascolarctobacterium, Sphaerochaeta, Erysipelotrichaceae_UCG.004, and Lactococcus were significantly positively correlated with spleen weight. These results indicated that ammonia affected the diversity of the intestinal microbiota and the productive performance of laying ducks. Several intestinal microbiota genera were also correlated with organ weights.

Deep RNA sequencing of pectoralis muscle transcriptomes during late-term embryonic to neonatal development in indigenous Chinese duck breeds.

Pectoral muscle (PM) comprises an important component of overall meat mass in ducks. However, PM has shown arrested or even reduced growth during late embryonic development, and the molecular mechanisms underlying PM growth during the late embryonic to neonatal period in ducks have not been addressed. In this study, we characterized potential candidate genes and signaling pathways related to PM development using RNA sequencing of PM samples selected at embryonic days (E) 21 and 27 and 5 days post-hatch (dph) in two duck breeds (Gaoyou and Jinding ducks). A total of 393 differentially expressed genes (DEGs) were identified, which showed higher or lower expression levels at E27 compared with E21 and 5 dph, reflecting the pattern of PM growth rates. Among these, 43 DEGs were common to all three time points in both duck breeds. These DEGs may thus be involved in regulating this developmental process. Specifically, KEGG pathway analysis of the 393 DEGs showed that genes involved with different metabolism pathways were highly expressed, while genes involved with cell cycle pathways showed lower expression levels at E27. These DEGs may thus be involved in the mechanisms responsible for the phenomenon of static or decreased breast muscle growth in duck breeds during the late embryonic period. These results increase the available genetic information for ducks and provide valuable resources for analyzing the mechanisms underlying the process of PM development.

Erythropoietin facilitates the recruitment of bone marrow mesenchymal stem cells to sites of spinal cord injury.

Despite the successes of bone marrow mesenchymal stem cell (BMSC) transplantation for the treatment of spinal cord injuries, only a small fraction of grafted cells migrate to the target areas. Therefore, there remains a need for more efficient strategies of BMSC delivery. The present study was designed to explore this. Rat models of spinal cord injury (SCI) were established and exposed to phosphate buffered saline (control), BMSCs or BMSCs + erythropoietin (EPO). Basso, Beattie and Bresnahan (BBB) locomotor scale and grid walk tests were then utilized to estimate neurological rehabilitation. Additionally, the following assays were performed: Immunofluorescence localization of BMSCs to the site of SCI; the transwell migration assay to detect in vitro cellular migration; the terminal deoxynucleotidyl transferase dUTP nick end labeling assay to determine the apoptotic index of the lesion; and western blotting analysis to evaluate the expression of vascular endothelial growth factor (VEGF) and brain derived neurotrophic factor (BDNF) at the site of SCI. The BBB scores of the BMSC + EPO treated group were significantly increased compared with the BMSC treatment group (P<0.05). For example, BMSC + EPO treated rats had a significantly decreased number of hind limb slips compared with the BMSC treatment group (P<0.05). Furthermore, EPO significantly increased the migration capacity of BMSCs compared with the control group (P<0.001). In addition, the apoptotic index of the BMSC + EPO group was significantly decreased compared with the BMSC group (P<0.05). Green fluorescent protein-labeled BMSCs were detected at the site of SCI in the BMSC and BMSCs + EPO groups, with the signal being notably stronger in the latter. Moreover, the expression of VEGF and BDNF in the BMSCs + EPO group was significantly increased compared with the BMSC group (P<0.05). In conclusion, the results of the present study indicate that EPO can facilitate the recruitment of BMSCs to sites of SCI, increase expression of BDNF and VEGF, and accelerate recovery of neurological function following SCI.

Episodes of HIV viremia and the risk of non-AIDS diseases in patients on suppressive antiretroviral therapy.

BACKGROUND: Several studies reported an association between immunodeficiency and non-AIDS-defining diseases. We investigated whether nonstructured treatment interruptions and episodes of viremia during suppressive combination antiretroviral therapy were independently associated with non-AIDS diseases. METHODS: Six thousand four hundred forty patients with viral suppression (<50 copies/mL) within 48 weeks of starting combination antiretroviral therapy were selected from the Dutch ATHENA cohort. In proportional hazards models, associations between treatment interruptions, viral suppression, low-level (50-400 copies/mL), and high-level viremia (>400), and serious non-AIDS diseases (cardiovascular disease, chronic renal failure, liver fibrosis/cirrhosis) were investigated by including time-updated cumulative exposure to either viremia and interruptions or HIV RNA >400 copies per milliliter. RESULTS: During 24,603 person-years, of which 88.5% occurred during viral suppression, 102 patients developed cardiovascular disease, 54 chronic renal failure, and 70 liver fibrosis/cirrhosis. Overall incidence of non-AIDS diseases ranged from 1.41 (95% confidence interval: 0.73 to 2.46) per 100 person-years for CD4 counts <200 to 0.71 (0.49 to 1.00) for CD4 ≥500 cells per cubic millimeter. Compared with viral suppression, high-level viremia was associated only with cardiovascular disease (relative hazard: 1.37, 1.04 to 1.81 per year longer), whereas interruptions and low-level viremia were not associated with non-AIDS diseases. Relative hazards for cumulative exposure to RNA >400 versus ≤400 copies per milliliter were 1.32 (1.01 to 1.73) for cardiovascular disease, 1.13 (0.66 to 1.92) for renal failure, and 0.86 (0.51 to 1.44) for fibrosis/cirrhosis. CONCLUSIONS: Lower CD4 counts are associated with increased risk of non-AIDS diseases, whereas high-level viremia seems to be independently associated with cardiovascular disease. However, the power to detect associations with viremia or interruptions may have been limited as most events occurred during viral suppression.

Long-term complications in patients with poor immunological recovery despite virological successful HAART in Dutch ATHENA cohort.

OBJECTIVE: We investigated the risk of AIDS and serious non-AIDS-defining diseases (non-ADDs) according to the degree of immunological recovery after 2 years of virological successful antiretroviral therapy (HAART). DESIGN: Retrospective observational cohort study including HIV-infected patients treated with HAART resulting in viral suppression (<500 copies/ml). METHODS: Patients were grouped according to their CD4 cell count after 2 years of HAART: CD4 cell count less than 200 cells/µl (group A), 200-350 cells/µl (group B), 351-500 cells/µl (group C) or more than 500 cells/µl (group D). Analysis was done to assess predictors for poor immunological recovery and the occurrence of a composite endpoint [death, AIDS, malignancies, liver cirrhosis and cardiovascular events (CVEs)], non-ADDs, CVEs and non-AIDS-defining malignancies (non-ADMs). RESULTS: Three thousand and sixty-eight patients were included. Older age, lower CD4 cell nadir and lower plasma HIV-RNA at the start of HAART were independent predictors for a poor immunological recovery. The composite endpoint, non-ADDs and CVE were observed most frequently in group A (overall log rank, P < 0.0001, P = 0.002 and P = 0.01). In adjusted analyses, age was a strong independent predictor for all endpoints. Compared with group A, patients in group D had a lower risk for the composite endpoint [hazard ratio 0.54 (95% confidence interval [CI] 0.33-0.87]; patients in group B had a lower risk for CVEs [hazard ratio 0.34 (95% CI 0.14-0.86)]. CONCLUSION: Poor immunological recovery despite virological successful HAART is associated with a higher risk for overall morbidity and mortality and CVEs in particular. This study underlines the importance of starting HAART at higher CD4 cell counts, particularly in older patients.

Prognostic value of Her-2/neu and clinicopathologic factors for evaluating progression and disease-specific death in Chinese men with prostate cancer.

BACKGROUND: Her-2/neu gene overexpression has been found in several malignancies, and is associated with poor prognosis; while its role in the tumorigenesis and progression of prostate cancer (PCa) is still controversial. This study aimed to evaluate the prognostic value of Her-2/neu protein expression and clinicopathologic factors in antiandrogen-treated Chinese men with PCa for disease progression and PCa-specific death. METHODS: Her-2/neu protein expression was determined using immunohistochemistry (IHC) in specimens collected from 124 prostate biopsies and transurethral resection of prostate (TURP) from seven prostate cancer patients. RESULTS: Her-2/neu protein expression was 0, 1+, 2+, and 3+ in 40 (30.5%), 8 (6.1%), 67 (51.1%), and 16 (12.2%) cases, respectively. Her-2/neu protein expression showed significant correlation as judged by Gleason score (P = 0.049), clinical tumor-node-metastases (cTNM) stage (P = 0.018) and disease progression (P = 0.001), but did not correlate with prostate-specific antigen (PSA) (P = 0.126) or PCa-specific death (P = 0.585). PSA (P = 0.001), Gleason score (P = 0.017), cTNM (P = 0.000) and Her-2/neu protein expression (P = 0.001) had prognostic value for evaluating the progression of PCa in univariate analysis. In Kaplan-Meier plots, both Gleason score (P = 0.035) and cTNM (P = 0.013) correlated with PCa-specific death. In multivariate analysis, only cTNM was significant for both disease progression (P = 0.001) and PCa-specific death (P = 0.031). CONCLUSIONS: Her-2/neu protein expression is significantly correlated with Gleason score, cTNM and disease progression, although it is not an independent predictor of disease progression and PCa-specific death. cTNM staging serves as an independent prognostic factor for disease progression and PCa-specific death.

Clinical significance of transient HIV type-1 viraemia and treatment interruptions during suppressive antiretroviral treatment.

BACKGROUND: Transient episodes of HIV type-1 viraemia are frequently observed in patients on suppressive combination antiretroviral therapy (cART). We studied the effect of such episodes and of treatment interruptions on clinical outcome and immunological response. METHODS: A total of 3,321 patients from the ATHENA cohort had virological suppression (HIV type-1 RNA<50 copies/ml) after 24 weeks of cART. The association between subsequent episodes of treatment interruptions, viral suppression, low-level (50-400 copies/ml) and high-level (>400 copies/ml) viraemia and the outcomes death, AIDS or immunological response (CD4(+) T-cell count increase > or =50% from 24 weeks) was studied with Poisson regression models, including either time-updated cumulative follow-up, time spent per type of episode or modelling episodes as binary status indicators. RESULTS: During 11,165 person-years of follow-up, 88 patients died, 111 developed AIDS and 2,019 had an immunological response. Longer follow-up time in treatment interruptions increased the risk of AIDS (relative risk [RR] 8.07, 95% confidence interval [CI] 3.98-16.4 per year longer) and impaired immunological response (RR 0.22, 95% CI 0.12-0.41). High-level viraemia was only associated with immunological response (RR 0.55, 95% CI 0.40-0.74), whereas low-level viraemia was not associated with any of the three outcomes. Status indicator models gave similar results. When also including time-updated CD4(+) T-cell counts, the observed associations diminished. CONCLUSIONS: Treatment interruptions and high-level, but not low-level, viraemia are strongly associated with clinical outcome, mainly via their effect on CD4(+) T-cell counts.

Involvement of 90-kuD ribosomal S6 kinase in collagen type I expression in rat hepatic fibrosis.

AIM: To investigate the relationship between 90-kuD ribosomal S6 kinase (p90RSK) and collagen type I expression during the development of hepatic fibrosis in vivo and in vitro. METHODS: Rat hepatic fibrosis was induced by intraperitoneal injection of dimethylnitrosamine. The protein expression and cell location of p90RSK and their relationship with collagen type I were determined by co-immunofluoresence and confocal microscopy. Subsequently, RNAi strategy was employed to silence p90RSK mRNA expression in HSC-T6, an activated hepatic stellate cell (HSC) line. The expression of collagen type I in HSC-T6 cells was assessed by Western blotting and real-time polymerase chain reaction. Furthermore, HSCs were transfected with expression vectors or RNAi constructs of p90RSK to increase or decrease the p90RSK expression, then collagen type I promoter activity in the transfected HSCs was examined by reporter assay. Lastly HSC-T6 cells transfected with p90RSK siRNA was treated with or without platelet-derived growth factor (PDGF)-BB at a final concentration of 20 microg/L and the cell growth was determined by MTS conversion. RESULTS: In fibrotic liver tissues, p90RSK was over-expressed in activated HSCs and had a significant positive correlation with collagen type I levels. In HSC-T6 cells transfected with RNAi targeted to p90RSK, the expression of collagen type I was down-regulated (61.8% in mRNA, P < 0.01, 89.1% in protein, P < 0.01). However, collagen type I promoter activity was not increased with over-expression of p90RSK and not decreased with low expression either, compared with controls in the same cell line (P = 0.076). Furthermore, p90RSK siRNA exerted the inhibition of HSC proliferation, and also abolished the effect of PDGF on the HSC proliferation. CONCLUSION: p90RSK is over-expressed in activated HSCs and involved in regulating the abnormal expression of collagen type I through initiating the proliferation of HSCs.

Immunologic, virologic, and clinical consequences of episodes of transient viremia during suppressive combination antiretroviral therapy.

OBJECTIVE: To investigate immunologic, virologic, and clinical consequences of episodes of transient viremia in patients with sustained virologic suppression. METHODS: From the AIDS Therapy Evaluation Project, Netherlands cohort, 4447 previously therapy-naive patients were selected who were on continuous combination antiretroviral therapy and had initial success (2 consecutive HIV RNA measurements <50 copies/mL). During episodes of viral suppression (RNA <50 copies/mL), low-level viremia (RNA 50 to 1000 copies/mL), or high-level viremia (RNA >1000 copies/mL) after initial success, the occurrence of therapy changes, drug resistance, and clinical events was assessed. RESULTS: During 11,187 person-years of follow-up, 1281 (28.8%) patients had at least 1 RNA measurement >50 copies/mL. Among 8069 episodes, there were 5989 (74.2%) episodes of suppression, 1711 (21.2%) episodes of low-level viremia, and 369 (4.6%) episodes of high-level viremia. Most episodes of low-level viremia consisted of < or =2 RNA measurements (93.7%), were without clinical events or therapy changes (79.6%), and were without changes in CD4 cell counts. Therapy changes (52.3% of episodes) and resistance (23.3%) were frequently observed during high-level viremia. CONCLUSIONS: Episodes of low-level viremia are frequent and short lasting, and the low proportion of episodes with clinical events suggests that leaving therapy unchanged is a clinically acceptable strategy. In contrast, high-level viremia is associated with resistance and is often followed by therapy changes.

The effects of isosteviol against myocardium injury induced by ischaemia-reperfusion in the isolated guinea pig heart.

1. This study aimed to investigate the protective effects of isosteviol against myocardial ischaemia-reperfusion (IR) injury and its effects on mitochondrial adenosine triphosphate (ATP)-sensitive potassium channel (mitoK(ATP)) activity in vitro. 2. Groups of eight guinea pigs were treated as follows: constant perfusion control (PC), IR control, ischaemic preconditioning (IPC) + IR, isosteviol (50, 250 or 500 nmol) + IR, 5-hydroxydecanoate acid (5-HD) (5 micromol) + isosteviol (500 nmol) + IR. The guinea pig heart was isolated and perfused in Langendorff mode with modified Tyrode solution at a flow rate of 10 mL/min. Ischaemia was introduced for 30 min followed by reperfusion for 20 min. Cardiac function, coronary arterial flow rate, lactate dehydrogenase (LDH) and creatine kinase (CK) activities in the perfusate were measured prior to ischaemia and at the end of reperfusion. 3. There were no significant (P > 0.05) changes in cardiac function or markers of cell damage (i.e. activities of LDH and CK) in the PC group. In contrast, cardiac function was adversely affected in the IR group, with significant (P < 0.05) decreases in left ventricular developing pressure (LVDevP), dP/dt(max) and dP/dt(min) compared with baseline and the PC group. In addition, there were increases in activity of LDH (20%) and CK (67%) compared with baseline and the PC group. 4. Ischaemic preconditioning and pretreatment with isosteviol, at all dose levels, resulted in a significant (P < 0.05) attenuation of IR injury. Lactate dehydrogenase and CK activities were not significantly (P < 0.05) different compared with baseline. Isosteviol did not increase coronary flow, suggesting that the protective effect of isosteviol on the myocardium was not mediated by dilation of the coronary blood vessels. 5. Pretreatment with the mitoK(ATP) blocker 5-HD partially antagonized the effects of 500 nmol isosteviol, with a statistically significant attenuation of its protective effects on HR, LVDevP, dP/dt(max) and dP/dt(min) compared with isosteviol alone pretreatment. 6. The IR injury on the Langendorff perfused guinea pig heart was alleviated by isosteviol, which appears to mediate its effects through mitoK(ATP) channels. Future research might aim to investigate the interaction of isosteviol with mitoK(ATP) channels in order to clarify its mechanism of action.

The cardioprotective effect of isosteviol on rats with heart ischemia-reperfusion injury.

This study was designed to assess the cardioprotective effect of isosteviol on rats with heart ischemia-reperfusion (IR) injury and to explore the mechanism of action of the compound. Sprague Dawley rats were divided into 8 groups (n=10-12): a sham-operated control and 7 ischemia-reperfusion groups (IR control, 3 isosteviol pre-treated (0.5, 1.0 and 2.0 mg kg(-1)), ligustrazine pre-treated, 5-hydroxydecanoate (5-HD) pre-treated and 5-HD+ isosteviol pre-treated groups). IR was produced by occluding the left coronary artery for 30 min followed by re-opening the artery for 90 min. The compounds under investigation were administered intravenously 10 min prior to occluding the artery. Hemodynamic parameters (+/-dp/dt(max), LVSP, LVDevP, MAP), heart rate, ventricular tachycardia (VT) and ventricular fibrillation (VF) were determined during the IR period. The myocardial infarct size, activities of serum lactate dehydrogenase and creatine kinase were determined at the end of the experiment. In the isosteviol pre-treated groups, the hemodynamic parameters were improved and the myocardial infarct size, the activities of serum enzymes, and the incidences of VT and VF were all decreased when compared to the control group. These effects of isosteviol were similar to that of a traditional cardioprotective agent, ligustrazine. The 5-HD+ isosteviol group displayed parameters that were between those in the equivalent isosteviol pre-treated group and the IR control group. In conclusion, damage due to a standard rat heart IR injury was reduced by pretreatment with intravenous isosteviol, and this effect was partly attenuated by a mitochondrial ATP-sensitive potassium channel blocker, 5-HD.