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Designing an ocular drugs delivery system that can permeate the outer blood-retinal barrier (oBRB) is crucial for the microinvasive or noninvasive treatment of ocular fundus diseases. However, due to the lack of a nanocarrier that can maintain structure and composition at the oBRB, only intravitreal injection at the eyeball can deliver therapeutics directly to the ocular fundus via paracellular and intercellular routes, despite the intraocular operations risks. Here, we demonstrated tetrahedral framework nucleic acids (tFNAs) can penetrate the oBRB and deliver therapeutic nucleic acids to the retina of the rat eye in vivo following subconjunctival injection. We also discovered that tFNAs were transported via a paracellular route across the intercellular tight junctions at the oBRB. The histology analysis for ocular layers indicated that individual and aptamer/doxorubicin-loaded tFNAs penetrated all layers of the posterior segment of the eyeball to reach the innermost retina and persisted for over 3 days with minimal systemic biodistribution. We expect that the programmability and penetrability of tFNAs will provide a promising method for drug delivery across oBRB and long-term sustenance at the target site via periocular administration to various tissues.

The management of type 3 lateral condyle fractures (LCFs) remains controversial. The main goal of this study was to evaluate the feasibility of closed reduction and percutaneous pinning (CRPP) in patients with type 3 LCFs and to assess the outcome of such injuries according to the type of treatment, CRPP, or open reduction and internal fixation (ORIF).This is a retrospective review of prospectively enrolled children with type 3 LCF managed by CRPP or ORIF between 2018 and 2021. All patients were followed for at least 12 months. Patients were divided into two groups according to the type of treatment, CRPP or ORIF. Demographic characteristics were recorded for all patients. Standard radiographs were used to identify, evaluate, and classify each fracture and to detect the presence of other concomitant bone lesions. The clinical outcome was assessed according to the Hardacre et al. criteria.Seventy-eight children with type 3 LCF were included; 42 were treated by CRPP (53.8%) and 36 by ORIF (46.2%); the mean follow-up time was 17.7 months (range, 12.3-40.9). The baseline characteristics did not differ between the two groups of patients. Overall, successful CRPP could be achieved in 39 out of 42 patients (92.9%). The mean surgical time was 63.4 and 84.5 min in patients treated by CRPP and ORIF, respectively (p = 0.01). Fluoroscopy time was significantly shorter in patients managed by ORIF than in those treated by CRPP (12 versus 40 s, respectively; p 0.001). Clinical outcome according to the Hardacre et al. criteria was excellent in 37 out of 39 (94.4%) and in 35 out of 36 patients (97.2%) treated by CRPP and ORIF, respectively (p = 0.09).CRPP management of paediatric type 3 LCF has clinical and radiographic outcomes similar to ORIF; if satisfactory reduction cannot be achieved by CRPP, conversion to ORIF should be considered.

This study evaluated the outcomes of chronic Monteggia fractures (CMFs) treated by ulnar osteotomy and monolateral external fixator (MEF), and compare the outcome of gradual versus acute radial head reduction. Two groups of patients were identified. Group 1: gradual reduction of the radial head (n = 13); group 2: acute reduction (n = 6). Clinical outcome was evaluated by Kim Elbow Score, whereas radiographic outcome was assessed on plain radiographs. The effect of age, side, time from initial trauma to surgery, rate of unplanned surgery, amount of angulation and lengthening, and final outcome were evaluated. Univariate analysis was performed to identify factors associated with good radiographic outcome. Thirteen patients underwent gradual correction of the ulna. The mean duration of correction was 43.4 days (range, 21-82); the mean angulation and lengthening of the ulna were 22.8° (range, 0°-35°) and 22.2 mm (range, 12.2-40.9), respectively. Six patients underwent acute reduction intraoperatively, the mean angulation and lengthening of the ulna were 17.2° (range, 4°-33.9°) and 5.2 mm (range, 2.5-12.2), respectively. CMF treated by ulnar osteotomy and gradual distraction had better radiological outcome (Group 1; 92.3% 12/13) than those treated by acute reduction of the radial head (Group 2; 3/6, 50%) (P = 0.071). Reoperation rate was found to be significantly correlated with a fair or poor radiographic results (P = 0.016). Good clinical and radiological outcomes should be expected in CMF patients treated by gradual lengthening and angulation of the ulna with a MEF.

Tissue engineering technology provides a promising approach for large-scale bone reconstruction in cases of extensive chest wall defects. However, previous studies did not consider meticulous scaffold design specific to large-scale rib regeneration in terms of three-dimensional (3D) shape, proper porous structures, enough mechanical strength, and osteogenic microenvironments. Thus, there is an urgent need to develop an appropriate bone biomimetic scaffold (BBS) to address this problem. In this study, a BBS with controllable 3D morphology, appropriate mechanical properties, good biocompatibility and biodegradability, porous structure suitable for cell loading, and a biomimetic osteogenic inorganic salt (OIS) microenvironment was successfully prepared by integrating computer-aided design, 3D-printing, cast-molding, and freeze-drying technologies. The addition of the OIS in the scaffold substantially promoted ectopic bone regeneration in vivo, which might be attributed to the activation of osteogenic and angiogenic signaling pathways as well as upregulated expression of osteogenic genes. More importantly, dual long rib defects could be successfully repaired and medullary cavity recanalized by the rib-shaped mature cortical bone, which might be mediated by the activation of osteoclast signaling pathways. Thus, this paper presents a reliable BBS and proposes a new strategy for the repair of large-scale bone defects.

Background and Objectives: The purpose of this study was to compare the clinical and radiographic evolution of chronic Monteggia fractures (CMFs) treated by ulnar osteotomy and monolateral external fixators (MEFs) with or without angulation of the ulna during the distraction period. Materials and Methods: This retrospective study evaluated 20 children (14 boys and 6 girls) with CMFs. According to the strategy of ulnar lengthening, two groups of patients were identified: patients undergoing gradual lengthening with (Group A, n = 11) or without ulna angulation (Group B, n = 9). The mean age at the time of surgery was 7.7 years old (range, 5.4–12.9). The mean time from initial trauma to surgery was 26.3 months (range, 1–96), and the mean follow-up was 24.6 months (range, 5.5–45.4). Clinical outcomes were evaluated by Kim et al.’s Elbow Performance Score, while radiographic outcomes were assessed on plain radiographs. Results: Age at surgery, sex, laterality, time between trauma and surgery, and time of follow up in the two groups of patients showed no significant differences. The radial head was successfully reduced in 9 of 9 and 10 of 11 patients in Groups B and A, respectively (p = 1.00). The mean time to achieve radial head reduction was shorter in Group B (18.1 ± 5.3 days) than in Group A (39.2 ± 18.7 days; p = 0.004). The mean angulation of the ulna at the end of treatment was significantly lower in Group B (0.6° ± 1.1°) than in Group A (25.9° ± 6.3°; p < 0.0001). The average ulnar lengthening at the end of treatment in Group B (14.1 ± 5.8 mm) was, on average, 7.7 mm less than that in Group A (21.8 ± 9.7 mm; p = 0.05). The Kim et al. Elbow Performance Score at the last follow-up visit was comparable between the two groups of patients (p = 1.00). Conclusions: A shorter time to achieve radial head reduction and less deformity of the ulna can be expected in paediatric patients with CMFs undergoing intraoperative restoration of ulnar alignment and gradual lengthening without angulation postoperatively.

BACKGROUND: Magnetic resonance imaging (MRI) has demonstrated its potential in the evaluation of renal function. Texture analysis (TA) is a novel technique to quantify tissue heterogeneity. We aim to investigate the feasibility of using TA based on the apparent diffusion coefficient (ADC), as well as T1 and T2 maps to evaluate renal function. METHODS: Patients with impaired renal function and subjects with a normal renal function who underwent renal diffusion weighted imaging (DWI), as well as T1 and T2 mapping at 3T, were prospectively enrolled. The participants were classified into four groups according to the estimated glomerular filtration rate (eGFR, mL/min/1.73 m(2)): normal (eGFR ≥90), mildly impaired (60≤ eGFR

Cartilage defects trouble millions of patients worldwide and their repair via conventional treatment is difficult. Excitingly, tissue engineering technology provides a promising strategy for efficient cartilage regeneration with structural regeneration and functional reconstruction. Seed cells, as biological prerequisites for cartilage regeneration, determine the quality of regenerated cartilage. The proliferation, differentiation and chondrogenesis of seed cells are greatly affected by their type, origin and generation. Thus, a systematic description of the characteristics of seed cells is necessary. This article reviews in detail the cellular characteristics, research progress, clinical translation challenges and future research directions of seed cells while providing guidelines for selecting appropriate seed cells for cartilage regeneration.Cartilage defects affect millions of patients worldwide and their repair via conventional treatment is quite difficult. Excitingly, tissue engineering technology provides a promising strategy for efficient cartilage regeneration. The seed cell, as a biological prerequisite for cartilage regeneration, determines the quality of regenerated cartilage. This article reviews in detail the cellular characteristics, research progress, clinical translation challenges and future research directions of various chondrocytes, chondroprogenitor cells and stem cells. Chondrocytes, especially elastic chondrocytes, could complete subcutaneous cartilage regeneration, whereas stem cells are superior for composite defects, allografts and cartilage defects caused by inflammation. In brief, this article provides a guide for selecting appropriate seed cells for cartilage regeneration.

Some patients with developmental dysplasia of the hip (DDH) before 6 months of age successfully treated by Pavlik Harness (PH) still had persistent acetabular dysplasia (PAD). This study aimed to investigate the incidence and risk factors for PAD in patients with DDH treated by PH. We retrospectively reviewed the data of 89 patients (109 hips; mean age, 3.2 ± 1.5 months) with DDH treated by PH. Prior to treatment, all patients underwent ultrasound examination and classified according to Graf's method. PH was terminated once the hip achieved imaging recovery criteria. At final follow-up, the acetabular index (AI), center-edge angle (CEA) of Wiberg were measured on radiograph. Overall, 67/109 hips (61.5%) had successful PH treatment. Among these 67 hips, 58 hips (86.6%) achieved satisfactory outcome, nine (13.4%) had PAD. Age of the patients with PAD (4.3 ± 1 months) was significantly higher than those without PAD (2.8 ± 1.5 months) (P = 0.001). Hips with PAD had higher mean Graf grade than those with satisfactory outcome (P = 0.014). Logistic regression-confirmed age and Graf classification were risk factors for PAD. Overall, 55/67 (82.1%) met imaging recovery criteria to stop PH treatment, whereas 12/67 hips (17.9%) did not. The rate of PAD at final follow-up in patients achieving recovery criteria (4/55; 7.3%) was significantly lower than those not achieving it (41.7%) (P = 0.007). In conclusion, age and Graf classification are risk factors for PAD. If hips are not normal at the end of PH treatment, the risk of PAD increases further.

Open reduction with or without pelvic and femoral osteotomy is the accepted treatment option for children older than 18 months with developmental dislocation of the hip (DDH). However, few studies have investigated hip function in children with late DDH treated by open reduction. Therefore, we performed a retrospective study investigating the risk factors potentially associated with unsatisfactory hip function.We hypothesized that some risk factors can be associated with unsatisfactory hip function in patients with late DDH treated by open reduction.We retrospectively reviewed the clinical and radiographic data of 79 children (98 hips, mean age 39.7±18.6 months; range, 19-95.3) with late-detected DDH treated by open reduction. Acetabular index (AI), Tönnis grade, Center Edge Angle (CEA), avascular necrosis of the femoral epiphysis (AVN), and Severin radiographic grade were evaluated on radiographs. Hip function was rated according to modified Outcome Evaluation Standard for Congenital Dislocation of the Hip with a maximum score of 15. According to type of surgery, patients were divided into four groups: open reduction alone (Group A), open reduction in conjunction with pelvic osteotomy (Group B), and open reduction in conjunction with femoral osteotomy and Pemberton/Salter (Group C) or Bernese-type triple pelvic osteotomy (Group D). Analysis of variance (ANOVA), t-test, chi-square test, and multivariate regression analysis were used to evaluate the independent risk factors of unsatisfactory hip function.The mean hip function score was 10.8±2.8 (4.5-15); 61 hips (62.2%) had satisfactory function (23 [23.5%] excellent and 38 [38.8%] good), while 37 hips (37.8%) had unsatisfactory function (16 [16.3%] fair and 21 [21.4%] poor). Spearman correlation analysis indicated that age at surgery was negatively correlated with function score (r=-0.326, p=0.001). The age of patients with satisfactory hip function (34.5±14.2 months; range, 19.4-74.8) was significantly lower than those with unsatisfactory function (43.7±21 months; range, 20.6-95.3) (p=0.011). The hip function score in Group A (14.2±0.8; range, 12.8-15) was significantly higher than in the other three groups (p0.001). Hip function score in patients with type III or IV AVN was significantly lower than those without AVN, or with type II AVN (p=0.001). Multivariate regression analysis confirmed age at surgery, type of surgery and AVN were negatively correlated with hip function score. Logistic regression analysis and chi-square test confirmed age and type of surgery, and AVN were independent risk factors of unsatisfactory hip function. Multivariate regression analysis found ability to squat and amount of hip flexion and abduction, significantly decreased with age at surgery; while presence of limping, amount of hip flexion, abduction, adduction and external rotation significantly decreased with type of surgery.Age at surgery, type of surgery and severe AVN are independent risk factors for unsatisfactory hip function in patients with late-detected DDH treated by open reduction. The ability to squat, the amount of hip flexion and abduction are the main functional parameters affected by age and surgery.IV, case series.

Aims This study aimed to investigate the effect of solute carrier family 20 member 2 ( SLC20A2) gene mutation (identified from a hereditary multiple exostoses family) on chondrocyte proliferation and differentiation. Methods ATDC5 chondrocytes were cultured in insulin-transferrin-selenium medium to induce differentiation. Cells were transfected with pcDNA3.0 plasmids with either a wild-type (WT) or mutated (MUT) SLC20A2 gene. The inorganic phosphate (Pi) concentration in the medium of cells was determined. The expression of markers of chondrocyte proliferation and differentiation, the Indian hedgehog (Ihh), and parathyroid hormone-related protein (PTHrP) pathway were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Results The expression of SLC20A2 in MUT group was similar to WT group. The Pi concentration in the medium of cells in MUT group was significantly higher than WT group, which meant the SLC20A2 mutation inhibited Pi uptake in ATDC5 chondrocytes. The proliferation rate of ATDC5 chondrocytes in MUT group was greater than WT group. The expression of aggrecan (Acan), α-1 chain of type II collagen (COL2A1), and SRY-box transcription factor 9 (SOX9) were higher in MUT group than WT group. However, the expression of Runt-related transcription factor 2 (Runx2), α-1 chain of type X collagen (COL10A1), and matrix metallopeptidase 13 (MMP13) was significantly decreased in the MUT group. Similar results were obtained by Alcian blue and Alizarin red staining. The expression of Ihh and PTHrP in MUT group was higher than WT group. An inhibitor (cyclopamine) of Ihh/PTHrP signalling pathway inhibited the proliferation and restored the differentiation of chondrocytes in MUT group. Conclusion A mutation in SLC20A2 (c.C1948T) decreases Pi uptake in ATDC5 chondrocytes. SLC20A2 mutation promotes chondrocyte proliferation while inhibiting chondrocyte differentiation. The Ihh/PTHrP signalling pathway may play an important role in this process. Cite this article: Bone Joint Res 2020;9(11):751–760.

Background: To analyze the clinical data of patients aged < 6 months with developmental dislocation of the hip (DDH) treated with Pavlik Harness (PH) in order to identify the best time to terminate PH treatment. Method: Fifty-four patients (47 females, 7 males; 63 hips) met the inclusion criteria and were included in the study; there were 33 (61.1%) left, 12 (22.2%) right and 9 (16.7%) bilateral DDH. The mean age at diagnosis was 11.8 ± 5.9 weeks (range, 1.4–25.5). All patients underwent fulltime PH treatment for about three months. At completion of PH treatment, patients were then divided into Group A, including patients with clinically stable hip joint and Graf type-I hip on ultrasound (US), and Group B, including patients with clinically stable hip joint and well-reduced hip on anterior-posterior (AP) radiographs without acetabular dysplasia. Six months after completion of PH treatment, the presence/absence of residual acetabular dysplasia (RAD) was evaluated on AP pelvis radiographs. The t-test and chi-square test were used to compare the differences in age, gender, side, Graf classification and RAD rate between the two groups of patients. Results: At completion of PH treatment, 45 hips were in Group A and 18 in Group B. There were no significant differences in age, gender, side, preoperative alpha angle and Graf classification between the two groups. Six months after discontinuation of PH, the AI in Group A (27.1° ± 6.8°) was significantly higher than that in Group B (21.9° ± 3.5°; p = 0.001); moreover 23 hips (51.1%) in Group A developed RAD compared to one hip in Group B (5.6%; p = 0.001). Among Group A patients, those with RAD were significantly older (13.7 ± 4.9 weeks) than those with normal hips (7.6 ± 3.8 weeks; p < 0.001); the incidence of RAD was significantly lower in patients with Graf type-II D hips (22.2%) than in patients with Graf type-III (70%) and type-IV hips (71.4%; p = 0.006). However, logistic regression analysis identified age as the only risk factor for RAD. All 24 hips with RAD (24/63, 38.1%) were treated with abduction braces. At final follow-up, AI in Group A (20.5° ± 3.3°) was not significantly different from that in Group B (21.9° ± 3.3°; p = 0.132). At the last follow-up visit, five hips (11.1%) in Group A still had RAD, compared to none in Group B (p = 0.31). Conclusions: In patients with DDH treated by PH, Graf type-I on US is not an absolute timing to terminate PH treatment. In addition, patients ≥ 13 weeks had a high risk of RAD despite PH treatment as 51.1% of infants developed RAD during follow up. Follow-up radiographs should be requested in all patients achieving Graf type-I hips at completion of PH treatment.

This study aims to evaluate (1) the probability to achieve normal pelvic radiographs in children with developmental dysplasia of the hip (DDH) treated by closed reduction and (2) the amount of time needed to achieve normal pelvic radiographs and to assess what factors influence both probability and time to achieve normal radiographic parameters following CR and spica cast immobilization for DDH. We retrospectively reviewed 436 patients (393 girls, 43 boys; 507 hips) with DDH treated by closed reduction (CR). Tonnis grade, AVN, acetabular index (AI), centre-edge angle (CEA), and Severin radiographic grade were evaluated on plain radiographs. Criteria to rate pelvis radiographs as normal were established. Cox regression was used to evaluate the factors influencing the probability and the time to achieve normal radiographs. According to our criteria, 167 hips (32.9%) achieved normal radiographic parameters during follow-up. The overall amount of time to achieve normal pelvis radiographs was 36.1 ± 15.5 months. Patients older than 24 months of age at the time of CR needed longer time to achieve normal radiographic parameters (55.2 ± 28 months) compared with other age groups. Cox regression analysis suggested the overall cumulative probability of recovery increased by 46% at five years following CR, then it tended to plateau with an annual increase less than 5%. Age older than 24 months, bilateral dislocation, pre-operative AI greater than 40°, and AVN were risk factors for reduced probability of achieving normal radiographic parameters. The cumulative probability of achieving normal pelvis radiographs increases linearly during the first five years following CR, then it tends to plateau. Age older than 24 months and Tonnis grade III and IV are associated with longer time to achieve normal radiographic parameters. Age older than 24 months, bilateral dislocation, pre-operative AI greater than 40°, and AVN are risk factors for reduced probability of achieving normal radiographic parameters in children with DDH treated by closed means.

Heat stress (HS) and secondary restricted blood flow to the intestines cause dysfunction of the intestinal epithelial barrier. Tight junctions (TJs) are essential to maintain intestinal integrity. l-Arginine has beneficial effects on gut functions. However, the underlying mechanisms remain largely unknown. This study tested the hypothesis that l-arginine regulates the TJ network by activating AMP-activated protein kinase (AMPK) signaling, which in turn improves intestinal barrier functions under HS. IEC-6 cells and rat small intestines were used as experiment models of heat stress. AICAR and dorsomorphin were used to activate and inhibit the AMPK pathway, respectively. Cell proliferation, apoptosis, differential gene expression and KEGG pathway analysis, intestinal paracellular permeability, intestinal morphology, and expression of HSP and TJ proteins, and p-AMPK were determined. l-Arginine promoted cell proliferation and reduced apoptosis after heat exposure at an optimal concentration of 5 mmol. Transcriptome sequencing analysis revealed that differentially expressed genes associated with the HSP family and TJs were elevated by l-arginine. According to KEGG pathway analysis, l-arginine activated the AMPK signaling pathway. In vivo, intestinal damage resulted in obvious morphological changes as well as apoptosis with TUNEL and caspase-3 staining under HS and dorsomorphin treatments. Furthermore, HS and dorsomorphin increased the serum D-lactate concentration, diamine oxidase activity, and mRNA expression level of MLCK (P l-arginine and AICAR treatments reduced intestinal injury, maintained intestinal permeability, and increased the villus/crypt ratio under hyperthermia. l-Arginine had the same effect as AICAR both in vitro and in vivo, namely increasing p-AMPK protein expression. l-Arginine and AICAR also upregulated the mRNA expression level of HSP70 and HSP90, and downregulated mRNA expression of MLCK (P l-arginine and AICAR. Our findings indicate that activation of AMPK signaling by l-arginine is associated with improved intestinal mucosal barrier functions by enhancing the expression of TJs in rat small intestines and IEC-6 cells during HS.

BACKGROUND To compare the effects of two types of cast immobilization (human position cast and dynamic cast) on hip development in children with Developmental dysplasia of the hip (DDH) after closed reduction (CR). METHODS A retrospective study of 60 children (64 hips) with DDH who underwent CR and cast immobilization between January 2015 and December 2016 at our Institution was performed. The average age at the time of CR was 14.6 months (range, 6.1-23.5). Fifty-seven females and 3 males were included. According to the technique of cast immobilization, two groups of patients could be identified: patients with DDH managed by human position cast immobilization (Group A: 32 patients, 34 hips) and patients with DDH treated by dynamic cast immobilization (Group B: 28 patients, 30 hips). Hip joint distance (HJD) after CR was measured on MRI. Acetabular index (AI) and Acetabular Depth Radio (ADR) were measured of anterior-posterior (AP) radiographs before and 3 months after CR; AI and central edge angle (CEA) were measured last follow-up AP radiographs. The presence of subluxation or dislocation and avascular necrosis (AVN) at the last follow-up visit was also evaluated. RESULTS The patients were comparable regarding to sex, side, age, Tonnis degree, AI, and ADR before the reduction between two groups. There was no significant difference in HJD improvement between the two groups 6 weeks following closed reduction. The AI(27.5±5.1°) of group B was significantly lower than those of Group A (31±4.9°) (p=0.03) when cast was removed 3 months after CR. At the last follow-up, the incidence of AVN was similar between the two groups of patients (Group A: 11.8% versus Group B: 13.3%), and the incidence of subluxation or dislocation (Group A: 8.8% versus Group B: 10%). At last follow-up visit, the AI (23.7±5.4°) in Group B was significantly lower than in Group A (26.9±4.1°) (p=0.02). CONCLUSIONS Dynamic cast immobilization promotes acetabular development following CR in patients aged 6 to 24 months with DDH. Dynamic cast immobilization does not increase the risk of dislocation or subluxation, nor of AVN.

Neurofibromatosis type I (NF1) is an autosomal dominant genetic disease that is caused by mutations in the NF1 gene. Various studies have previously demonstrated that the mTOR complex 1 signaling pathway is essential for the NF1-modulated osteogenic differentiation of bone mesenchymal stem cells (BMSCs). Additionally, the mTOR signaling pathway plays a notable role in autophagy. The present study hypothesized that NF1 could modulate the osteogenic differentiation of BMSCs by regulating the autophagic activities of BMSCs. In the present study, human BMSCs were cultured in an osteogenic induction medium. The expression of the NF1 gene was either knocked down or overexpressed by transfection with a specific small interfering RNA (siRNA) targeting NF1 or the pcDNA3.0 NF1-overexpression plasmid, respectively. Autophagic activities of BMSCs (Beclin-1, P62, LC3B I, and LC3B II) were determined using western blotting, electron microscopy, acridine orange (AO) staining and autophagic flux/lysosomal detection by fluorescence microscopy. In addition, the autophagy activator rapamycin (RAPA) and inhibitor 3-methyladenine (3-MA) were used to investigate the effects of autophagy on NF1-modulated osteogenic differentiation in BMSCs. Inhibiting NF1 with siRNA significantly decreased the expression levels of autophagy markers Beclin-1 and LC3B-II, in addition to osteogenic differentiation markers osterix, runt-related transcription factor 2 and alkaline phosphatase. By contrast, overexpressing NF1 with pcDNA3.0 significantly increased their levels. Transmission electron microscopy, AO staining and autophagic flux/lysosomal detection assays revealed that the extent of autophagosome formation was significantly decreased in the NF1-siRNA group but significantly increased in the NF1-pcDNA3.0 group when compared with the NC-siRNA and pcDNA3.0 groups, respectively. In addition, the activity of the PI3K/AKT/mTOR pathway [phosphorylated (p)-PI3K, p-AKT, p-mTOR and p-p70S6 kinase] was significantly upregulated in the NF1-siRNA group compared with the NC-siRNA group, and significantly inhibited in the NF1-pcDNA3.0 group, compared with the pcDNA3.0 group. The knockdown effects of NF1-siRNA on the autophagy and osteogenic differentiation of BMSCs were reversed by the autophagy activator RAPA, while the overexpression effects of NF1-pcDNA3.0 on the autophagy and osteogenic differentiation of BMSCs were reversed by the autophagy inhibitor 3-MA. In conclusion, results from the present study suggest at the involvement of autophagy in the NF1-modulated osteogenic differentiation of BMSCs. Furthermore, NF1 may partially regulate the autophagic activity of BMSCs through the PI3K/AKT/mTOR signaling pathway.

In order to evaluate the pathogenicity of Senecavirus A (SVA) to weaned piglets preliminarily, 28-day-old weaned piglets were challenged with SVA by intramuscular injection. The clinical manifestations, antibody levels, and tissue viral load of infected piglets were detected. The results indicated that the piglets challenged with SVA CH/FuJ/2017 showed drowsiness, lameness, oral blisters, diarrhea, and other clinical signs. Lesions on the hooves were observed. Red spots or plaques were initially observed on the hoof and then developed into blisters that cracked and gradually formed scab. The symptoms and signs were relieved after 8 days post-infection (dpi). The sentinel piglet, feeding together with the challenged piglets, showed similar clinical signs with the challenged piglets after 3 dpi. Monitoring of antibody levels showed that anti-SVA antibody could be detected at 5 dpi by competition enzyme-linked immunosorbent assay (cELISA) method, and neutralizing antibody could be detected after 7 dpi. Analysis of viral tissue distribution and viral load indicated that SVA could replicate in the liver, spleen, lung, kidney, and lymph node. In all, Senecavirus disease was successfully replicated by SVA CH/FuJ/2017 isolate, which verified the clinical manifestations of SVA infection in weaned piglets, and provided a foundation for further SVA pathogenesis and vaccine development.

Trunk movements during quiet and deep breathing in untreated and in operated patients with scoliosis are not well defined. To evaluate sagittal and transverse plane cross-sectional variations of the trunk during quiet and deep breathing by optical reflective motion analysis (ORMA) in children with scoliosis. Twenty-one patients were divided into three groups: normal subjects (A; n = 6), subjects with untreated scoliosis50° (B; n = 7) and operated patients (C; n = 8). Standing and sitting height, T1-T12 and L1-L5 length, arm span, chest perimeter, weight and BMI were recorded. Trunk movements of all patients, during quiet and deep breathing, were measured with a 10-camera 3D ORMA system (82 markers) with the subjects in a standard standing position. Groups were comparable with respect to age, sex, height, arm span and weight (P 0.05). Significant differences were found in Cobb angle, chest perimeter and BMI (P 0.05). Trunk sagittal and transverse plane movements during quiet and deep breathing decreased significantly in group B and group C when compared to group A (P 0.05). Surgery does not completely eliminate sagittal and transverse plane kinematics, although the spine is rendered more rigid. This preliminary study involving a relatively limited number of patients outlines ORMA is a useful tool for analyzing sagittal and transverse plane motion abnormalities of the trunk. Trunk kinematics was altered during breathing in unoperated patients with scoliosis50°. Operated subjects had sagittal plane trunk kinematics closer to normal subjects, although changes were less pronounced at the level of the convex side.

Aims This study aimed to investigate the effect of solute carrier family 20 member 2 (SLC20A2) gene mutation (identified from a hereditary multiple exostoses family) on chondrocyte proliferation and differentiation. Methods ATDC5 chondrocytes were cultured in insulin-transferrin-selenium medium to induce differentiation. Cells were transfected with pcDNA3.0 plasmids with either a wild-type (WT) or mutated (MUT) SLC20A2 gene. The inorganic phosphate (Pi) concentration in the medium of cells was determined. The expression of markers of chondrocyte proliferation and differentiation, the Indian hedgehog (Ihh), and parathyroid hormone-related protein (PTHrP) pathway were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Results The expression of SLC20A2 in MUT group was similar to WT group. The Pi concentration in the medium of cells in MUT group was significantly higher than WT group, which meant the SLC20A2 mutation inhibited Pi uptake in ATDC5 chondrocytes. The proliferation rate of ATDC5 chondrocytes in MUT group was greater than WT group. The expression of aggrecan (Acan), α-1 chain of type II collagen (COL2A1), and SRY-box transcription factor 9 (SOX9) were higher in MUT group than WT group. However, the expression of Runt-related transcription factor 2 (Runx2), α-1 chain of type X collagen (COL10A1), and matrix metallopeptidase 13 (MMP13) was significantly decreased in the MUT group. Similar results were obtained by Alcian blue and Alizarin red staining. The expression of Ihh and PTHrP in MUT group was higher than WT group. An inhibitor (cyclopamine) of Ihh/PTHrP signalling pathway inhibited the proliferation and restored the differentiation of chondrocytes in MUT group. Conclusion A mutation in SLC20A2 (c.C1948T) decreases Pi uptake in ATDC5 chondrocytes. SLC20A2 mutation promotes chondrocyte proliferation while inhibiting chondrocyte differentiation. The Ihh/PTHrP signalling pathway may play an important role in this process. Cite this article: Bone Joint Res 2020;9(11):751–760.

Background: Although the main causative genes for hereditary multiple exostoses (HME) are EXT-1 and EXT-2, there are still many HME patients without EXT-1 and EXT-2 mutations. This study aimed to identify novel candidate genes for the development of HME in patients without EXT-1 and EXT-2 mutations.Methods: Whole-exome sequencing was performed in a typical Chinese HME family without EXT-1 and EXT-2 mutations, followed by a combined bioinformatics pipeline including annotation and filtering process to identify candidate variants. Candidate variants were then validated by Sanger sequencing.Results: A total of 1830 original variants were revealed to be heterozygous mutations in the three patients suffering from HME that were not present in the healthy controls. Two mutations (c.C1849T in SLC20A2 and c.G506A in LETM1) were identified as a possible causative variant for HME through a bioinformatics filtering procedure and harmful prediction. Sanger sequencing results confirmed these two mutations in all patients with HME. A mutation in SLC20A2 (c.C1849T) led to a change in amino acid (p.R617C), which may be involved in the development of HME by inducing metabolic disorders of phosphate and abnormal proliferation and differentiation in chondrocytes.Conclusions: The present study revealed two mutations [SLC20A2 (c.C1849T) and LETM1 (c.G506A) in a Chinese HME family. The mutation in SLC20A2 (c.C1849T) is more likely to be involved in the development of HME.

The aim of this study was to investigate the effects of Clostridium butyricum (C. butyricum) on the performance, serum lipid metabolism, muscle morphology, meat quality, and fatty acid profiles of Peking ducks. A total of 1,500 Peking ducks were randomly divided into five groups with five replicates and were fed a non-antibiotic basal diet (Control) or a basal diet supplemented with either 200, 400, or 600 mg/kg of C. butyricum (2.0 × 109 CFU/g) or 150 mg of aureomycin/kg for 42 d. Compared with the control group, supplementation with C. butyricum increased the average daily weight gain but reduced the feed/gain ratio from 1 to 42 d of age. Similarly, dietary C. butyricum increased the activities of antioxidant enzymes but decreased the malondialdehyde (MDA) and lipid metabolites concentration. C. butyricum supplementation increased the muscle pH value at 45 min postmortem, the redness of the meat, and the contents of inosine acid (IMP) and intramuscular fat (IMF) in Peking ducks. By contrast, C. butyricum supplementation lowered the lightness, drip loss, and the shear force of breast meat. Supplementation with C. butyricum increased the concentrations of essential amino acids and flavor amino acids, as well as arachidonic acid (AA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and total polyunsaturated fatty acids (PUFA) in breast muscle. Dietary C. butyricum could positively improve performance, lipid metabolism, meat quality, and the amino acid and fatty acid composition in a dose-dependent manner. Therefore, C. butyricum is proposed as a feasible alternative feed additive for the production of healthier Peking duck meat with favorable properties.

Increased femoral anteversion can be associated with hip instability, redislocation after closed reduction, and subsequent early degenerative arthritis. Our study compared proximal femoral anteversion of affected and unaffected sides of patients with unilateral developmental dysplasia of the hip (DDH) on two-dimensional computed tomography. The primary aim was to evaluate whether femoral anteversion at the time of treatment affected the outcome of patients with unilateral DDH treated by closed reduction.A retrospective review of 89 patients (82 females; 53 left; mean age: 26.6 months) with unilateral DDH was performed. Anteversion angle (AA) of the femur and acetabular index (AI) of both affected (AAa; AIa) and unaffected (AAu; AIu) hips were measured on two-dimensional CT scan performed no more than seven days prior to the index surgical procedure.Among the 89 patients, 50 underwent closed reduction (56.2%), 38 underwent open reduction with or without pelvic osteotomy (42.7%), and one patient refused treatment (1.1%). Overall, the mean AAa was 28.1° ± 10.2° (range: 6.3°-54°) and mean AAu was 25.2° ± 9.9° (range: 1.9°-52.5°) (t = 3.2, p = 0.002). Tönnis type 2 hips did not show any statistically significant difference between AAa and AAu (p = 0.386), while Tönnis types 3 and 4 hips had significantly higher AAa than did AAu (t = 3.7, p = 0.001). There were significant correlations between age and AAa (coefficient = 0.4; p 0.001) and AAu (coefficient = 0.304; p = 0.004). Correlation analysis showed that AIa did not improve with age in any Tönnis group (r: - 0.24, p = 0.823; F = 0.039, p = 0.962). AAa, AIa, AAD, AID, and Tönnis grade distribution were similar in patients with good (no redislocation) and poor outcomes (redislocation) (p 0.05).In patients with unilateral DDH, anteversion angle (AA) was found to be significantly different between affected and unaffected sides. However, the difference had very limited or no clinical significance, as redislocation/sub-luxation was not influenced by AA values.

Background Bovine viral diarrhoea virus (BVDV) is the member of the genus Pestivirus within the Flaviviridae family and responsible for severe economic losses in the cattle industry. BVDV can employ ‘infect-and-persist’ strategy and ‘hit-and-run’ strategy to remain associated with hosts and thus contributes to BVDV circulation in cattle herds. BVDV have also evolved various strategies to evade the innate immunity of host. To further understand the mechanisms by which BVDV overcomes the host cell innate immune response and provide more clues for further understanding the BVDV-host interaction, in this descriptive study, we conducted a investigation of differentially expressed genes (DEGs) of the host during BVDV infection by RNA-Seq analysis. Results Our analysis identified 1297, 1732, 3072, and 1877 DEGs in the comparison groups mock vs. MDBK cells infected with BVDV post 2 h (MBV2h), mock vs. MBV6h, mock vs. MBV12h, and mock vs. MBV24h, respectively. The reproducibility and repeatability of the results were validated by RT-qPCR. Enrichment analyses of GO annotations and KEGG pathways revealed the host DEGs that are potentially induced by BVDV infection and may participate in BVDV-host interactions. Protein-protein interaction (PPI) network analyses identified the potential interactions among the DEGs. Our findings suggested that BVDV infection induced the upregulation of genes involved in lipid metabolism. The expression of genes that have antiviral roles, including ISG15, Mx1, OSA1Y, were found to be downregulated and are thus potentially associated with the inhibition of host innate immune system during BVDV infection. The expression levels of F3, C1R, KNG1, CLU, C3, FB, SERPINA5, SERPINE1, C1S, F2RL2, and C2, which belong to the complement and coagulation signalling cascades, were downregulated during BVDV infection, which suggested that the complement system might play a crucial role during BVDV infection. Conclusion In this descriptive study, our findings revealed the changes in the host transcriptome expression profile during BVDV infection and suggested that BVDV-infection induced altering the host’s metabolic network, the inhibition of the expression of antiviral proteins and genes within the complement system might be contributed to BVDV proliferation. The above findings provided unique insights for further studies on the mechanisms underlying BVDV-host interactions.

Background Open reduction with or without pelvic and femoral osteotomy is the accepted treatment option for children older than 18 months with developmental dislocation of the hip (DDH). However, few studies have investigated hip function in children with late DDH treated by open reduction. Therefore, we performed a retrospective study investigating the risk factors potentially associated with unsatisfactory hip function. Hypothesis We hypothesized that some risk factors can be associated with unsatisfactory hip function in patients with late DDH treated by open reduction. Patients and methods We retrospectively reviewed the clinical and radiographic data of 79 children (98 hips, mean age 39.7 ± 18.6 months; range, 19–95.3) with late-detected DDH treated by open reduction. Acetabular index (AI), Tonnis grade, Center Edge Angle (CEA), avascular necrosis of the femoral epiphysis (AVN), and Severin radiographic grade were evaluated on radiographs. Hip function was rated according to modified Outcome Evaluation Standard for Congenital Dislocation of the Hip with a maximum score of 15. According to type of surgery, patients were divided into four groups: open reduction alone (Group A), open reduction in conjunction with pelvic osteotomy (Group B), and open reduction in conjunction with femoral osteotomy and Pemberton/Salter (Group C) or Bernese-type triple pelvic osteotomy (Group D). Analysis of variance (ANOVA), t-test, chi-square test, and multivariate regression analysis were used to evaluate the independent risk factors of unsatisfactory hip function. Results The mean hip function score was 10.8 ± 2.8 (4.5–15); 61 hips (62.2%) had satisfactory function (23 [23.5%] excellent and 38 [38.8%] good), while 37 hips (37.8%) had unsatisfactory function (16 [16.3%] fair and 21 [21.4%] poor). Spearman correlation analysis indicated that age at surgery was negatively correlated with function score (r = −0.326, p = 0.001). The age of patients with satisfactory hip function (34.5 ± 14.2 months; range, 19.4–74.8) was significantly lower than those with unsatisfactory function (43.7 ± 21 months; range, 20.6–95.3) (p = 0.011). The hip function score in Group A (14.2 ± 0.8; range, 12.8–15) was significantly higher than in the other three groups (p Discussion Age at surgery, type of surgery and severe AVN are independent risk factors for unsatisfactory hip function in patients with late-detected DDH treated by open reduction. The ability to squat, the amount of hip flexion and abduction are the main functional parameters affected by age and surgery. Level of evidence IV, case series.

This study aimed to investigate the effects of chromic chloride (CrCl3) on Ca, Mg, Mn, Fe, Cu, and Zn contents in the brain and serum of chicken. Seventy-two chickens were randomly divided into four groups and treated with different doses of CrCl3 via drinking water: 0, 1/8, 1/4, and 1/2 LD50 for 42 days. The contents of the elements were evaluated through inductively coupled plasma mass spectrometry. Results showed that Cr contents in the brain and serum were higher than those in the control groups, although no significant dose-dependent changes (P > 0.05) in brain of the Cr-treated groups were observed at 42 days. As exposure time was prolonged and CrCl3 dosage was increased, Ca contents increased (P

This study aimed to investigate the effect of age on the rates of redislocation, avascular necrosis (AVN) of the femoral head, and final radiographic outcomes in patients with developmental dysplasia of the hip (DDH) treated with closed reduction. A total of 308 hips (276 patients) with DDH treated with closed reduction were included and divided into three groups according to age (

The aim of this study was to compare the clinical and radiological outcomes in children with acute Monteggia fracture treated by two different surgical techniques. Twenty-six children treated by either closed reduction and external fixation (group A) or elastic stable intramedullary nails (group B) were retrospectively reviewed. Clinical outcome was evaluated with the Quick DASH questionnaire. Overall, both surgical techniques provided similar excellent healing rates and good clinical outcomes (P > 0.05), as well as radiological outcomes (P > 0.05). Radiographically, no secondary displacement, wire migration, consolidation delays, non-union, malunion or re-fractures were noted. However, two patients (7.6%), one in group A (1/15; 6.7%) and one in group B (1/11; 9.1%), developed heterotopic ossification at the level of the ulna. Clinical functional outcomes were similar between the two groups of patients. The mean overall Quick DASH score was 0.79 (range, 0-18.2). However, six in group A (40%) and one in group B (9.1%) complained of the clinical appearance and had intermittent residual pain on the injured side. In conclusion, both elastic stable intramedullary nailing and external fixation can achieve satisfactory clinical and radiographic outcomes in children with acute Monteggia fractures. However, the rate of residual pain and less satisfactory clinical appearance was higher in patients managed by external fixation compared to those treated by elastic stable intramedullary nailing. According to our results, elastic stable intramedullary nailing should be used as primary treatment option in children with acute Monteggia fracture requiring surgical management.

Background Congenital talipes equinovarus (CTEV) is the most common congenital deformity in children, and muscular dysplasia plays a potential role in the etiology of CTEV. Notably, previous studies have found that HOXA9 rs3801776 and TPM2 rs2025126 genetic polymorphisms play important roles in regulating muscle development in Caucasian children; however, there is a lack of investigations conducted in Chinese children. Methods We conducted a hospital-based, case-control study of 189 children with CTEV and 457 CTEV-free children aiming to examine the associations between these two polymorphisms and CTEV susceptibility. The rs3801776 (G>A) and rs2025126 (G>A) polymorphisms were genotyped using TaqMan. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to measure the associations between the selected polymorphisms and CTEV susceptibility. Results We found that rs3801776A was associated with increased CTEV risk (GA versus GG: adjusted OR = 1.81, 95% CI = 1.22-2.69, p = 0.0031; AA versus GG: adjusted OR = 2.19, 95% CI = 1.28-3.73, p = 0.0041; GA/AA versus GG: adjusted OR = 1.89, 95% CI = 1.29-2.76, p = 0.0010). In a stratified analysis, the risk effect of rs3801776 GA/AA was observed in both unilateral and bilateral patients. Conclusions The present study suggests that the rs3801776 G>A polymorphism is associated with CTEV risk in Chinese children; however, this conclusion should be validated in larger studies.

Heat stress (HS) and the associated restricted blood flow to the intestine have been proven to destroy intestinal integrity. Considering the beneficial properties of L-arginine on gut function, we investigated the protective effects of L-arginine on the intestine under HS conditions. In vivo, the serum cortisol level and the rectal temperature increased in response to HS. Under HS, the intestinal damage showed obvious morphological changes. Furthermore, HS decreased the mRNA and protein expression levels of Nurr1, ZO-1, occludin, claudin-6 and E-cadherin, increased the mRNA expression of NF-κB and IL-1β, and increased the protein expression of cleaved caspase-3. In contrast, L-arginine supplementation maintained intestinal integrity and increased the villus/crypt ratio. L-arginine also suppressed the expression of inflammation-related genes and the protein expression of cleaved caspase-3, whereas it upregulated the mRNA and protein expression of tight junction proteins and LC3B protein expression. In vitro, L-arginine attenuated HS-induced apoptosis as demonstrated by flow cytometry and decreased cleaved caspase-3 protein expression. L-arginine induced autophagy, which was demonstrated by decreased expression of p62 and p-mTOR/mTOR, and increased expression of LC3B. The protein expression levels of TJ proteins also enhanced by L-arginine in IEC-6 cells. Taken together, these results suggest that L-arginine can alleviate intestinal damage and protect the intestinal integrity by suppressing local inflammation response, promoting the production of TJs and facilitating autophagy under HS conditions.

Additional file 2: Figure S1. Venn analysis of functional annotation and classification within GO, COG and KEGG database. Figure S2. Functional annotation of the genes obtained from RNA-Seq. The different colours of the pie chart represent different GO terms, and the area represents the relative proportion of genes/transcripts in the GO Term. The number of genes were shown behind the subsets. Go terms of level 2 were annotated within the three Go categories, biological progress (BP), cellular progress (CC), and molecular function (MF). The functional annotation was performed using Blast2go. Figure S3. Function classification in Clusters of Orthologous Groups of Proteins (COG) of genes from RNA-Seq. Capital letters on x axis indicated the COG categories as listed on the right of histogram; y axis indicated the number of gene. Figure S4. Pathway annotation of genes from RNA-Seq. KEGG Pathway of level 2 within the seven categories of KEGG pathway as listed on the right of histogram were annotated. x axis indicated the number of genes annotated to the pathway; y axis indicated the name of the KEGG pathway. Figure S5. Venn analysis of differential expression genes from group Mock vs. MBV2h, Mock vs. MBV6h, Mock vs. MBV12h, Mock vs. MBV24h. Figure S6. Dynamic changes of BVDV RNA in MDBK cells infected with 10 MOI BVDV. a: BVDV positive strand RNA; b: BVDV negative strand RNA. Copies of BVDV positive or negative strand RNA was quantified by strand specific SYBR Green real-time PCR and shown as copies of positive or negative strand RNA of BVDV per microgram total RNA of cells. Figure S7. GO enrichment analysis of DEGs in comparison groups MBV2h vs. MBV6h(a), MBV6h vs. MBV 12 h(b), MBV12h vs. MBV24h(c). GO terms are on the x axis. Enrichment ratio of genes shown as GO terms for BP, CC, and MF. * means GO terms with significant enrichment. Figure S8. KEGG Pathway enrichment analysis of DEGs in comparison groups MBV2h vs. MBV6h (a), MBV6h vs. MBV12h (b), MBV12h vs. MBV24h (c). The name of KEGG pathway are on the x axis. Enrichment ratio of genes shown as the name of KEGG pathway for seven categories. * means KEGG Pathway with significant enrichment. Figure S9. Protein–Protein Interaction Network Analysis of DEGs with FDR ≤ 0.05 and |Log2FC| ≥ 2 in comparison group Mock vs. MBV2h. OAS1Y, FOS and EGR1 were three genes with the most node degree in the analysis, labelled in red blot, green blot and yellow blot, respectively. Figure S10. Protein–Protein Interaction Network Analysis of DEGs with FDR ≤ 0.05 and |Log2FC| ≥ 2 in comparison group Mock vs. MBV24h. OAS1Y and FOSB were the two genes with the most node degree, labeled in red blot and yellow blot, respectively.

To obtain an effective vaccine candidate against duck Tembusu viral (DTMUV) disease which causes egg-drop and great economical loss in the Chinese duck industry, liposome vaccines containing recombinant E protein were prepared and assessed in this study. The recombinant plasmid (PET28a-E) was constructed and transformed into BL21 (DE3) cells to produce E proteins. The recombinant E proteins were purified and entrapped by liposomes through reverse-phase evaporation. Eighty-four cherry valley ducks were randomly divided into seven groups and inoculated intramuscularly at one- or seven-day-old with liposomes-E protein or Freund's adjuvant-E protein vaccine. Blood samples were collected from the first week to the tenth week for serum antibody, plasma for viremia, as well as oropharyngeal and cloacal swabs for virus shedding analyses after being challenged with a 10(2.4) 50% tissue culture infective dose (TCID50) of duck Tembusu virus. Results showed that serum antibody level of the liposomes vaccine was higher than the Freund's adjuvant vaccine, and inoculating twice was superior to once; furthermore, the viremia and virus shedding tests also proved that the liposomes vaccine can provide complete protection against DTMUV challenge. These results demonstrated that the liposomes-E protein vaccine could be used as a potential candidate vaccine to prevent DTMUV infection in ducks.

This experiment was conducted to investigate the oxidative stress in chickens exposed to different concentrations of chromium trichloride (CrCl3) in drinking water. Seventy-two Hylan Brown male chickens were randomly divided into four groups: three experimental groups and one control group. The experimental groups were exposed to three different doses (50 % LD50, 25 % LD50, and 12.5 % LD50) of CrCl3 mg/kg body weight for 42 days, while the control group was given the equivalent water. The activities of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) and non-enzymatic index (glutathione, total antioxidant capacity, malondialdehyde, and hydrogen peroxide) were measured after obtaining the brain samples. Results suggested that 50 % LD50 chromium(III) significantly increased (P

This study aimed to investigate the effects of oral administration of trivalent chromium on the contents of Ca, Mg, Mn, Fe, Cu, and Zn in the heart, liver, and kidney. Different levels of 1/8, 1/4, and 1/2 LD50 (LD50 = 5000 mg/kg body mass) CrCl3 milligrams per kilogram body mass daily were added into the water to establish the chronic poisoning model. Ca, Mg, Mn, Fe, Cu, and Zn were detected with the flame atomic absorption spectrometry in the organs exposed 14, 28, and 42 days to CrCl3, respectively. Results showed that Cr was accumulated in the heart, liver, and kidney significantly (P

Background Increased attention is being paid to breast muscle yield and meat quality in the duck breeding industry. Our previous report has demonstrated that dietary Clostridium butyricum (C. butyricum) can improve meat quality of Pekin ducks. However, the potential biological processes and molecular mechanisms that are modulated by dietary C. butyricum in the breast muscle of Pekin ducks remain unknown. Results Supplementation with C. butyricum increased growth performance and meat yield. Therefore, we utilized de novo assembly methods to analyze the RNA-Seq transcriptome profiles in breast muscle to explore the differentially expressed genes between C. butyricum-treated and control Pekin ducks. A total of 1119 differentially expressed candidate genes were found of which 403 genes were significantly up-regulated and 716 genes were significantly down-regulated significantly. qRT-PCR analysis was used to confirm the accuracy of the of RNA-Seq results. GO annotations revealed potential genes, processes and pathways that may participate in meat quality and muscle development. KEGG pathway analysis showed that the differentially expressed genes participated in numerous pathways related to muscle development, including ECM-receptor interaction, the MAPK signaling pathway and the TNF signaling pathway. Conclusions This study suggests that long-time dietary supplementation with C. butyricum can modulate muscle development and meat quality via altering the expression patterns of genes involved in crucial metabolic pathways. The findings presented here provide unique insights into the molecular mechanisms of muscle development in Pekin ducks in response to dietary C. butyricum. Electronic supplementary material The online version of this article (10.1186/s12864-018-5261-1) contains supplementary material, which is available to authorized users.

Table S1. Composition and nutrient level of diets (air-dry basis). Table S2. Quality control of each RNA sample for sequencing. Table S3. Evaluation of clean data of each sample in this study. Table S4. Effects of Clostridium butyricum on breast meat quality of Pekin ducks. Table S5. Mapping ratios of each sample to the reference duck genome in this study. Table S6. Mapping Statics and ratios of each sample to de novo assembled transcripts in this study. (DOCX 29 kb)

Figure S1. Species distribution of the top BLAST hits. Figure S2. Different levels of GO term function classification. Figure S3. The top 20 pathways according to numbers of unigene annotation. Figure S4. The numbers of up- or down-regulated genes annotated to each GO term function. (DOCX 1113 kb)

This study aims to investigate the oxidative stress and hepatocellular injury induced by Cr(3+) in chicken. Different doses of CrCl3 solutions (50% LD50 , 25% LD50 , and 12.5% LD50) and equivalent water were orally administered to chicken. Chicken liver samples were measured for the activities of antioxidant enzymes, the contents of glutathione, total antioxidant capacity (T-AOC), malondialdehyde (MDA), and hydrogen peroxide to indirectly evaluate the oxidative stress in chicken liver. Results indicated that the oral administration of Cr(3+) at high dose significantly increased (P < 0.05) the MDA levels after 28 days of exposure, with decreased T-AOC, glutathione, and antioxidant enzymes activities. Low and medium doses groups show that T-AOC, glutathione, and antioxidant enzymes activities increased after 14 days, then decreased gradually, but low and medium groups higher than control group, only high group lower than control group finally. These statistics and histopathological analysis suggest that high dose and long-term exposure of Cr(3+) induce oxidative stress and hepatocellular injury.

The objective of this study is to evaluate the effects of chromic chloride (CrCl3) on chick embryo fibroblast (CEF) viability. The cells were incubated with CrCl3 (0.02, 0.1, 0.5, 2.5, 12.5, and 62.5 μM), and the viability was determined using MTT assay, morphological detection and flow cytometry. The results show that lower concentrations of CrCl3 (0.02, 0.1, and 0.5 μM) did not damage CEF viability. At 0.1 μM, CrCl3 can increase CEF viability (P

Poliomyelitis, a disease which can manifest as muscle paralysis, is caused by the poliovirus, which is a human enterovirus and member of the family Picornaviridae that usually transmits by the faecal–oral route. The viruses of the OPV (oral poliovirus attenuated-live vaccine) strains can mutate in the human intestine during replication and some of these mutations can lead to the recovery of serious neurovirulence. Informatics research of the poliovirus genome can be used to explain further the characteristics of this virus. In this study, sequences from 100 poliovirus isolates were acquired from GenBank. To determine the evolutionary relationship between the strains, we compared and analysed the sequences of the complete poliovirus genome and the VP1 region. The reconstructed phylogenetic trees for the complete sequences and the VP1 sequences were both divided into two branches, indicating that the genetic relationships of the whole poliovirus genome and the VP1 sequences are very similar. This branching indicates that the virulence and pathogenicity of poliomyelitis may be associated with the VP1 region. Sequence alignment of the VP1 region revealed numerous mutation sites in which mutation rates of >30 % were detected. In a group of strains recorded in the USA, mutation sites and mutation types were the same and this may be associated with their distribution in the evolutionary tree and their genetic relationship. In conclusion, the genetic evolutionary relationships of poliovirus isolate sequences are determined to a great extent by the VP1 protein, and poliovirus strains located on the same branch of the phylogenetic tree contain the same mutation spots and mutation types. Hence, the genetic characteristics of the VP1 region in the poliovirus genome should be analysed to identify the transmission route of poliovirus and provide the basis of viral immunity development.

Poliovirus, the causative agent of poliomyelitis, is a human enterovirus and member of the Picornaviridae family. An effective live-attenuated poliovirus vaccine strain (Sabin 1) has been developed and has protected humans from polio. However, a few cases of vaccine virulence reversion have been documented in several countries. For instance, circulating type 1 vaccine-derived poliovirus is a highly pathogenic poliovirus that evolved from an avirulent strain, but the mechanism by which vaccine strains undergo reversion remains unclear. In this study, vaccine strains exhibited A to G/U to C and G to A/C to U hypermutations in the reversed evolution of Sabin 1. Furthermore, the mutation ratios of U to C and C to U were higher than those of other mutation types. Dinucleotide editing context was then analyzed. Results showed that A to G and U to C mutations exhibited preferences similar to adenosine deaminases acting on RNA (ADAR). Hence, ADARs may participate in poliovirus vaccine evolution.

Klebsiella pneumoniae, which exists in the intestinal and respiratory tracts of humans and animals, is an important conditional pathogen in many animals. The aim of the current study was to investigate the antimicrobial resistance profiles and genotypes of extended-spectrum β-lactamase (ESBL)- and AmpC β-lactamase-producing K. pneumoniae isolated from dogs.A total of 285 isolates, collected from faecal and urine samples of diseased dogs in a Veterinary Teaching Hospital in Beijing, were characterised by antimicrobial susceptibility testing and screened for ESBL and AmpC β-lactamase phenotypes. The relevant genes were identified by polymerase chain reaction and sequencing.All K. pneumoniae isolates were susceptible to meropenem, while the rates of resistance to the remaining 27 tested antimicrobials ranged from 24% to 97%. A total of 53% and 18% of K. pneumoniae isolates were positive for ESBL and AmpC β-lactamase production, respectively. ESBL/AmpC-producing strains were significantly resistant to more antimicrobial agents compared non-ESBL/AmpC-producing strains (P0.05). CTX-M groups 1 and 9, and DHA-1 were the predominant genotypes of the ESBL/AmpC-producing K. pneumoniae isolates.In conclusion, the high percentage of drug resistance among K. pneumoniae isolates suggests that routine detection of ESBL production by reliable laboratory methods is required in small animal clinical practice.

Clostridium butyricum (C. butyricum) can attenuate oxidative stress, inflammation, and hepatic fatty deposition in poultry, however, the underlying mechanisms for this in Pekin ducks remain unclear. This study evaluated these hepatoprotective effects and the underlying mechanisms in a corticosterone (CORT)-induced liver injury model in Pekin ducks fed a C. butyricum intervention diet. A total of 500 Pekin ducks were randomly divided into five groups: one group (CON group) was only provided with a basal diet, three groups were provided a basal diet with 200 mg/kg (LCB group), 400 mg/kg (MCB group), or 600 mg/kg (HCB group) C. butyricum, respectively, and one group was provided a basal diet with 150 mg/kg aureomycin (ANT group) for 42 d. At 37 days-old, all ducks received daily intraperitoneal injections of CORT for five days to establish a liver injury model. C. butyricum intervention alleviated liver injury by decreasing the liver organ indices, hepatic steatosis and hepatocyte necrosis, and improving liver function, antioxidant capacity, and inflammatory factors. Hepatic RNA-seq revealed 365 differentially expressed genes (DEGs) between the MCB and CON groups, with 229 up- and 136 down-regulated DEGs in the MCB group. Between the MCB and ANT groups, 407 DEGs were identified, including 299 up- and 108 down-regulated genes in MCB group. Some DEGs in the MCB group related to oxidative stress and inflammatory responses such as Sod3, Tlr2a/b, and Il10, which were up-regulated, while Apoa1, Cyp7a1, Acsl1/5, Fasn, Ppar-&gamma, and Scd, which are involved in lipid metabolism, were down-regulated, indicating that these genes were responsive to dietary C. butyricum for the alleviation of corticosterone-induced hepatic injury. Toll-like receptor signaling, PI3K-Akt signaling pathway, cytokine-cytokine receptor interaction, peroxisome proliferator-activated receptor (PPAR) signaling pathway, adipocytokine and glycerophospholipid metabolism signaling pathway were significantly enriched in the MCB group. These findings indicate that C. butyricum intervention can protect Pekin ducks from corticosterone-induced liver injury by the modulation of immunoregulatory- and lipid metabolism-related genes and pathways.

This study aimed to investigate the effects of chromic chloride (CrCl

This study aimed to evaluate the effects of adding chromic chloride (CrCl3) in the drinking water of chickens. Hyland brown male chickens were randomly divided into four groups. Three groups orally received 1/2 LD50, 1/4 LD50, and 1/8 LD50 CrCl3mgkg(-1) body weight daily for 42d. The fourth group was treated with water. The chickens were sacrificed at 14, 28, and 42d post-treatment. The renal injury was examined through histological analysis, and kidney mass was determined. The effects on growth performance were assessed by measuring the weight of the body, chest muscles, and leg muscles. Oxidative damage was evaluated by determining the antioxidant defense levels in kidney homogenates. The body weight and the weight of tissues gained time-dependently, but significantly decreased compared with those in the control group (P

Low molecular weight polypeptides 2 (LMP2) and low molecular weight polypeptides 7 (LMP7) are located within the major histocompatibility complex and have been associated with autoimmune disease. In this study, polymorphisms of porcine LMP2 and LMP7 genes were analyzed by PCR-SSCP and DNA sequencing methods. Four SNPs (DQ659151:g.2115T>C; DQ659151:g.4343A>G; DQ872631:g.1232C>G; DQ872631:g.2847C>T) were identified. Four SNPs of genes were analyzed for association with 22 haematological traits in Large White (n = 195), Landrace (n = 84) and Songliao Black (n = 86) pig population. Of all the 22 traits, seven were significant associated with the SNPs of LMP2/LMP7 gene (P < 0.05). They included white blood cell count (WBC) (P = 0.028), neutrophilic granulocyte count (GRAN) (P = 0.037), monocytes percentage (MO%) (P = 0.015), red blood cell (RBC) (P = 0.004), red blood cell volume distribution width (RDW) (P = 0.004), mean platelet volume (MPV) (P = 0.016) and CD4(+)CD8(+)% (P = 0.045). These results suggest LMP2/LMP7 gene should be regarded as molecular marker to estimate animal's immune status for their effects on hematological traits.

Interferon regulatory factor 1 (IRF1) is a member of IRF-family that was discovered to activate promoters in type I interferon (IFN) genes. It is shown to play functionally diverse role in the regulation of the immune system. In this report, the porcine IRF1 cDNA were cloned and a 7500 bp genomic DNA structure was identified. The putative IRF1 protein included 322 amino acids. Alignment and phylogenetic analysis of the predicted porcine IRF1 amino acids sequence with its homologies of other species show high identity (over 88%). Tissues expression of IRF1 mRNA was observed by RT-PCR, the results revealed IRF1 gene expressed widely in all analyzed tissues. Using the radiation hybrid panel, the porcine IRF1 gene was mapped to porcine chromosome 2 and closely linked to the locus IL4 (LOD = 7.09, 57cR). A SNP in exon2 of porcine IRF1 gene was demonstrated by sequencing and PCR–RFLP analysis. The further association analysis indicated that the SNP was significant associate with level of IFN-γ (day 20) in serum (P = 0.0001) and the ratio of IFN-γ to IL10 (day 20; day 35) in serum (P = 0.0165; P = 0.0095). The results suggested that the porcine IRF1 gene is strong candidate gene for these immune traits in pig.