Amorphophallus konjac anthracnose caused by Colletotrichum siamense in China.

AIMS: Amorphophallus konjac is an important commercial crop grown in China because it is the only plant species which is rich in glucomannan concentration. Recently, an outbreak of anthracnose (incidence ranging from 10-15%) was observed in a field survey conducted from June to August 2018. This study aims to identify the causal agent of A. konjac anthracnose. METHODS AND RESULTS: The pathogen was isolated on potato dextrose agar (PDA) medium. The fungal colony on PDA was greyish to dark grey. Conidia were falcate, one-celled and hyaline. Based on the micro-morphological and cultural characteristics, the pathogen was identified as Colletotrichum sp. blast search and phylogenetic analysis of the ITS, GAPDH, CHS1, ACT, CAL and TUB2 genes revealed the pathogen as Colletotrichum siamense. Koch's postulates were conducted on 2-month konjac leaves with conidial suspension. Development of typical anthracnose disease was recorded 5 days after inoculation and the pathogen's identity was confirmed by re-isolation and molecular identification. CONCLUSIONS: Amorphophallus konjac anthracnose was caused by C. siamense in China. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of causal agent of A. konjac anthracnose will be helpful in designing effective disease control strategies.

Application of confocal, SHG and atomic force microscopy for characterizing the structure of the most superficial layer of articular cartilage.

The surface of articular cartilage plays a crucial role in attenuating and transmitting mechanical loads in synovial joints to facilitate painless locomotion. Disruption to the surface of articular cartilage causes changes to its frictional properties instigating the deterioration of the tissue. In this study, we physically peeled the most superficial layer, a transparent membrane of 20.0 ± 4.7 µm thick, from the central loading region of femoral condyles of sheep. The ultrastructure of this layer without interference from the underlying cartilage was independently investigated using confocal, second harmonic generation and atomic force microscopy. We found that the most superficial layer contains chondrocytes, densely packed collagen, coarse elastic fibres and a fine elastic network. The elastic fibres are most prevalent at the surface of the layer, where collagen and chondrocyte densities are lowest. At the interface of this most superficial layer with the underlying bulk cartilage, a dense fibrillar network exists, formed mainly by collagen fibrils and elastin microfibrils. By contrast, the interface of the underlying cartilage with the most superficial layer contains collagen fibrils, fine microfibrils and microfibrils distinctively laced on one side. The findings of this study will play an important role in understanding the mechanical function and wear resistance of articular cartilage, and in developing more promising tissue engineering techniques to treat cartilage defects and osteoarthritis. LAY DESCRIPTION: The chronic pain and dysfuction in synovial joints caused by osteoarthritis can have a debilitating impact on daily activities for sufferers. Osteoarthritis is characterised by the deterioration of the articular cartilage. Despite intensive research, the wear mechanism of articular cartilage and the progression of osteoarthritis remain unclear in the literature. Articular cartilage is a resilient tissue that provides a low friction surface to facilitate painless locomotion. The surface of articular cartilage plays a crucial role in attenuating and transmitting mechanical loads. Disruption at the surface of articular cartilage causes changes to its frictional properties, instigating the deterioration of the tissue. Despite this, the definition of the most superficial layer of articular cartilage, as well as its composition and microstructure, have endured a long history of debate, clouding our understanding of the early progression of osteoarthritis. In order to investigate the surface of articular cartilage independently from the underlying cartilage, we physically peeled a transparent membrane of 20.0 ± 4.7 µm thickness, the most superficial layer, from the central loading region of the femoral condyles of sheep. Using confocal, second harmonic generation and atomic force microscopy, we found that the most superficial layer contains cartilage cells (chondrocytes), densely packed collagen, coarse elastic fibres and a fine elastic network. The coarse elastic fibres are most prevalent at the surface of the layer where collagen and chondrocyte densities are lowest. Furthermore, we investigated the surfaces at the interface of the most superficial layer with the underlying articular cartilage. At the interface of this most superficial layer with the underlying bulk cartilage, a dense fibrillar network exists, formed mainly by collagen fibrils and elastin microfibrils. In contrast, the interface of the underlying cartilage with the most superficial layer contains collagen fibrils, fine microfibrils and microfibrils distinctively laced on one side. The findings of this study have confirmed that there is a most superficial layer that is able to be removed using a tangential force. Through the application of advanced imaging technologies, we have shown that this most superficial layer is cellular and have detailed its composition and ultrastructure. Due to the close association between the form and function of tissues, the findings of this study will play an important role in understanding the mechanical function and wear mechanism of articular cartilage. This may lead to the development of more promising tissue engineering techniques to treat cartilage defects and osteoarthritis.

[Evaluation of carotid plaque neovascularization in carotid stenosis by contrast-enhanced ultrasound].

Objective: The identification of neovascularization in carotid plaque in carotid artery stenosis by contrast-enhanced ultrasound (CEUS) provides other risk markers for stroke besides carotid artery stenosis -intraplaque neovascularization. Methods: From January 2017 to September 2017, 40 patients with carotid atherosclerosis plaque were examined by contrast-enhanced ultrasound in China-Japanese Friendship Hospital. The enhancement intensity (EI) measured by contrast-enhanced ultrasound was compared with the micro-vessel density (MVD) measured by histopathology after carotid endarterectomy (CEA). Contrast-enhanced ultrasound was used to observe whether there was enhancement in the plaque and the enhancement was divided into 0-2 grades. The EI in plaque, the ratio of EI in plaque to EI in carotid artery lumen were calculated by time intensity curve quantitative analysis software. Pathological sections of carotid plaques after CEA were stained with CD34 and neovascularization density was measured. Results: There were significant differences in age, EI1, EI1/EI2 and CD34 among patients with different grades of plaque enhancement (P<0.05), but no significant differences in gender and EI2 (P>0.05). The density of neovascularization obtained by CD34 staining was highly positively correlated with EI1 (r=0.836, P<0.001), EI1/EI2 (r=0.955, P<0.001), but not with age (r=0.066, P=0.684), EI2 (r=0.159, P=0.328). Conclusions: Contrast-enhanced ultrasound can observe the neovascularization in carotid plaque, which is a simple and non-invasive method to evaluate the stability of carotid plaque. CEUS may also help to extract features of vulnerable plaques, such as acute intraplaque hemorrhage.

The association between murine double minute 2 (MDM2) rs2279744 and endometrial cancer risk in a Chinese Han population.

Some studies investigated the association between the murine double minute 2 (MDM2) rs2279744 polymorphism and endometrial cancer susceptibility, but provided controversial or inconclusive results. Thus, we decided to perform this case-control study to determine the association between MDM2 rs2279744 polymorphism and endometrial cancer in a Chinese Han population. A total of 215 endometrial cancer patients and 212 cancer-free controls were included in this case-control study. We genotyped the MDM2 rs2279744 polymorphism by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). GG genotype showed a statistically significantly increased risk of developing endometrial cancer (OR=1.72, 95%CI 1.08-2.76, P=0.02). Statistically significant difference was observed when the patients and controls were compared according to G versus I (OR=1.40, 95%CI 1.07-1.84, P=0.01). A significantly higher frequency of G allele was observed in patients with stage III+IV, compared to stage I+II (OR=2.24, 95%CI 1.49-3.38, P=0.001). In conclusion, the study found that MDM2 rs2279744 polymorphism was significantly associated with endometrial cancer risk in a Chinese Han population.

Polymorphisms of Flanking Region of the ASB15 Gene and Their Associations with Performance Traits in Chicken.

Research on the identity of genes and their relationship with traits of economic importance in chickens could assist in the selection of poultry. In this study, an F2 resource population of Gushi chickens crossed with Anka broilers was used to detect single-nucleotide polymorphisms (SNPs) in the flanking region of the ASB15 gene by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). One SNP of -1271 C>T in 5' flanking region of the chicken ASB15 gene and two SNPs of the 10618 A>G and 10716 G>A in 3' flanking region were identified. Furthermore, the 10618 A>G and 10716 G>A in 3' flanking region were in complete linkage. Association analysis results showed that -1271 C>T was not associated with performance traits, while the 10618 A>G and 10716 G>A were significantly associated with BW2, 4, 6, 8, 10, 12, SL12, CD8, CW4, 8, 12, BSL4, 8, 12, and SEW, EW, WW, BMW, LW, CW, SFT. Our results suggest that the ASB15 gene profoundly affects chicken performance traits.

Promoter analysis and tissue expression of the chicken ASB15 gene.

1. This study was conducted to explore the promoter region of the chicken ASB15 gene by detecting the activities of the dual luciferase reporter gene and to assess expression profiles of the ASB15 gene in 10 different tissues from Gushi chickens. 2. Five dual luciferase reporter gene vectors were constructed and transfected into DF1 cells. The activities of recombined plasmids were measured and the core promoter was confirmed by bioinformatic analysis. Total RNA was extracted and the relative expression of the ASB15 gene was examined. 3. Data analysis indicated that the promoter was located from -955 to -212 bp. Results showed that the chicken ASB15 gene was expressed in heart, breast muscle and leg muscle. 4. This study has confirmed the promoter region and the expression profile of the chicken ASB15 gene, which provides a foundation for further exploring its transcriptional regulation and function.

Comparative analysis of the liver tissue transcriptomes of Mongolian and Lanzhou fat-tailed sheep.

Research on gene regulation has been made possible with the help of RNA sequencing applications such as RNA-Seq technology for high-throughput sequencing platforms. Recent studies have explored the transcriptomes from different tissues of domestic animals using RNA-Seq technology, but little research has been done to study the transcriptomes of breeds of sheep having different adipose tissue deposition mechanisms, such as Mongolian and Lanzhou fat-tailed sheep. In this study, Mongolian and Lanzhou fat-tailed sheep were selected as experimental breeds, and six libraries (three libraries per breed) were constructed. A total of 286 Mb of high-quality reads was obtained, and three-quarters of the reads were mapped to the reference genome per library. In addition, there were 16,257, 16,186, 16,254, 16,827, 16,437, and 15,761 known reference genes in the six constructed libraries (LCL1, LCL2, LCL3, MCL1, MCL2, and MCL3, respectively). Seven genes were differentially expressed: four were upregulated and three were downregulated in liver tissue between the MCL and LCL groups; 65,303, 65,442, 63,426, 76,267, 69,853, and 57,439 potential cSNPs were detected in the six libraries, respectively, with the G/R substitution occurring most commonly. There were 24,239, 22,283, 22,457, 26,635, 27,093, and 18,700 alternate splicing (AS) events in the six libraries. Intron retention was the most common AS event, followed by alternative 3' splice sites. These results indicate that there are many differences in the liver transcriptomes of Mongolian and Lanzhou fat-tailed sheep breeds. Such results may provide fundamental information for further research on defining the sheep genome.

Effects of tail docking on the expression of genes related to lipid metabolism in Lanzhou fat-tailed sheep.

To evaluate stearoyl-CoA desaturase (SCD), hormone-sensitive lipase (HSL), lipoprotein lipase (LPL), and peroxisome proliferator-activated receptor (PPARγ) expression in Lanzhou fat-tailed sheep (with and without docked tails), 18 rams were randomly divided into two equal groups (docked group, LT; control group, LC). These data were also used to increase the understanding of sheep fat deposition and metabolism. All animals were harvested at the age of 18 months, and expression was determined for 10 tissues. The results indicated that the fat weight of each tissue in LT was higher than in LC (P < 0.05). SCD expression in semitendinosus, omentum majus fat (OF), subcutaneous fat, kidney fat (KF), and subcutaneous rump fat was higher in LT than in LC rams (P < 0.05). Trends (P < 0.10) associated with higher HSL expression of LC in comparison to that of LT rams in intestinal fat, OF, and KF tissues were detected. Numerically, LPL expression was the highest in KF, OF, and kidney tissues, but there were few differences (P > 0.10). PPARγexpression was greater in LT than in LC rams in liver tissues (P < 0.05), but there were few differences in other tissues. No significant differences were found with regard to the regression analysis of expression and adipose tissue weights, but the two indices exhibited the same trend. The results indicated that changes in fatty deposits may be due to the common control of docking management and the minor effects associated with the regulation of SCD, HSL, LPL, and PPARγexpression.

[The value of PHI/PCA3 in the early diagnosis of prostate cancer].

OBJECTIVE: To investigate the value of prostate health index (PHI) and prostate cancer gene 3 (PCA3) in the early diagnosis of prostate cancer (PCa). METHODS: A total of 190 patients with abnormal serum prostate specific antigen (PSA) or abnormal digital rectal examination were enrolled. They were all underwent initial biopsy and 11 of them were also underwent repeated biopsy. In addition, 25 healthy cases (with normal digital rectal examination and PSA<4 ng/ml) were the control group.The PHI and PCA3 were detected by using immunofluorescence and Loop-Mediated Isothermal Amplification (LAMP). The sensitivity and specificity of diagnosis were determined by ROC curve.In addition, the relationship between PHI/PSA and the Gleason score and clinical stage were analyzed. RESULTS: A total of 89 patients were confirmed PCa by Pathological diagnosis. The other 101 patients were diagnosed as benign prostatic hyperplasia (BPH). The sensitivity and specificity of PCA3 test were 85.4% was 92.1%. Area under curve (AUC) of PHI is higher than AUC of PSA (0.727>0.699). The PHI in peripheral blood was positively correlated with Gleason score and clinical stage. CONCLUSIONS: The detection of PCA3 and PHI shows excellent detecting effectiveness. Compared with single PSA, the combined detection of PHI and PCA3 improved the diagnostic specificity. It can provide a new method for the early diagnosis in prostate cancer and avoid unnecessary biopsies.

The development of confocal arthroscopy as optical histology for rotator cuff tendinopathy.

MRI, ultrasound and video arthroscopy are traditional imaging technologies for noninvasive or minimal invasive assessment of the rotator cuff tendon pathology. However, these imaging modalities do not have sufficient resolution to demonstrate the pathology of rotator cuff tendons at a microstructural level. Therefore, they are insensitive to low-level tendon diseases. Although traditional histology can be used to analyze the physiology of rotator cuff tendons, it requires biopsy that traumatizes the rotator cuff, thus, potentially comprising the mechanical properties of tendons. Besides, it cannot offer real-time histological information. Confocal endoscopy offers a way to assess the microstructural disorder in tissues without biopsy. However, the application of this useful technique for detecting low-level tendon diseases has been restricted by using clinical grade fluorescent contrast agent to acquire high-resolution microstructural images of tendons. In this study, using a clinical grade sodium fluorescein contrast agent, we have reported the development of confocal arthroscopy for optical histological assessment without biopsy. The confocal arthroscopic technique was able to demonstrate rotator cuff tendinopathy in human cadavers, which appeared macroscopically normal under video arthroscopic examinations. The tendinopathy status of the rotator cuff tendons was confirmed by corresponding traditional histology. The development of confocal arthroscopy may provide a minimally invasive imaging technique for real-time histology of rotator cuff without the need for tissue biopsy. This technique has the potential for surgeons to gain in real time the histological information of rotator cuff tendons, which may assist planning repair strategies and potentially improve intervention outcomes.

Estrogen deficiency inducing shifted cytokines profile in bone marrow stromal cells inhibits Treg cells function in OVX mice.

The study aims to reveal the effect of estrogen deficiency on Treg cells population in bone marrow in the development of osteoclastogenis with comparing the differences about Treg cells phenotypes and cytokines related with the homeostasis and functions maintenance of Treg cells in bone marrow in OVX mice and health control group. Wide—type C57BL/6 mice were operated OVX to mimic estrogen deficiency in PMO women. Treg cells population and their surface markers expressions were detected by flow cytometry. Cytokines profiles in bone marrow with examined by real—time PCR and ELISA analysis. Signal pathways and key modulators responsible to inflammatory cytokines expressions in bone marrow stromal cells were also detected with using western blotting. Estrogen deficiency in OVX mice decreased Treg cells and their functions, and cytokines profile in bone marrow were found shifted in bone marrow when compared with control group. Consistent to these observations, signal pathways in bone marrow stromal cells were reported altered by estrogen deficiency in our model. Estrogen deficiency effects Treg cells population and their functions in OVX mice with altering cytokines profile in bone marrow stromal cells.

Identification of single nucleotide polymorphisms in the ASB15 gene and their associations with chicken growth and carcass traits.

ASB15 is a member of the ankyrin repeat and suppressor of cytokine signaling box family, and is predominantly expressed in skeletal muscle. In the present study, an F2 resource population of Gushi chickens crossed with Anka broilers was used to investigate the genetic effects of the chicken ASB15 gene. Two single nucleotide polymorphisms (SNPs) (rs315759231 A>G and rs312619270 T>C) were identified in exon 7 of the ASB15 gene using forced chain reaction-restriction fragment length polymorphism and DNA sequencing. One was a missense SNP (rs315759231 A>G) and the other was a synonymous SNP (rs312619270 T>C). The rs315759231 A>G polymorphism was significantly associated with body weight at birth, 12-week body slanting length, semi-evisceration weight, evisceration weight, leg muscle weight, and carcass weight (P < 0.05). The rs312619270 T>C polymorphism was significantly associated with body weight at birth, 4, 8, and 12-week body weight, 8-week shank length, 12-week breast bone length, 8 and 12-week body slanting length, breast muscle weight, and carcass weight (P < 0.05). Our results suggest that the ASB15 gene profoundly affects chicken growth and carcass traits.

Fatty acid composition differences between adipose depot sites in dairy and beef steer breeds.

The objective of the study was to compare fatty acid composition of longissimus dorsi (LD) and kidney fat (KF) in Holstein steers (HS), Simmental steers (SS) and Chinese LongDong Yellow Cattle steers (CLD). All steers received the same nutrition and management but in different locations. Cattle were harvested at approximately 550 kg and fatty acid composition of longissimus dorsi and kidney fat was analyzed in samples taken after 3 days of aging. There was evidence (P < 0.05) that C18:3n6 was greater in KF than LD in CLD cattle but not in HS or SS cattle. Percentage C18:1n9, C18:2n6, C18:3n3, and n6 fatty acids were greater in LD than KF for all breeds (P < 0.05), but the difference between fat sources for n6 in CLD cattle was smaller than the other two breeds. The LD had greater percentage of polyunsaturated fatty acids (PUFA), monounsaturated fatty acids (MUFA), and a greater ratio of n6:n3 PUFAs compared to the KF in each breed (P < 0.05). The △(9)-desaturase catalytic activity index was greater in LD than in KF in each breed group (P < 0.05). Percentage cis-9, trans-11 CLA was greater in KF than LD in HS (P < 0.05) but not SS or CLD cattle. These results indicate fatty acid percentages generally differed between longissimus dorsi fat and kidney fat. Further, there was some indication that some of these differences between fatty acid deposition sites were not consistent across breed group.

Study of the correlation between GH gene polymorphism and growth traits in sheep.

The growth hormone gene plays an important role in the physiological function of an organism. The current study aimed to investigate the correlation between polymorphisms in the 5' regulatory region, exon 4, and 3' untranslated region (UTR) of the sheep GH gene and sheep growth traits. The DNA from 510 adult sheep was analyzed by DNA sequencing and polymerase chain reaction single-strand conformation polymorphism. Two alleles (A and B) and 3 genotypes (AA, AB, and BB), 2 alleles (A and B) and 3 genotypes (AA, AB, and BB), and 3 alleles (A, B, and C) and 4 genotypes (AA, AB, BB, and AC) were found within the 5' regulatory region, exon 4, and 3' UTR, respectively. In Tibetan sheep, the association analysis indicated that there were statistically significant differences in the scores of weight, length, and heart girth within the 5' regulatory region; weight, length, wither height, and heart girth within exon 4; and weight, length, wither height, and heart girth within the 3' UTR among the different genotypes. For exon 4, Poll Dorset sheep individuals with genotype AA showed a lower score than those of genotypes BB and AB (P<0.05). With regard to the 3' UTR, Poll Dorset sheep with genotype AC showed higher scores than those of genotypes AA and AB (P<0.05).

Establishment of the optimum two-dimensional electrophoresis system of ovine ovarian tissue.

Lambing performance of sheep is the most important economic trait and is regarded as a critic factoring affecting the productivity in sheep industry. Ovary plays the most roles in lambing trait. To establish the optimum two-dimensional electrophoresis system (2-DE) of ovine ovarian tissue, the common protein extraction methods of animal tissue (trichloroacetic acid/acetone precipitation and direct schizolysis methods) were used to extract ovine ovarian protein, and 17-cm nonlinear immobilized PH 3-10 gradient strips were used for 2-DE. The sample handling, loading quantity of the protein sample, and isoelectric focusing (IEF) steps were manipulated and optimized in this study. The results indicate that the direct schizolysis III method, a 200-µg loading quantity of the protein sample, and IEF steps II (20°C active hydration, 14 h→500 V, 1 h→1000 V 1 h→1000-9000 V, 6 h→80,000 VH→500 V 24 h) are optimal for 2-DE analysis of ovine ovarian tissue. Therefore, ovine ovarian tissue proteomics 2-DE was preliminarily established by the optimized conditions in this study; meanwhile, the conditions identified herein could provide a reference for ovarian sample preparation and 2-DE using tissues from other animals.

Microstructural analysis of collagen and elastin fibres in the kangaroo articular cartilage reveals a structural divergence depending on its local mechanical environment.

OBJECTIVE: To assess the microstructure of the collagen and elastin fibres in articular cartilage under different natural mechanical loading conditions and determine the relationship between the microstructure of collagen and its mechanical environment. METHOD: Articular cartilage specimens were collected from the load bearing regions of the medial femoral condyle and the medial distal humerus of adult kangaroos. The microstructure of collagen and elastin fibres of these specimens was studied using laser scanning confocal microscopy (LSCM) and the orientation and texture features of the collagen were analysed using ImageJ. RESULTS: A zonal arrangement of collagen was found in kangaroo articular cartilage: the collagen fibres aligned parallel to the surface in the superficial zone and ran perpendicular in the deep zone. Compared with the distal humerus, the collagen in the femoral condyle was less isotropic and more clearly oriented, especially in the superficial and deep zones. The collagen in the femoral condyle was highly heterogeneous, less linear and more complex. Elastin fibres were found mainly in the superficial zone of the articular cartilage of both femoral condyle and distal humerus. CONCLUSIONS: The present study demonstrates that the collagen structure and texture of kangaroo articular cartilage is joint-dependent. This finding emphasizes the effects of loading on collagen development and suggests that articular cartilage with high biochemical and biomechanical qualities could be achieved by optimizing joint loading, which may benefit cartilage tissue engineering and prevention of joint injury. The existence of elastin fibres in articular cartilage could have important functional implications.

Molecular characterization and functional analysis of sheep thyroid transcription factor-1.

Thyroid transcription factor-1 (TTF-1), a member of the Nkx2 family of homeodomain-containing proteins, is involved in binding to and in activating the promoters of several important genes in the thyroid, lungs, and brain, and in regulating expression of these tissue-specific genes. We investigated potential roles of sheep (Ovis aries) TTF-1 in regulating cell fate and organ morphogenesis and in controlling puberty and reproductive capability of females. We amplified and cloned the sheep TTF-1 full-length DNA for the first time, analyzed its functional domains and regions, predicted molecular structure of its homeodomain and DNA-binding sites, and examined its expression in pituitary, brain, thyroid gland, ovary, and hypothalamus. We found that sheep TTF-1 has a high degree of homologous identity with that of other mammals, and it has several important domains including domain N, DNA-binding domain, domain C, TN-domain, domain I, and NK2-SD. The DNA-binding domain of sheep TTF-1 has 10 potential DNA-binding sites and is a novel mammalian homeodomain that shows considerable sequence homology with the corresponding rat homeodomain. Several functional regions in sheep TTF-1 share high sequence identity with rat TTF-1, indicating that these regions may have the same activity as in the rat. Expression of TTF-1 in several specific tissues implies that sheep TTF-1 in involved in sheep sexual development and reproductive capability. These results suggest a role of sheep TTF-1 in enhancing sheep reproduction performance and we propose it as a candidate gene for selection.

Growth performance, nutrient digestibility, blood parameters, and carcass characteristics by lambs fed an oregano and cobalt blend.

Shifting ruminal fermentation via feeding a blend of oregano (Organum vulgare L.) essential oils and Co-lactate (EOC; Rum-A-Fresh, Ralco, Inc. Marshall, MN) could improve lamb growth and carcass performance. Eighteen Suffolk × Little Han Tail F1 male lambs (20.3 ± 0.23 kg BW and approximately 3 months old) were randomly assigned using a completely random design to one of three treatments. Treatments were (1) EOC0: basal ration without EOC, (2) EOC4: basal ration plus 4 g/d EOC, and (3) EOC7: basal ration plus 7 g/d EOC. Initial and 24 d BW was similar (P > 0.10), but at 48 and 72 d, lambs fed EOC7 demonstrated greater (P = 0.01) BW compared with EOC0 fed lambs, while lambs fed EOC4 were intermediate and similar (P > 0.05). Average daily gains (ADGs) for 0-24 and 0-72 d were greater (P < 0.05) for lambs fed EOC4 and EOC7 compared with lambs fed EOC0, while DM intake was similar (P > 0.10). Feed conversions for 0-24 d were improved (P < 0.02) for lambs fed EOC4 and EOC7 compared with lambs fed EOC0. However, 0-72-d feed conversions were greater (P < 0.01) for lambs fed EOC7 compared to lambs fed EOC0, with lambs fed EOC4 being intermediate and similar (P > 0.05). DM, NDF, and ADF digestibilities were similar (P > 0.10) among treatments, while CP digestibility was greater (P < 0.01) for lambs fed EOC4 and EOC7 compared with lambs fed EOC0. Carcass weight and dressing percentages were improved (P < 0.01) for lambs fed EOC7 compared with lambs fed EOC0 and EOC4. Head width was greater (P > 0.01) for lambs fed EOC7 compared with lambs fed EOC0 and EOC4, while rump width was greater (P > 0.01) for lambs fed EOC4 and EOC7 compared with lambs fed EOC0. Plasma triglyceride and cholesterol concentrations were lower (P < 0.05) for lambs fed EOC4 and EOC7 compared with lambs fed EOC0, while albumin, total serum protein, and glucose concentrations were greater (P < 0.05) for lambs fed EOC4 and EOC7 compared with lambs fed EOC0. Feeding an EOC blend as an alternative antibiotic growth promoter at 4 and 7 g/d linearly improved lamb growth performance, feed conversions, frame growth, carcass weights, dressing percentages, and immunity.

LncRNA ZNFX1-AS1 targeting miR-193a-3p/SDC1 regulates cell proliferation, migration and invasion of bladder cancer cells.

OBJECTIVE: Long non-coding RNA (lncRNA) is closely associated with cancer occurrence and tumor development. However, the biological function of lncRNA ZNFX1-AS1 has not yet been reported in bladder cancer. The present study aimed to study the function of ZNFX1-AS1 in bladder cancer cells and the mechanism involved. PATIENTS AND METHODS: The expression of ZNFX1-AS1 in bladder cancer tumor tissues and cell lines was examined by qRT-PCR. The effects of ZNFX1-AS1 knockdown on cell proliferation, cell cycle, cell migration, and invasion were assessed by Cell Counting Kit-8, flow cytometry (FCM), and transwell assays. Bioinformatics analyses and Luciferase reporter assays were performed to explore the mechanism by which ZNFX1-AS1 exerted its oncogenesis role in bladder cancer. The anti-tumor effect of ZNFX1-AS1 silencing on bladder cancer in vivo was also evaluated. RESULTS: ZNFX1-AS1 was over-expressed in bladder cancer tumor tissues and cell lines. ZNFX1-AS1 expression was found to be associated with tumor size and advanced clinical stage in patients with bladder cancer. Downregulation of ZNFX1-AS1 inhibited cell proliferation, cell clone formation, migration, and invasion of bladder cancer cells. ZNFX1-AS1 was found to interact with miR-193a-3p/Syndecan 1 (SDC1). ZNFX1-AS1 expression was negatively correlated with miR-193a-3p expression, but positively correlated with SDC1 expression in bladder cancer samples. ZNFX1-AS1 knockdown also effectively suppressed tumor growth in an in vivo xenograft model. CONCLUSIONS: ZNFX1-AS1 regulated bladder cancer progression by targeting the miR-193a-3p/SDC1 axis. Our study may provide novel insights for bladder cancer prognosis and therapy.

Experimental study on early protective effect of ischemic preconditioning on rat kidney graft.

OBJECTIVE: Our aim was to investigate the mechanisms of early protection by ischemic preconditioning (IPC) of transplanted rat kidneys. MATERIALS AND METHODS: Thirty-six male donor and recipient Sprague-Dawley (SD) rats were randomly divided into 3 groups: sham-operated (A), transplantation (B), and IPC treatment (C) groups. Group A underwent exploratory laparotomy, group B received orthotopic transplantation, and group C, 15 minutes of ischemia followed by 10 minutes of reperfusion before transplantation. We measured the serum creatinine (SCr), blood urea nitrogen (BUN), and degree of kidney graft ischemic/reperfusion injury (IRI) by tumor necrosis factor-alpha (TNF-alpha) IkappaB kinase beta (IKK-beta) nuclear factor-kappa beta (NF-kappabeta) p65 subunit mRNA expressions. RESULTS: The SCr and BUN levels in groups C and B were higher than those in the sham-operated group (P < .01), without a significant difference between groups C and B at 24 hours after transplantation (P > .05). The degree of renal graft tubular injury in group C was significantly lower than that in group B (P < .01). TNF-alpha transcription levels at 24 hours after transplantation were significantly lower compared with the non-IPC group (P < .01). However, we failed to note a significant difference in IKK-beta or p65 mRNA expressions between groups C and B (P > .05). CONCLUSIONS: One cycle of preconditioning (15 min/10 min) attenuated renal graft IRI in the early phase. The inhibiting effects on TNF-alpha and the positive feedback signaling of TNF-alpha/NF-KB pathways may play important roles in renal graft protection.