Investigating the genetic architecture of disease resilience in pigs by genome-wide association studies of complete blood count traits collected from a natural disease challenge model.

BACKGROUND: Genetic improvement for disease resilience is anticipated to be a practical method to improve efficiency and profitability of the pig industry, as resilient pigs maintain a relatively undepressed level of performance in the face of infection. However, multiple biological functions are known to be involved in disease resilience and this complexity means that the genetic architecture of disease resilience remains largely unknown. Here, we conducted genome-wide association studies (GWAS) of 465,910 autosomal SNPs for complete blood count (CBC) traits that are important in an animal's disease response. The aim was to identify the genetic control of disease resilience. RESULTS: Univariate and multivariate single-step GWAS were performed on 15 CBC traits measured from the blood samples of 2743 crossbred (Landrace × Yorkshire) barrows drawn at 2-weeks before, and at 2 and 6-weeks after exposure to a polymicrobial infectious challenge. Overall, at a genome-wise false discovery rate of 0.05, five genomic regions located on Sus scrofa chromosome (SSC) 2, SSC4, SSC9, SSC10, and SSC12, were significantly associated with white blood cell traits in response to the polymicrobial challenge, and nine genomic regions on multiple chromosomes (SSC1, SSC4, SSC5, SSC6, SSC8, SSC9, SSC11, SSC12, SSC17) were significantly associated with red blood cell and platelet traits collected before and after exposure to the challenge. By functional enrichment analyses using Ingenuity Pathway Analysis (IPA) and literature review of previous CBC studies, candidate genes located nearby significant single-nucleotide polymorphisms were found to be involved in immune response, hematopoiesis, red blood cell morphology, and platelet aggregation. CONCLUSIONS: This study helps to improve our understanding of the genetic basis of CBC traits collected before and after exposure to a polymicrobial infectious challenge and provides a step forward to improve disease resilience.

Hepatitis E virus prevalence among blood donors in Dali, China.

BACKGROUND: Hepatitis E virus (HEV) is a nonenveloped RNA virus causing hepatitis E worldwide. The increase in transfusion-transmitted cases of HEV infections from asymptomatic blood donors causing serious illnesses among immunosuppressed recipients has been reported in the past few years. China is one of the most prevalent regions of HEV; as a result, it is important to evaluate the risk of transfusion-transmitted HEV. METHODS: A total of 1864 serum samples (including demographic characteristics) from blood donors were randomly collected from February to March 2018 in Dali city. Anti-HEV IgG, IgM and IgA antibodies and HEV antigen were examined by enzyme-linked immunosorbent assay (ELISA). HEV RNA was detected by real-time PCR. Multivariable logistic regression modelling was used to examine the risk factors associated with HEV prevalence. RESULTS: Overall, the positive rates of anti-HEV IgG, IgM, and IgA antibodies were 13.36% (249/1864), 1.13% (21/1864), and 1.82% (34/1864), respectively. However, none of the 1864 serum samples were HEV antigen positive or HEV RNA positive. Females (16.69%) had a significantly higher HEV seroprevalence than males (13.04%) (odds ratio [OR] 1.34 [95% CI, 1.02-1.75]). Bai (18.85%) donors had a significantly higher HEV seroprevalence than Han (12.21%) blood donors (odds ratio [OR], 1.65 [95% CI, 1.24-2.19] for Bai). CONCLUSIONS: HEV showed a seroprevalence among blood donors in Yunnan Province, some of which were even recent infections, indicating a threat to the safety of blood transfusions. Whether to formulate a strategy for HEV screening in blood centres needs further research.

Genome-wide association study identifies candidate genes for piglet splay leg syndrome in different populations.

BACKGROUND: Piglet splay leg syndrome (PSL) is one of the most frequent genetic defects, and can cause considerable economic loss in pig production. The present understanding of etiology and pathogenesis of PSL is poor. The current study focused on identifying loci associated with PSL through a genome-wide association study (GWAS) performed with the Illumina Porcine60 SNP Beadchip v2.0. The study was a case/control design with four pig populations (Duroc, Landrace, Yorkshire and one crossbred of Landrace × Yorkshire). RESULT: After quality control of the genotyping data, 185 animals (73 cases, 112 controls) and 43,495 SNPs were retained for further analysis. Principal components (PCs) identified from the genomic kinship matrix were included in the statistical model for correcting the effect of population structure. Seven chromosome-wide significant SNPs were identified on Sus scrofa chromosome 1 (SSC1), SSC2 (2 SNPs), SSC7, SSC15 (2 SNPs) and SSC16 after strict Bonferroni correction. Four genes (HOMER1 and JMY on SSC2, ITGA1 on SSC16, and RAB32 on SSC1) related to muscle development, glycogen metabolism and mitochondrial dynamics were identified as potential candidate genes for PSL. CONCLUSIONS: We identified seven chromosome-wide significant SNPs associated with PSL and four potential candidate genes for PSL. To our knowledge, this is the first pilot study aiming to identify the loci associated with PSL using GWAS. Further investigations and validations for those findings are encouraged.

Iteratively reweighted LASSO for mapping multiple quantitative trait loci.

The iteratively reweighted least square (IRLS) method is mostly identical to maximum likelihood (ML) method in terms of parameter estimation and power of quantitative trait locus (QTL) detection. But the IRLS is greatly superior to ML in terms of computing speed and the robustness of parameter estimation. In conjunction with the priors of parameters, ML can analyze multiple QTL model based on Bayesian theory, whereas under a single QTL model, IRLS has very limited statistical power to detect multiple QTLs. In this study, we proposed the iteratively reweighted least absolute shrinkage and selection operator (IRLASSO) for extending IRLS to simultaneously map multiple QTLs. The LASSO with coordinate descent step is employed to efficiently estimate non-zero genetic effect of each locus scanned over entire genome. Simulations demonstrate that IRLASSO has a higher precision of parameter estimation and power to detect QTL than IRLS, and is able to estimate residual variance more accurately than the unweighted LASSO based on LS. Especially, IRLASSO is very fast, usually taking less than five iterations to converge. The barley dataset from the North American Barley Genome Mapping Project is reanalyzed by our proposed method.

Sanders II-III calcaneal fractures fixed with locking plate in elderly patients.

PURPOSE: To evaluate the clinical outcomes of locking calcaneal plate in treating calcaneal fracture (Sanders II-III) in elderly patients. METHODS: From October 2012 to December 2013, 23 elderly patients suffering from calcaneal fracture (Sanders II-III) were treated and followed up. There were 15 males and 8 females with the mean age of 68.5 years (range: 65-79 years). According to Sander's classification, 16 cases (16 feet) were type II fractures and 7 cases (7 feet) were type III fractures. Anteroposterior, lateral and axial views of X-ray were taken to detect the calcaneum. CT scan was done to assess the amount of comminution and articular depression. Radiological assessment was performed using Bohler's angle and Gissane's angle. Functional outcome was assessed using the Maryland foot score. RESULTS: All the patients were followed up for 13.7 months on average (10-20 months). The mean time of bone union was 3.2 months (3-4 months). The mean time of complete weight bearing was 3.2 months (3.1-4.0 months). The soft tissue necrosis was found in 1 case. The mean Bohler's angle and Gissane's angle were 25.31° and 117.5°respectively. The overall excellent to good rate was 82.6%. CONCLUSION: Open reduction and internal fixation with locking calcaneal plate can obtain good functional outcome for Sanders II-III calcaneal fractures in elderly patients.

Genome-wide association studies (GWAS) identify a QTL close to PRKAG3 affecting meat pH and colour in crossbred commercial pigs.

BACKGROUND: Improving meat quality is a high priority for the pork industry to satisfy consumers' preferences. GWAS have become a state-of-the-art approach to genetically improve economically important traits. However, GWAS focused on pork quality are still relatively rare. RESULTS: Six genomic regions were shown to affect loin pH and Minolta colour a* and b* on both loin and ham through GWAS in 1943 crossbred commercial pigs. Five of them, located on Sus scrofa chromosome (SSC) 1, SSC5, SSC9, SSC16 and SSCX, were associated with meat colour. However, the most promising region was detected on SSC15 spanning 133-134 Mb which explained 3.51% - 17.06% of genetic variance for five measurements of pH and colour. Three SNPs (ASGA0070625, MARC0083357 and MARC0039273) in very strong LD were considered most likely to account for the effects in this region. ASGA0070625 is located in intron 2 of ZNF142, and the other two markers are close to PRKAG3, STK36, TTLL7 and CDK5R2. After fitting MARC0083357 (the closest SNP to PRKAG3) as a fixed factor, six SNPs still remained significant for at least one trait. Four of them are intragenic with ARPC2, TMBIM1, NRAMP1 and VIL1, while the remaining two are close to RUFY4 and CDK5R2. The gene network constructed demonstrated strong connections of these genes with two major hubs of PRKAG3 and UBC in the super-pathways of cell-to-cell signaling and interaction, cellular function and maintenance. All these pathways play important roles in maintaining the integral architecture and functionality of muscle cells facing the dramatic changes that occur after exsanguination, which is in agreement with the GWAS results found in this study. CONCLUSIONS: There may be other markers and/or genes in this region besides PRKAG3 that have an important effect on pH and colour. The potential markers and their interactions with PRKAG3 require further investigation.

Roles of leptin in bone metabolism and bone diseases.

Adipose tissue has been more accepted as an active contributor to whole body homeostasis, rather than just a fat depot, since leptin, a 16 kDa protein, was discovered as the product of the obese gene in 1994. With more and more studies conducted on this hormone, it has been shown that there is a close relationship between adipose tissue and bone, which have important effects on each other. Bone is the source of many hormones, such as osteocalcin, that can affect energy metabolism and then the anabolism or catabolism of fat tissue. In contrast, the adipose tissue synthesizes and releases a series of adipokines, which are involved in bone metabolism through direct or indirect effects on bone formation and resorption. Interestingly, leptin, one of the most important cytokines derived from fat tissue, seems to account for the largest part of effects on bone, through direct or indirect involvement in bone remodeling and by playing a significant role in many bone diseases, such as osteoporosis, osteoarthritis, rheumatic arthritis, bone tumors and even fractures. In this review, we will discuss the progress in leptin research, particularly focusing on the roles of leptin in bone diseases.

The evaluation of patient-specific factors associated with meniscal and chondral injuries accompanying ACL rupture in young adult patients.

PURPOSE: To evaluate patient-specific factors, including the mechanism of injury, time from the injury, activity level after the initial trauma, re-injury and body mass index (BMI), as risk factors for meniscal and chondral injuries accompanying anterior cruciate ligament (ACL) rupture. METHODS: A retrospective review of young adult patients with complete ACL rupture was performed between 2007 and 2011. The presence of meniscal tears and/or chondral injury, and the mechanism of injury, time from the injury, activity level after the initial trauma, re-injury and BMI were recorded. The data were analysed for associations between patient-specific factors and the prevalence of meniscal and chondral injuries. RESULTS: A total of 293 patients were included. Increasing time from the injury, active daily life and re-injury were risk factors associated with meniscal injuries (p < 0.05). Independent risk factors associated with meniscal injuries included active daily life (OR = 4.66, 95 % CI 2.21-9.86, p < 0.0001) and re-injury (OR = 7.68, 95 % CI 3.24-18.22, p < 0.0001). Contact injury, increasing time from the injury, active daily life and re-injury were risk factors associated with chondral injuries (p < 0.05). Independent risk factors associated with chondral injuries included contact injury (OR = 2.54, 95 % CI 1.27-5.10, p < 0.01), active daily life (OR = 2.62, 95 % CI 1.35-5.08, p < 0.01) and re-injury (OR = 4.18, 95 % CI 2.09-8.35, p < 0.001). The incidence of re-injury was associated with the activity level (p < 0.001) and increased with increasing time from the injury (r = -0.698, p < 0.001). CONCLUSIONS: An increased risk of meniscal tear (especially medial meniscus) was strongly associated with an active daily life and re-injury. The combination of contact injury, active daily life and re-injury substantially increased the risk of cartilage injury. Patients with increasing time from the injury and active daily life exhibited a higher risk of re-injury, implying a higher prevalence of intra-articular damage.

Comparison of three plate system for lateral malleolar fixation.

BACKGROUND: This study was to compare clinical and radiographic outcomes with three different implants and evaluate the effectiveness of minimally invasive plate osteosynthesis (MIPO) technique for the distal fibular fractures. METHODS: We performed a retrospective cohort single-surgical team single-facility study between 2000 and 2011. 147 patients receiving surgical interventions for closed, displaced distal fibular fractures were included. Based on the different implants, patients were divided into three groups: Group A: one-third tubular plate; Group B: locking compression (LCP) metaphyseal plate; Group C: LCP distal fibula plate. Clinical and radiographic outcomes were compared among the three groups. RESULTS: Totally, we found that patients in Group C had significant higher functional scores than those in Group A (p1 = 0.004; p2 = 0.002) (p1 stands for the p value for Olerud & Molandar Score, p2 stands for the p value for American Orthopaedic Foot & Ankle Society score). The healing time was significant less in Group C than that in Group A (p < 0.0001) and Group B (p < 0.0001). Subgroup analysis showed that: (1) For Weber A fracture, the functional scores of the Group C were higher than those in Group A (p1 = 0.020; p2 = 0.029) and B (p1 = 0.020; p2 = 0.034). (2) For Weber B fracture, the functional scores of the Group B (p1 = 0.033; p2 = 0.030) and C (p1 = 0.027; p2 = 0.017) were higher than those in Group A. No significant differences were observed in terms of the ankle range of motion, reduction accuracy and complication rate. CONCLUSIONS: Our study demonstrated using LCP metaphyseal plate in patients associated with lateral malleolar fracture could achieve significantly better OMS & AOFAS scores and less healing time than using one-third tubular plate. Specifically, For Weber A fracture, LCP distal fibula plate is much better than one-third tubular plate and LCP metaphyseal plate. While for Weber B fracture, LCP distal fibula plate and LCP metaphyseal plate are better than one-third tubular plate. As to the complications, using MIPO technique in patients with distal fibular fractures is at least comparable to the traditional one.

Effect of dynamic hip screw on the treatment of femoral neck fracture in the elderly.

OBJECTIVE: To discuss the indications, surgical procedures, and curative effect of dynamic hip screw (DHS) in the treatment of femoral neck fracture in the elderly. METHODS: A retrospective study was conducted to analyse the clinical data of 42 elderly patients who had been treated for femoral neck fracture with DHS in our department between June 2009 and November 2011. There were 21 males and 21 females with a mean age of 68.5 years (range 60-75 years). According to the Garden Classification, there were 19 cases of type II, 21 cases of type III and 2 cases of type IV fractures. By the Singh Index Classification, there were 3 cases of level 2, 19 cases of level 3 and 20 cases of level 4 fractures. The Harris criterion, complications and function recovery after opera- tion were analysed. RESULTS: The average hospitalization time in 42 patients was 11.2 days (range 7-21 days). All patients were followed up for 12-26 months (mean 18 months). No lung infection, deep venous thrombosis or other complications occurred. Partial backing-out of the screws was found in 2 cases. The internal fixation device was withdrawn after fracture healing. Internal fixation cutting was found in 1 case, and he had a good recovery after total hip arthroplasty. The time for fracture healing ranged from 3-6 months (average 4.5 months). According to Harris criterion, 15 cases were rated as excellent, 24 good, 2 fair and 1 poor. The Harris scale was significantly improved from 30.52±2.71 preoperatively to 86.61±2.53 at 6 months postoperatively (P<0.05). CONCLUSION: DHS, being minimal invasive, allowing early activity and weight-bearing, is advisable for treatment of elderly patients with femoral neck fracture. In addition, it can avoid complications seen in artificial joint replacement. It is especially suitable for patients with mild osteoporosis.

Rapid enrichment of AnAOB with a novel vermiculite/tourmaline modification technology for enhanced DEAMOX process.

The DEnitrifying AMmonium OXidation (DEAMOX) has been proven to be a promising process treating contaminated surface water containing ammonia and nitrate, while the enrichment of the slow-growing anammox bacteria (AnAOB) remains a challenge. In this study, a novel polyurethane-adhesion vermiculite/tourmaline (VTP) modified carrier was developed to achieve effective enrichment of AnAOB. The results demonstrated that the VTP-1 (vermiculite: tourmaline = 1:1) system exhibited the greatest performance with the total nitrogen removal efficiency reaching 87.6% and anammox contributing 63% to nitrogen removal. Scanning electron microscope analysis revealed the superior biofilm structure of the VTP-1 carrier, providing attachment for AnAOB. The addition of VTP-1 promoted the secretion of EPS (extracellular polymeric substances) by microorganisms, which increased to 85.34 mg/g VSS, contributing to the aggregation of anammox cells. The favorable substrate microenvironment created by NH4+ adsorption and NO2- supply via partial denitrification process facilitated the growth of AnAOB. The relative abundance of Candidatus Brocadia and Thauera increased from 0.04% to 0.3%-1.03% and 2.06% in the VTP-1 system, respectively. This study sheds new light on the anammox biofilm formation and provides a valid approach to initiate the DEAMOX process for low nitrogen polluted water treatment.

Management of limb fractures in a teaching hospital: comparison between Wenchuan and Yushu earthquakes.

OBJECTIVE: To comparatively analyze the medical records of patients with limb fractures as well as rescue strategy in Wenchuan and Yushu earthquakes so as to provide references for post-earthquake rescue. METHODS: We retrospectively investigated 944 patients sustaining limb fractures, including 891 in Wenchuan earthquake and 53 in Yushu earthquake, who were admitted to West China Hospital (WCH) of Sichuan University. RESULTS: In Wenchuan earthquake, WCH met its three peaks of limb fracture patients influx, on post-earthquake day (PED) 2, 8 and 14 respectively. Between PED 3-14, 585 patients were transferred from WCH to other hospitals outside the Sichuan Province. In Yushu earthquake, the maximum influx of limb fracture patients happened on PED 3, and no one was shifted to other hospitals. Both in Wenchuan and Yushu earthquakes, most limb fractures were caused by blunt strike and crush/burying. In Wenchuan earthquake, there were 396 (396/942, 42.0%) open limb fractures, including 28 Gustilo I, 201 Gustilo II and 167 Gustilo III injuries. But in Yushu earthquake, the incidence of open limb fracture was much lower (6/61, 9.8%). The percent of patients with acute complications in Wenchuan earthquake (167/891, 18.7%) was much higher than that in Yushu earthquake (5/53, 3.8%). In Wenchuan earthquake rescue, 1 018 surgeries were done, composed of debridement in 376, internal fixation in 283, external fixation in 119, and vacuum sealing drainage in 117, etc. While among the 64 surgeries in Yushu earthquake rescue, the internal fixation for limb fracture was mostly adopted. All patients received proper treatment and survived except one who died due to multiple organs failure in Wenchuan earthquake. CONCLUSION: Provision of suitable and sufficient medical care in a catastrophe can only be achieved by construction of sophisticated national disaster medical system, prediction of the injury types and number of injuries, and confirmation of participating hospitals?exact role. Based on the valuable rescue experiences after Wenchuan earthquake, the rescue was faster, more orderly and effective in Yushu earthquake. Nevertheless, there is still a long way to go in the development of a stronger emergent response to the disasters.

[Comparative analysis of the clinical characteristics of orthopedic inpatients in Lushan and Wenchuan earthquakes].

OBJECTIVE: To systematically analyze and compare the clinical characteristics of orthopedic inpatients in Lushan and Wenchuan earthquake, so as to provide useful references for future earthquakes injury rescue. METHODS: Based on the orthopedic inpatients in Lushan and Wenchuan earthquakes, the data of the age, gender, injury causes, body injured parts and speed of transport were classified and compared. RESULTS: The duration of patients admitted to hospital lasted long and the peak appeared late in Wenchuan earthquake, which is totally opposed to Lushan earthquake. There was no significant difference in the patient's age and gender between the two earthquakes. However, the occurrence rate of crush syndrome, amputation, gas gangrene, vascular injury and multiple organ dysfunction syndrome (MODS) in Wenchuan earthquake was much higher than that in Lushan earthquake. Blunt traumas or crush-related injuries (79.6%) are the major injury cause in Wenchuan earthquake, however, high falling injuries and falls (56.8%) are much higher than blunt trauma or crush-related injuries (39.2%) in Lushan earthquake. The incidence rate of foot fractures, spine fractures and multiple fractures in Lushan earthquake was higher than that in Wenchuan earthquake, but that of open fractures and lower limb fractures was lower than that in Wenchuan earthquake. CONCLUSION: The rapid rescue scene is the cornerstone of successful treatment, early rescue and transport obviously reduce the incidence of the wound infection, crush syndrome, MODS and amputation. Popularization of correct knowledge of emergency shelters will help to reduce the damage caused by blindly jumping or escaping while earthquake happens.

Design strategies of covalent organic framework-based electrodes for supercapacitor application.

Supercapacitors (SCs) have been recognized as a promising electrochemical energy storage (EES) device, thanks to their high-power density, long lifespan, fast charge-discharge capability, and eco-friendliness. The breakthrough of electrode materials that determine the electrochemical performance of SCs is urgently desired. Covalent organic frameworks (COFs), an emerging and burgeoning class of crystalline porous polymeric materials, have been found to have huge potential for application in EES devices by virtue of their unique properties including atomically adjustable structures, robust and tunable skeletons, well-defined and open channels, high surface areas, etc. In this feature article, we aim at summarizing the design strategies of COF-based electrode materials for SCs based on the representative advances. The current challenges and future perspectives of COFs for SC application are highlighted as well.

The biochar/Fe-modified biocarrier driven simultaneous NDFO and Feammox to remove nitrogen from eutrophic water.

Novelty techniques of Fe(III) reduction coupled to anaerobic ammonium oxidation (i.e. Feammox) and nitrate-dependent Fe(II) oxidation (i.e. NDFO) provide new insights into autotrophic nitrogen removal from eutrophic waters. Given that Feammox and NDFO can theoretically complete the simultaneous NH+ 4-N and NO- 3-N removal via Fe(III)/Fe(II) cycle, this study introduces iron powder to the surface of the biocarrier as a solid-phase source of Fe, and biochar was used as an electron shuttle to mix with the iron powder to improve the bioavailability of iron. Batch experiments was carried out for 70 days using simulated eutrophic water as the medium to investigate the effects of the modified biocarrier for enhanced nitrogen removal. The results showed that BC1 (Fe:BC=1:1) with the highest relative Fe content exhibited the highest nitrogen removal efficiency of 66.74%. XPS and XRD results showed both Fe(III) and Fe(II) compounds on the biocarrier surface, confirming the occurrence of Fe(III)/Fe(II) cycle. The ex-situ activity test indicated that functional activity was positively correlated with the iron content of the biocarrier. The in-situ experiments with different substrates showed the occurrence of Feammox and NDFO. NDFO bacteria (Gallionellaceae), Feammox bacteria (Alicycliphilus), denitrifying and digesting bacteria were enriched, suggesting that the coupled nitrogen removal of NDFO and Feammox is the result of cooperation between different functional microorganisms. Thus, the Fe-modified biocarrier showed superior performance and application potential in catalyzing autotrophic nitrogen removal from eutrophic water by functional microorganisms.

Bayesian mapping of genome-wide epistatic imprinted loci for quantitative traits.

Genomic imprinting, an epigenetic phenomenon of parent-of-origin-specific gene expression, has been widely observed in plants, animals, and humans. To detect imprinting genes influencing quantitative traits, the least squares and maximum likelihood approaches for fitting a single quantitative trait locus (QTL) and Bayesian methods for simultaneously modeling multiple QTL have been adopted, respectively, in various studies. However, most of these studies have only estimated imprinting main effects and thus ignored imprinting epistatic effects. In the presence of extremely complex genomic imprinting architectures, we introduce a Bayesian model selection method to analyze the multiple interacting imprinted QTL (iQTL) model. This approach will greatly enhance the computational efficiency through setting the upper bound of the number of QTLs and performing selective sampling for QTL parameters. The imprinting types of detected main-effect QTLs can be estimated from the Bayes factor statistic formulated by the posterior probabilities for the genetic effects being compared. The performance of the proposed method is demonstrated by several simulation experiments. Moreover, this method is applied to dissect the imprinting genetic architecture for body weight in mouse and fruit weight in tomato. Matlab code for implementing this approach will be available from the authors upon request.

Overexpression of GDF5 through an adenovirus vector stimulates osteogenesis of human mesenchymal stem cells in vitro and in vivo.

The use of stem cells combined with gene therapy could be an important way to facilitate bone regeneration. In this study, the aim was to investigate the potential of growth and differentiation factor-5 (GDF5) to genetically manipulate human mesenchymal stem cells (hMSCs) for bone regeneration. Recombinant adenovirus Ad-GDF5 and Ad-GFP were constructed and identified, and the titer of both were determined. Third-passage hMSCs were infected with adenovirus, and the expression of GDF5 was confirmed by detection of GFP-positive cells, GDF5 mRNA levels, Western blotting, and enzyme-linked immunosorbent assay (ELISA). hMSCs at passage 3 were divided into four groups: (1) an experimental group infected with Ad-GDF5, (2) a positive control group cultured with osteogenic differentiation medium, (3) a control group infected with Ad-GFP cultured with standard medium, and (4) a blank control group cultured with standard medium. Evaluation of cell morphology and proliferation, analysis of the expression of genes related to osteogenic differentiation, von Kossa staining, and immunofluorescent staining of collagen I were used to investigate the osteogenesis of cells among the groups. After culturing the cells for 2 days under each corresponding condition, the cells were detached and subcutaneously injected into the backs of nude mice to evaluate bone formation. Samples were collected for histological staining, protein Western blotting, and micro-computer tomography. When infected with Ad-GDF5, hMSCs could overexpress GDF5 for a prolonged period in vitro and reach a concentration of 160 ng/ml. Cells infected with Ad-GDF5 or cultured in osteogenic medium displayed osteogenic differentiation based on their histological and cellular properties and on their gene and protein expression patterns. Furthermore, Ad-GDF5 showed a better ability to upregulate the expression of collagen I, alkaline phosphatase, and osteocalcin mRNA than the osteogenic medium. Furthermore, Ad-GDF5 expression was associated with enhanced bone formation in vivo. Our findings suggest that hMSCs infected with Ad-GDF5 can differentiate in an osteogenic direction and may be a promising cell source for bone regeneration.

Timing of internal fixation and effect on Schatzker IV-VI tibial plateau fractures.

OBJECTIVE: To study the effect of internal fixation performed at different times on therapeutic outcomes of Schatzker IV-VI tibial plateau fractures. METHODS: The clinical data of 42 cases of Schatzker IV-VI tibial plateau fractures treated in our department were analyzed retrospectively. Among these 42 patients, 21 received surgical treatment within 12 h after injury (Group I), the other 21 were first treated by traction or plaster fixation followed by a delayed internal fixation after soft tissue swelling subsided (Group II). The surgical time, complications, length of hospital stay, cost of hospitalization, and time for fracture union, as well as functional recovery were analyzed and compared between the two groups. RESULTS: After 10-28 months follow-up (mean 16.5 months), except 5 cases who lost to follow-up, no differences were found between the two groups regarding surgical time, preoperative and postoperative complications, healing time or the Hospital for Special Surgery (HSS) score at the end of follow-up, but significant differences were found in the length of hospital stay, cost of hospitalization and HSS score at 3 months after operation (P less than 0.05). CONCLUSION: Under certain conditions, early internal fixation for Schatzker IV-VI tibial plateau fracture is feasible, which can shorten the length of hospital stay, decrease the cost of hospitalization and promote early functional rehabilitation.

[Screen and identification of the relative protein expressed in the acute tractive spinal cord injury in rats].

OBJECTIVE: To screen and identify the relative protein expressed in the acute tractive spinal cord injury (TSCI) in rats. METHODS: Ten adult Sprague Dawley rats were randomly divided into Sham group and TSCI group, 5 rats in each group. Rats from Sham group and TSCI group at 1 day after surgery were sacrificed for harvesting T13-L2 spinal tissue specimens. The extraction and quantitation of protein in the spinal tissue was finished firstly. Proteins from spinal tissue were separated by two-dimensional electrophoresis (2-DE) and identified by mass spectrometry (MS). The different expression map was established in each group, and proteins express differently was determined by comparing the level of each spot with gel imaging software and manually. Proteins were identified by High performance liquid chromatography-electrospray tandem (NanoUPLC-ESI-MS/MS) and peptide sequence tag with tandem MS combining with database respectively. After that, the function of these identified proteins was known and classified. RESULTS: There were 22 differential protein expression spots were found between Sham group and TSCI group. Among them, 18 spots were up-regulated and 4 were down-regulated. 4 differential protein expression spots were newly found in TSCI group. Sixteen significant proteins were identified by NanoUPLC-ESI-MS/MS. Four kind of proteins were related to apoptosis, 3 in nerve signal transduction and 6 in metabolism, respectively. Unnamed proteins were 3. CONCLUSIONS: The differential expression proteins were found between Sham group and TSCI group. These identified proteins may play important role in the process of injury and recovery through transduction nerve signal, regulating nerve cells apoptosis and metabolism.

Heritable and Nonheritable Rumen Bacteria Are Associated with Different Characters of Lactation Performance of Dairy Cows.

Recent studies have reported that some rumen microbes are heritable. However, it is necessary to clarify the functions and specific contributions of the heritable rumen microbes to cattle phenotypes (microbiability) in comparison with those that are nonheritable. This study aimed to identify the distribution and predicted functions of heritable and nonheritable bacterial taxa at species level in the rumen of dairy cows and their respective contributions to energy-corrected milk yield, protein content and yield, and fat content and yield in milk. Thirty-two heritable and 674 nonheritable bacterial taxa were identified at species level, and the functional analysis revealed that predicted microbial functions for both groups were mainly enriched for energy, amino acid, and ribonucleotide metabolism. The mean microbiability (to reflect a single taxon's contribution) of heritable bacteria was found to range from 0.16% to 0.33% for the different milk traits, whereas the range for nonheritable bacteria was 0.03% to 0.06%. These findings suggest a strong contribution by host genetics in shaping the rumen microbiota, which contribute significantly to milk production traits. Therefore, there is an opportunity to further improve milk production traits through attention to host genetics and the interaction with the rumen microbiota. IMPORTANCE Rumen bacteria produce volatile fatty acids which exert a far-reaching influence on hepatic metabolism, mammary gland metabolism, and animal production. In the current study, 32 heritable and 674 nonheritable bacterial taxa at species level were identified, and shown to have different microbiability (overall community contribution) and mean microbiability (the average of a single taxon's contribution) for lactation performance. The predicted functions of heritable and nonheritable bacterial taxa also differed, suggesting that targeted nutritional and genetic breeding approaches could be used to manipulate them to improve dairy cow performance.