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BackgroundMicrosatellite instability (MSI) is a biomarker for better outcomes in colorectal cancer (CRC). However, this conclusion is controversial. In addition, MSs can be a useful marker for loss of heterozygosity (LOH) of genes, but this finding has not been well studied. Here, we aimed to clarify the predictive value of MSI/LOH within tumor-related genes in CRC.MethodsWe detected MSI/LOH of MSs in tumor-related genes and the Bethesda (B5) panel by STR scanning and cloning/sequencing. We further analyzed the relationship between MSI/LOH status and clinical features or outcomes by Pearson’s Chi-square test, Fisher’s exact test and the Kaplan–Meier method.ResultsThe findings indicated that the MSI rates of B5 loci were all higher than those of loci in tumor-related genes. Interestingly, MSI/LOH of 2 loci in the B5 panel and 12 loci in tumor-related genes were associated with poorer outcomes, while MSI/LOH of the B5 panel failed to predict outcomes in CRC. MSI of BAT25, MSI/LOH of BAT26 and MSI of the B5 panel showed closer relationships with mucinous carcinoma. In addition, LOH-H of the B5 panel was associated with increased lymphatic metastasis.ConclusionsIn summary, MSI/LOH of certain loci or the whole panel of B5 is related to clinical features, and several loci within tumor-related genes showed prognostic value in the outcomes of CRC.

In this paper, ER50-6 steel was fabricated by wire arc additive manufacturing (WAAM) with an A-W GTAW system. The microstructure, mechanical properties, and corrosion behaviors of ER50-6 steel by WAAM were studied. The results showed that, with the GMAW current increased, from the bottom to the top of the sample, the microstructure was fine ferrite and granular pearlite, ferrite equiaxed grains with fine grains at grain boundaries, and columnar ferrite, respectively. The average hardness in the vertical direction of samples 1# and 2# was 146 and 153 HV, respectively. The hardness of sample 2# increased because of the refinement of grain. The pores in the sample increased as the bypass current increased. The higher bypass current also has a deterioration effect on the corrosion behavior of ER50-6 steel.

Unmanned autonomous helicopter (UAH) path planning problem is an important component of the UAH mission planning system. The performance of the automatic path planner determines the quality of the UAH flight path. Aiming to produce a high-quality flight path, a path planning system is designed based on human-computer hybrid augmented intelligence framework for the UAH in this paper. Firstly, an improved artificial bee colony (I-ABC) algorithm is proposed based on the dynamic evaluation selection strategy and the complex optimization method. In the I-ABC algorithm, the following way of on-looker bees and the update strategy of nectar source are optimized to accelerate the convergence rate and retain the exploration ability of the population. In addition, a space clipping operation is proposed based on the attention mechanism for constructing a new spatial search area. The search time can be further reduced by the space clipping operation under the path planning result within acceptable changes. Moreover, the entire optimization process and results can be feeded back to the knowledge database by the human-computer hybrid augmented intelligence framework to guide subsequent path planning issues. Finally, the simulation results confirm that a feasible and effective flight path can be quickly generated by the UAH path planning system based on human-computer hybrid augmented intelligence.

Hepatocellular carcinoma (HCC) is a common malignancy with poor prognosis and high mortality. To identify key genes associated with HCC and the underlying mechanisms, we performed weighted correlation network analysis (WGCNA) of potential key genes of HCC. We identified 17 key genes closely related to HCC by yellow module combined with PPI analysis. Verification of the role of these genes revealed that SPC25 knockdown results in a significant decrease in proliferation and metastasis of HCC cells and increased protein levels of components of the p53 pathway in vitro. In summary, we identified that SPC25 is a potential tumor‐promoting factor in HCC and may act via the p53 pathway., We report the differential expression of SPC25 in hepatocellular carcinoma (HCC) and its correlation with prognosis for the first time. Biological experiments verified that knockdown of SPC25 could significantly inhibit the proliferation and metastasis of HCC cells. In summary, this study shows that SPC25 plays an important role in HCC and is a potential prognostic biomarker and therapeutic target in HCC patients.

Graphical abstract, Cancer staging provides a common language that is used to describe the severity of an individual's cancer, which plays a critical role in optimizing cancer treatment. Recursive partitioning analysis (RPA) is the most widely accepted method for cancer staging. Despite its widespread use, to date, only limited tools have been developed to implement the RPA algorithm for cancer staging. Moreover, most of the available tools can be accessed only from command lines and also lack visualization, making them difficult for clinical investigators without programing skills to use. Therefore, we developed a web server called autoRPA that is dedicated to supporting the construction of prognostic staging models and performance comparisons among different staging models. Based on the RPA algorithm and log-rank test statistics, autoRPA can establish a decision-making tree from survival data and provide clinicians an intuitive method to further prune the decision tree. Moreover, autoRPA can evaluate the contribution of each submitted covariate that is involved in the grouping process and help identify factors that significantly contribute to cancer staging. Four indicators, including hazard consistency, hazard discrimination, percentage of variation explained, and sample size balance, are introduced to validate the performance of the designed staging models. In addition, autoRPA can also be used to compare the performance of different prognostic staging models using a standard bootstrap evaluation method. The web server of autoRPA is freely available at http://rpa.renlab.org.

Background The aim of this study is to investigate three-dimensional quantitative analysis of buccal augmented tissue alterations after surgery using a modified coronally advanced tunnel (MCAT) technique combined with a de-epithelialized gingival graft (DGG) within 1 year post-op, based on intraoral scanning. Methods 25 Cairo class I gingival recession defects were treated using an MCAT technique with DGG. Digital impressions were taken using an intraoral scanner at baseline, 2 weeks, 6 weeks, 3 months, and 1 year after the surgery. Three-dimensional quantitative measurements within 1 year were analyzed for buccal augmented tissue after surgery, including postoperative gingival height gain (GHG), area gain (GAG), volume gain (GVG) and mean thickness (GMT) of region of interest, as well as the tissue thickness change at 1, 2, and 3 mm (TTC1, TTC2, and TTC3) apical to the cemento-enamel junction. Results Postoperative GHG, GAG, GVG, and GMT were distinctly encountered at 2 weeks post-op, then gradually decreased. At 1 year, GHG, GAG, GVG, and GMT were 2.211 ± 0.717 mm, 7.614 ± 2.511 mm2, 7.690 ± 4.335 mm3 and 0.965 ± 0.372 mm, respectively. Significant decreases were recorded between 6 weeks and 1 year in terms of GHG, GAG, and GVG. The GMT was sustained after 6 weeks with an increase of nearly 1 mm at 1 year. TTC1 and TTC2 yielded thicker tissue change than TTC3. Conclusions Three-dimensional quantitative measurements taken via intraoral scanning showed that buccal augmented tissue acquired via MCAT with DGG tends to be stable after 3 months post-op. Digital measurement can be applied in periodontal plastic surgery as a clinically feasible and non-invasive evaluation method for achieving volumetric outcomes. Trial registration This study was retrospectively registered in the Chinese Clinical Trial Registry: ChiCTR1900026768. Date of registration: 21/10/2019.

Background (−)-Epigallocatechin Gallate (EGCG) as green tea catechins possessed antibacterial and anti-inflammatory effects on periodontal disease. This study was designed to evaluate the clinical and microbiological efficacy of scaling and root planing (SRP) using EGCG aqueous solution as coolants through a new-type ultrasonic scaler tip on chronic periodontitis. Methods This split-mouth, randomized clinical trial included 20 patients (2 drop-outs) with chronic periodontitis and the maxillary contra-lateral sides were allocated into test and control groups randomly. Through the new-type scaler tip, 762 sites with probing depth (PD) ≥ 4 mm were treated by SRP using EGCG solution or distilled water as coolants respectively. Clinical parameters and red complex pathogens in subgingival microbiome were evaluated at baseline, 3 and 6 months after treatments. Results During 6 months, the SRP plus EGCG medication contributed to additional PD reduction as 0.33 mm and gain of clinical attachment level as 0.3 mm compared with SRP alone, and approximate 8% more sites obtained PD reduction ≥ 2 mm (p Tannerella forsythia was significantly lower in the combined treatment group (p Conclusion The purified EGCG showed the potential to improve the outcome of periodontal non-surgical treatment and the new-type scaler tip provided an alternative vehicle for subgingival medication. Trial registration The trial was registered in Chinese Clinical Trial Registry on 15 February 2020 (No.: ChiCTR2000029831, retrospectively registered). http://www.chictr.org.cn/showprojen.aspx?proj=49441.

Background Interferon-alpha (IFNα) is a first-line treatment option for chronic hepatitis B virus (HBV) infection, but the severe systemic side-effects limited its clinical application. Interferon-lambda (IFNλ) with comparable antiviral activity and less toxic side-effects is thought to be a good alternative interferon to IFNα. Additionally, the gene vector mediated sustainably expression of therapeutic product in the target cells/tissue may overcome the shortcomings resulted from the short half-life of IFNs. Results We constructed a liver-specific IFNλ3-expressing minicircle (MC) vector under the control of a hepatocyte-specific ApoE promoter (MC.IFNλ3) and investigated its anti-HBV activity in a HBV-expressing hepatocyte-derived cell model (HepG2.2.15). As expected, the MC.IFNλ3 vector capable of expressing IFNλ3 in the recipient hepatocytes has demonstrated robust anti-HBV activity, in terms of suppressing viral antigen expression and viral DNA replication, via activation the interferon-stimulated gene (ISG) expression in HepG2.2.15 cells. Conclusions Given the MC vector can be easily delivered into liver, the liver-targeted IFN gene-transfer (MC.IFNλ3), instead of systemic administrating IFN repeatedly, provides a promising concept for the treatment of chronic HBV infection.

Background Although a series of reports on corneal fungal infection have been published, studies on pathogenic mechanisms and inflammation-associated cytokines remain limited. In this study, aqueous humor samples from fungal keratitis patients were collected to examine cytokine patterns and cellular profile for the pathogenesis of fungal keratitis. Methods The aqueous humor samples were collected from ten patients with advanced stage fungal keratitis. Eight aqueous humor samples from patients with keratoconus or corneal dystrophy were taken as control. Approximately 100 μl to 300 μl of aqueous humor in each case were obtained for examination. The aqueous humor samples were centrifuged and the cells were stained and examined under optical microscope. Bacterial and fungal cultures were performed on the aqueous humor and corneal buttons of all patients. Cytokines related to inflammation including IL-1β, IL-6, IL-8, IL-10, TNF-α, and IFN-γ were examined using multiplex bead-based Luminex liquid protein array systems. Results Fungus infection was confirmed in these ten patients by smear stains and/or fungal cultures. Bacterial and fungal cultures revealed negative results in all aqueous humor specimens. Polymorphonuclear leukocytes were the predominant infiltrating cells in the aqueous humor of fungal keratitis. At the advanced stages of fungal keratitis, the levels of IL-1β, IL-6, IL-8, and IFN-γ in the aqueous humor were significantly increased when compared with control (p

Background and Objective. The direct-acting antiviral agents (DAAs) antiviral therapy has drastically improved the prognosis of hepatitis C virus (HCV) patients. However, the viral drug resistance-associated variants (RAVs) can limit the efficacy of DAAs. For the HCV-6a is not the predominant prevalent genotype; the data on the prevalence of naturally occurring RAVs in it is scarce. Our study aims to assess the prevalence of RAVs in treatment-naive HCV-6a patients. Methods. Nested PCR assays were performed on 95 HCV-6a patients to amplify HCV viral regions of NS3, NS5A, and NS5B. Results. In NS3/4A region, we detected Q80K in 95.5% isolates (84/88) and D168E in 2.3% isolates (2/88). In NS5A region, we detected Q30R in 93.2% isolates (82/88), L31M in 4.6% isolates (4/88), and H58P in 6.8% isolates (6/88). In NS5B region, we detected A15G in 2.3% isolates (2/88), S96T in 1.1% isolates (1/88), and S282T in 20.7% isolates (17/88) and we detected I482L in 100% isolates (4/4), V494A in 50% isolates (2/4), and V499A in 100% isolates (4/4). Conclusions. RAVs to DAAs preexist in treatment-naive HCV-6a patients. Further studies should address the issue of the impact of RAVs in response to DAA therapies for HCV-6a patients.

The interest focusing on Eriobotrya japonica leaf triterpene acid (ELTA) has increased recently because of its beneficial effects on health. However, there has been a lack of experimental data on its toxicity. The present study therefore was conducted to evaluate its toxicity in ICR mice. The results showed that ELTA produced neither mortality nor toxicity of the main organs in ICR male and female mice in both acute (0.30, 0.65, 1.39, and 3.00 g·kg−1 body weight) and subacute (150, 300, and 600 mg·kg−1 BW) 28-day toxicity studies. Because of lacking apparently adverse effects found in the hematology, clinical biochemistry, and histopathology evaluation, results of the present study together with the beneficial effects make ELTA as a promising dietary supplement and indicated that ELTA administered orally might have a large safety margin for human.

BackgroundThrough the comparison of three-dimensional CT reconstruction between the supine position and the prone position, the relative position of thoracolumbar great vessels and vertebral body was studied, and the shortest safe distance between them was measured to improve the safety of bicortical pedicle screw insertion and reduce the risk of vascular injury.MethodsForty adults were selected to participate the research. Three-dimensional reconstruction of thoracolumbar (T9-L3) CT was performed in the prone position and the supine position. The relative distance between the Aorta/Inferior Vena Cava (IVC) and vertebral body was obtained as AVD/VVD respectively. The relative angle of the Aorta/ IVC and the vertebral body was calculated as ∠AOY/∠VOY. Self-controlled experiments were carried out in the prone and the supine positions, and the data obtained were analyzed using SPSS 22.0 statistical software.ResultsThe AVD of the prone position and the supine position was the shortest at T12 (3.18 ± 0.68 mm), but the difference was not statistically significant. The aorta of the T9-L3 segment was shifted from the anterolateral to the anteromedial. The ∠AOY of the other groups differed significantly between the prone and supine positions in all vertebrae except T12 and L1 (P With regard to VVD/∠VOY, there was no significant difference between the prone and supine positions (P ≥ 0.05), and the minimum VVD of L3 segment is greater than 5.4 mm. The IVC has no obvious mobility and is fixed in the range of 20 ° ~ 30 ° near the midline.ConclusionWhen using bicortical anchoring of pedicle screws, it is safe to ensure that the protruding tips of the screw is less than 3 mm. Due to the mobility of the aorta in different postures and individual differences in anatomy, the prone position CT can help doctors to make better preoperative plans and decisions.

Rationale: Femoral intertrochanteric fracture happens easily in the elderly, especially those with osteoporosis. As a standard intramedullary fixation implant, Proximal Femoral Nail Anti-rotation (PFNA) is applied to various types of stable or unstable intertrochanteric fractures of femur. Due to blade-related factors, such as cutting-through into the hip joint, cutting out or back out, there are endless postoperative failure cases of internal fixation, but reports about perioperative failure that the helical blade cannot be tightened are lacking. In this case, we firstly report an intraoperative blade compression failure and an effective technique to help surgeons out of the dilemma by using cannulated hexagonal screwdriver which was already included in the orthopedic instrument box. Patient concerns: An 81-year-old lady suffered left hip pain, swelling and limitation of activity, after slipping and falling when she was in the toilet. Diagnoses: X-ray and computed tomography (CT)-scan showed comminuted left intratrochanteric fracture with a Jensen classification of IIa type. Interventions: The patient was treated by closed reduction and internal fixation with PFNA. We suffered an unprecedented problem that helical blade cannot be tightened by the blade impactor as usual. For fear of helical blade disintegration during removal and a significant decrease in pullout strength after reinsertions, we eliminated the dilemma by using a cannulated hexagonal screwdriver to tighten the “problem” helical blade. Outcomes: The “problem” helical blade was finally locked by using the SW4.0 mm cannulated hexagonal screwdriver which was already included in the instrument box. The operation ended successfully after completing subsequent steps. Lessons: The cannulated hexagonal screwdriver is an effective instrument that can help surgeons out of the dilemma when the blade impactor fails to tighten the helical blade in PFNA implantation.

Background: Hoffa fractures are coronal-plane fractures of the femoral condyle, which are rarer than sagittal-plane condylar fractures. This study aimed to systematically review the clinical knowledge base of Hoffa fractures to facilitate the diagnosis and management of such injuries. Methods: We searched Medline, Embase, Cochrane Library, Google Scholar, China National Knowledge Infrastructure, and China Biology Medicine disc, using the terms “Hoffa fracture” and “coronal fracture of femoral condyle.” Results: One hundred five articles on Hoffa fractures were reviewed, and the clinical knowledge base was summarized. High-energy trauma is a common cause of a Hoffa fracture, although low-energy trauma and iatrogenic injury can also lead to these fractures. Commonly used classifications include the Letenneur classification, a computed tomography (CT) classification, the AO classification, and modified AO classification. Radiography can reveal fracture lines. If radiographic findings are negative in questionable cases, CT and magnetic resonance imaging (MRI) should be performed. Nondisplaced fractures can be managed conservatively; however, they involve a high risk of redisplacement. Open reduction and internal fixation are preferred. For young patients with good compliance, simple medial or lateral condylar fractures can be treated via a medial or lateral parapatellar approach. After fracture exposure, headless compression screws can be inserted perpendicularly to the fracture line from posterior to anterior. For bicondylar fractures, a median parapatellar incision can be used. For complex fractures in patients with osteoporosis or a high body mass index, cannulated screws with antigliding plate fixation should be used. Conclusion: Here, we summarized the injury mechanism, diagnosis, classification, and treatment options of Hoffa fractures.

This paper develops two-stage inverse data envelopment analysis models with undesirable outputs to formulate resource plans for 16 Chinese listed commercial banks whose outputs are increased and overall efficiency is kept unchanged in the short term. We use these models to meet three different output targets, namely, increasing both the desirable and undesirable outputs by the same percentage, increasing these outputs by different percentages, and increasing only the desirable outputs while keeping the undesirable outputs unchanged. We find that operation cost and interest expense are more flexible than labor in the adjustment process and that deposits have no obvious law of change. The findings of this work provide some suggestions for bank managers.

Rationale: Traumatic hip dislocation is a common joint dislocation. Delayed reduction has been shown to increase the risk of avascular necrosis of the femoral head. Most of the traditional methods must be performed under general anesthesia or spinal anesthesia to relax hip muscles. Anesthesia will prolong the interval between the injury and the reduction. Patient concerns: 16 patients presented with hip pain and a leg shortened, flexed, internally rotated and adducted. Diagnoses: X-ray and CT-scan showed acute closed posterior dislocation of hip. Interventions: Closed reduction of the traumatic posterior-dislocation of hip joint using a novel sitting technique. Outcomes: Among these 16 patients, 15 hips were successfully reduced using the Sitting Technique (table 1), indicating the success rate was 93.8%(15/16). A total of 12 patients were followed up, with a mean period of 23.5 months (range, 6–72 months). Among these 12 patients, 10 patients (83.3%) had excellent grade, 2 patients (16.7%) had good grade. Lessons: Sitting technique for treatment of traumatic posterior dislocation of hip joint does not need anaesthesia, which it shortens the interval between the injury and the reduction and saves valuable time for 6 hours of joint reduction. On the other hand, this method does no harm to the physicians’ low back.

This broad study identifies landfill emission reduction cobenefits in China, as well as implications for policy and practice., China’s CH4 emissions from 1955 existing (old) and 495 planned (new) landfills are estimated and projected by adopting a bottom-up method, targeting a 2012 baseline year and a 2030 projected target year. Nine key CH4 mitigation measures are proposed for the CH4 mitigation assessment from landfills. Approximately 0.66 million metric tons (Mt) of CH4 and 1.14 Mt of CH4 will be released, respectively, from new and existing landfills under a 2030 business-as-usual (BAU) scenario, which is 23.5% lower than a U.S. Environmental Protection Agency estimation. It is estimated that 0.60 and 0.97 Mt of CH4 can be reduced under new policies (NP) and low-carbon (LC) policy scenarios, respectively. The combined biocover and landfill gas collection and flaring system is the most promising mitigation measure, while mechanical biological treatment and mineral landfill also contribute substantially to CH4 reduction. The odor-affected population under NP and LC scenarios will decrease by 39.5 and 64.2%, respectively, when compared to the 2030 BAU scenario. The LC scenario is a recommended policy for meeting China’s nationally determined contribution targets and reducing the not-in-my-backyard impact due to this policy’s significant reduction of CH4 emissions.

To observe the clinical outcome of the sinus tarsi approach in the operative treatment of intra-articular calcaneal fractures. Forty-nine intra-articular calcaneal fractures in 45 patients were managed surgically with sinus tarsi approach. The anatomical plate and compression bolts were applied in 14 feet. The anatomical plate and screws were applied in 35 feet. Maryland foot score system was used to evaluate the function of the hindfoot at the followup. The reduction of the posterior facet was graded as nearly anatomical (less than 2 mm articular displacement) in 46 feet (93.9%). The width, height, and Böhler angle were significantly improved in all patients (P

Supplemental Digital Content is available in the text, The direct-acting antiviral agents (DAAs) have drastically improved the prognosis of hepatitis C virus (HCV) patients. However, the resistance-associated variants (RAVs) to DAAs may hamper treatment. There was a lack of data on the prevalence of pre-exist RAVs in Chinese HCV-infected patients. We performed nested PCR assays on 74 HCV genotype 1b-infected patients to amplify HCV viral regions of NS3, NS5A, and NS5B to investigate the prevalence of RAVs to DAAs in treatment-naive HCV genotype1b-infected patients in China. The mutations A156S, T54S, and D168Y of the NS3/4A region were found in 18.33% (11/60), 6.67% (4/60), and 1.67% (1/60) of the successfully amplified cases. Mutations Q30R, L31M, and H58P of the NS5A region were confirmed in 57.63% (34/59), 1.69%(1/59), and 86.44% (51/59) of the cases. Mutations C316N, S365A, M414L, M423I, Y448H, I482T, I482 V, V494L, P495S, and V499A of the NS5B region were detected in 100% (60/60), 3.33% (2/60), 5.88% (3/51), 1.96% (1/51), 1.96% (1/51), 5.88% (3/51), 1.96% (1/51), 3.92% (2/51), 5.88% (3/51), and 15.69% (8/51) of cases, respectively. Naturally occurring RAVs to DAAs pre-exist in treatment-naive Chinese HCV genotype 1b-infected patients and the characteristic is different from that in Europe and the United States. Clinicians should consider RAVs upon the introduction of DAA-based antiviral therapy.

The genes of major histocompatibility complex (MHC) are important to vertebrate immune system. In this study, two new MHC class I genes, designated as Egeu-UAA and Egeu-UBA, were discovered in the vulnerable Chinese egret (Egretta eulophotes). Using a full length DNA and cDNA produced by PCR and RACE methods, these two MHC class I loci were characterized in the genome of the Chinese egret and were also found to be expressed in liver and blood. Both new genes showed the expected eight exons and were similar to two copies of the minimal essential MHC complex of chicken. In genetic diversity, 14 alleles (8 for UAA and 6 for UBA) in the MHC class I gene exon 3 were found in 60 individuals using locus-specific primers and showed little polymorphism. Only three potential amino acid residues were detected under positive selection in potential peptide-binding regions (PBRs) by Bayesian analysis. These new results provide the fundamental basis for further studies to elucidate the molecular mechanisms and significance of MHC molecular adaptation in vulnerable Chinese egret and other ardeids, finding that have not been previously reported.

Few studies have conducted follow-up investigations of the clinical course in HCV-related cirrhotic patients who achieved a sustained virological response (SVR) with pegylated interferon plus ribavirin treatment (PegIFN + RBV). We investigated the clinical course and laboratory data in a prospective cohort study enrolling HCV-related cirrhotic patients who received PegIFN + RBV between August 2008 and July 2013 in China. Complete blood counts, liver function tests, and HCV-RNA were serially examined. Liver-related complications were recorded. To detect hepatocellular carcinoma (HCC), alpha-fetoprotein assays, and ultrasound scans were repeated at 6-month intervals. Twenty-five patients were enrolled, including 8 patients with decompensation events before treatment. Eighteen patients achieved SVR with a mean follow-up period of 25.78 months. During the follow-up period, only one patient exhibited HCV-RNA positivity and no decompensation events were detected, but 4 patients developed HCC after SVR. APRI decreased more in patients with SVR than in patients with non-SVR (median, −1.33 versus 0.86,P<0.001). The albumin levels and platelet counts significantly increased during the follow-up period after SVR (44.27±4.09versus42.63±4.37,P=0.037and173.89±87.36versus160.11±77.97,P=0.047). These data indicated that HCV-related cirrhotic patients with SVR after PegIFN + RBV may have a favorable clinical course and improvements in laboratory data. Moreover, HCC should be monitored.

Background IgA nephropathy (IgAN) is the most common form of primary glomerulonephritis worldwide. In recent years, consistent efforts have been made to develop new non-invasive biomarkers for IgAN progression. In our previous in vitro study we found mesangial derived CXCL1 as a contributor for kidney injury, and observed higher urinary CXCL1 levels in patients with IgAN. It implied that the urinary CXCL1 might be a potential biomarker. Methods In the present study, we enrolled 425 IgAN patients with follow-up data and detected their urinary CXCL1 levels at the time of renal biopsy, to explore the predictive value of urinary CXCL1 in IgAN progression. Urinary CXCL1 levels were measured using enzyme-linked immunosorbent assay. Results Urinary CXCL1 levels were associated with presently well established predictors of IgAN progression, including SBP (r = 0.138, p = 0.004), DBP (r = 0.114, p = 0.019), proteinuria (r = 0.155, p = 0.001), eGFR (r = -0.259, p

The inhibition of NF-κB by genetic deletion or pharmacological inhibition of IKK2 significantly reduces laser-induced choroid neovascularization (CNV). To achieve a sustained and controlled intraocular release of a selective and potent IKK2 inhibitor, 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1) (MW: 279.29), we developed a biodegradable poly-lactide-co-glycolide (PLGA) polymer-delivery system to further investigate the anti-neovascularization effects of IKK2 inhibition and in vivo biosafety using laser-induced CNV mouse model. The solvent-evaporation method produced spherical TPCA-1-loaded PLGA microparticles characterized with a mean diameter of 2.4 ¼m and loading efficiency of 80%. Retrobulbar administration of the TPCA-1-loaded PLGA microparticles maintained a sustained drug level in the retina during the study period. No detectable TPCA-1 level was observed in the untreated contralateral eye. The anti-CNV effect of retrobulbarly administrated TPCA-1-loaded PLGA microparticles was assessed by retinal fluorescein leakage and isolectin staining methods, showing significantly reduced CNV development on day 7 after laser injury. Macrophage infiltration into the laser lesion was attenuated as assayed by choroid/RPE flat-mount staining with anti-F4/80 antibody. Consistently, laser induced expressions of Vegfa and Ccl2 were inhibited by the TPCA-1-loaded PLGA treatment. This TPCA-1 delivery system did not cause any noticeable cellular or functional toxicity to the treated eyes as evaluated by histology and optokinetic reflex (OKR) tests; and no systemic toxicity was observed. We conclude that retrobulbar injection of the small-molecule IKK2 inhibitor TPCA-1, delivered by biodegradable PLGA microparticles, can achieve a sustained and controllable drug release into choroid/retina and attenuate laser-induced CNV development without causing apparent systemic toxicity. Our results suggest a potential clinical application of TPCA-1 delivered by microparticles in treatment of CNV in the patients with age-related macular degeneration and other retinal neovascularization diseases.

Mertk belongs to the Tyro3, Axl and Mertk (TAM) family of receptor tyrosine kinases, and plays a pivotal role in regulation of cytoskeletal rearrangement during phagocytosis. Phagocytosis by either professional or non-professional phagocytes is impaired in the Mertk deficient individual. In the present study, we further investigated the effects of Mertk mutation on peritoneal macrophage morphology, attachment, spreading and movement. Mertk-mutated macrophages exhibited decreased attachment, weak spreading, loss of spindle-like body shape and lack of clear leading and trailing edges within the first few hours of culture, as observed by environmental scanning electron microscopy. Time-lapse video photography recording showed that macrophage without Mertk conducted mainly random movement with oscillating swing around the cell body, and lost the directional migration action seen on the WT cells. Western blotting showed a decreased phosphorylation of focal adhesion kinase (FAK). Immunocytochemistry revealed that actin filaments and dynamic protein myosin II failed to concentrate in the leading edge of migrating cells. Microtubules were localized mainly in one side of mutant cell body, with no clear MTOC and associated radially-distributed microtubule bundles, which were clearly evident in the WT cells. Our results suggest that Mertk deficiency affects not only phagocytosis but also cell shape and migration, likely through a common regulatory mechanism on cytoskeletons.

Background The animal mitochondrial genome is generally considered to be under selection for both compactness and gene order conservation. As more mitochondrial genomes are sequenced, mitochondrial duplications and gene rearrangements have been frequently identified among diverse animal groups. Although several mechanisms of gene rearrangement have been proposed thus far, more observational evidence from major taxa is needed to validate specific mechanisms. In the current study, the complete mitochondrial DNA of sixteen bird species from the family Ardeidae was sequenced and the evolution of mitochondrial gene rearrangements was investigated. The mitochondrial genomes were then used to review the phylogenies of these ardeid birds. Results The complete mitochondrial genome sequences of the sixteen ardeid birds exhibited four distinct mitochondrial gene orders in which two of them, named as “duplicate tRNAGlu–CR” and “duplicate tRNAThr–tRNAPro and CR”, were newly discovered. These gene rearrangements arose from an evolutionary process consistent with the tandem duplication - random loss model (TDRL). Additionally, duplications in these gene orders were near identical in nucleotide sequences within each individual, suggesting that they evolved in concert. Phylogenetic analyses of the sixteen ardeid species supported the idea that Ardea ibis, Ardea modesta and Ardea intermedia should be classified as genus Ardea, and Ixobrychus flavicollis as genus Ixobrychus, and indicated that within the subfamily Ardeinae, Nycticorax nycticorax is closely related to genus Egretta and that Ardeola bacchus and Butorides striatus are closely related to the genus Ardea. Conclusions The duplicate tRNAThr–CR gene order is found in most ardeid lineages, suggesting this gene order is the ancestral pattern within these birds and persisted in most lineages via concerted evolution. In two independent lineages, when the concerted evolution stopped in some subsections due to the accumulation of numerous substitutions and deletions, the duplicate tRNAThr–CR gene order was transformed into three other gene orders. The phylogenetic trees produced from concatenated rRNA and protein coding genes have high support values in most nodes, indicating that the mitochondrial genome sequences are promising markers for resolving the phylogenetic issues of ardeid birds when more taxa are added. Electronic supplementary material The online version of this article (doi:10.1186/1471-2164-15-573) contains supplementary material, which is available to authorized users.

The major histocompatibility complex (MHC) is an excellent molecular marker for the studies of evolutionary ecology and conservation genetics because it is a family of highly polymorphic genes that play a key role in vertebrate immune response. In this study, the functional genes of MHC Class II B (DAB) were isolated for the first time in a vulnerable species, the Chinese egret ( Egretta eulophotes ). Using a full length DNA and cDNA produced by PCR and RACE methods, four potential MHC DAB loci were characterized in the genome of this egret and all four were expressed in liver and blood. At least four copies of the MHC gene complex were similar to two copies of the minimal essential MHC complex of chicken, but are less complex than the multiple copies expressed in passerine species. In MHC polymorphism, 19 alleles of exon 2 were isolated from 48 individuals using PCR. No stop codons or frameshift mutations were found in any of the coding regions. The signatures of positive selection detected in potential peptide-binding regions by Bayesian analysis, suggesting that all of these genes were functional. These data will provide the fundamental basis for further studies to elucidate the mechanisms and significance of MHC molecular adaptation in vulnerable Chinese egret and other ardeids.

The Tyro3, Axl and Mertk (TAM) triply knockout (TKO) mice exhibit systemic autoimmune diseases, with characteristics of increased proinflammatory cytokine production, autoantibody deposition and autoreactive lymphocyte infiltration into a variety of tissues. Here we show that TKO mice produce high level of serum TNF-α and specific autoantibodies deposited onto brain blood vessels. The brain-blood barrier (BBB) in mutant brains exhibited increased permeability for Evans blue and fluorescent-dextran, suggesting a breakdown of the BBB in the mutant brains. Impaired BBB integrity facilitated autoreactive T cells infiltrating into all regions of the mutant brains. Brain autoimmune disorder caused accumulation of the ubiquitin-reactive aggregates in the mutant hippocampus, and early formation of autofluorescent lipofuscins in the neurons throughout the entire brains. Chronic neuroinflammation caused damage of the hippocampal mossy fibers and neuronal apoptotic death. This study shows that chronic systemic inflammation and autoimmune disorders in the TKO mice cause neuronal damage and death.

Purpose The Notch signaling pathway plays crucial roles in regulation of cell proliferation, differentiation and cell fate decision in multiple tissues and cell types. This study was designed to test the effects of enhanced Notch activity on corneal epithelium homeostasis and wound healing using the transgenic mice that overexpressed an activated Notch1 (NICD) in cornea epithelium. Methods The studies were performed on R26fN1-ICD transgenic mice that carry a NICD cDNA (cDNA) whose expression is prevented by a “Lox-STOP-Lox” cassette. When this transgenic mouse is bred to a mouse strain carrying a Cre recombinase expression cassette driven by a tissue-specific keratin 14 (K14) promoter, the floxed “STOP” cassette is excised and NICD is expressed in the cornea epithelium. The expression level of NICD and its downstream target genes, hairy and enhancer of split 1 (Hes1) and hairy/enhancer-of-split related with YRPW motif 1 (Hey1), in the transgenic corneal epithelium was examined by quantitative PCR (qPCR). The phenotypes and morphology of the transgenic corneal epithelium were compared with that of wild type (WT) controls. The proliferation rate of the epithelial cells was assessed by 5-bromo-2'-deoxyuridine (BrdU) incorporation and the differentiation statues were examined by K14, tumor protein p63 (p63), K12, and zona occludens 1 (ZO-1) immunoreactivity at either normal developmental condition or after corneal epithelial debridement. The corneal epithelial response to wound healing was studied by fluorescent staining and Richardson’s staining macroscopically and by H&E staining at microscope level at 0, 6, 12, 18, and 24 h post injury. Results Although overexpression of NICD in cornea epithelium led to upregulation of its downstream targets, i.e., Hes1 and Hey1, this did not alter corneal epithelial cell proliferation and differentiation. However, wound healing induced Notch activity and overexpression of NICD promoted corneal epithelial wound healing, which was in agreement with more rapid early proliferation response in NICD transgenic mice than in the wild type control mice. Conclusions These findings further demonstrate the functional role of Notch signaling in corneal epithelium wound healing response.

Diabetic nephropathy is a complex and poorly understood disease process, and our current treatment options are limited. It remains critical, then, to identify novel therapeutic targets. Recently, a developmental protein and one of the bone morphogenetic protein antagonists, Gremlin, has emerged as a novel modulator of diabetic nephropathy. The high expression and strong co-localization with transforming growth factor-beta1 in diabetic kidneys suggests a role for Gremlin in the pathogenesis of diabetic nephropathy. We have constructed a gremlin siRNA plasmid and have examined the effect of Gremlin inhibition on the progression of diabetic nephropathy in a mouse model. CD-1 mice underwent uninephrectomy and STZ treatment prior to receiving weekly injections of the plasmid. Inhibition of Gremlin alleviated proteinuria and renal collagen IV accumulation 12 weeks after the STZ injection and inhibited renal cell proliferation and apoptosis. In vitro experiments, using mouse mesangial cells, revealed that the transfect ion of gremlin siRNA plasmid reversed high glucose induced abnormalities, such as increased cell proliferation and apoptosis and increased collagen IV production. The decreased matrix metalloprotease level was partially normalized by transfection with gremlin siRNA plasmid. Additionally, we observed recovery of bone morphogenetic protein-7 signaling activity, evidenced by increases in phosphorylated Smad 5 protein levels. We conclude that inhibition of Gremlin exerts beneficial effects on the diabetic kidney mainly through maintenance of BMP-7 activity and that Gremlin may serve as a novel therapeutic target in the management of diabetic nephropathy.

Objectives The optimal treatment of hepatitis C virus (HCV) genotype 6 is unclear owing to its limited geographic distribution. Because of a high predictive value of rapid virological response (RVR) for sustained virological response (SVR), we conducted an open-label randomized controlled trial to compare 24- and 48-week peginterferon/ribavirin combination therapy for patients with HCV genotype 6 in Southern China who achieved an RVR. Methods and Findings Treatment-naive, non-cirrhotic patients with chronic hepatitis C genotype 6 were treated with pegylated interferon α-2a (180 μg/week) and ribavirin (800–1,200 mg, according to weight) for 4 weeks. Patients who achieved an RVR, which was defined as HCV RNA negativity at week 4 (

Tyro3, Axl and Mertk (TAM) receptor tyrosine kinases play multiple functional roles by either providing intrinsic trophic support for cell growth or regulating the expression of target genes that are important in the homeostatic regulation of immune responses. TAM receptors have been shown to regulate adult hippocampal neurogenesis by negatively regulation of glial cell activation in central nervous system (CNS). In the present study, we further demonstrated that all three TAM receptors were expressed by cultured primary neural stem cells (NSCs) and played a direct growth trophic role in NSCs proliferation, neuronal differentiation and survival. The cultured primary NSCs lacking TAM receptors exhibited slower growth, reduced proliferation and increased apoptosis as shown by decreased BrdU incorporation and increased TUNEL labeling, than those from the WT NSCs. In addition, the neuronal differentiation and maturation of the mutant NSCs were impeded, as characterized by less neuronal differentiation (β-tubulin III+) and neurite outgrowth than their WT counterparts. To elucidate the underlying mechanism that the TAM receptors play on the differentiating NSCs, we examined the expression profile of neurotrophins and their receptors by real-time qPCR on the total RNAs from hippocampus and primary NSCs; and found that the TKO NSC showed a significant reduction in the expression of both nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), but accompanied by compensational increases in the expression of the TrkA, TrkB, TrkC and p75 receptors. These results suggest that TAM receptors support NSCs survival, proliferation and differentiation by regulating expression of neurotrophins, especially the NGF.

Choroidal neovascularization (CNV) is aberrant angiogenesis associated with exudative age-related macular degeneration (AMD), a leading cause of blindness in the elderly. Inflammation has been suggested as a risk factor for AMD. The IKK2/NF-κB pathway plays a key role in the inflammatory response through regulation of the transcription of cytokines, chemokines, growth factors and angiogenic factors. We investigated the functional role of IKK2 in development of the laser-induced CNV using either Ikk2 conditional knockout mice or an IKK2 inhibitor. The retinal neuronal tissue and RPE deletion of IKK2 was generated by breeding Ikk2(-/flox) mice with Nestin-Cre mice. Deletion of Ikk2 in the retina caused no obvious defect in retinal development or function, but resulted in a significant reduction in laser-induced CNV. In addition, intravitreal or retrobulbar injection of an IKK2 specific chemical inhibitor, TPCA-1, also showed similar inhibition of CNV. Furthermore, in vitro inhibition of IKK2 in ARPE-19 cells significantly reduced heat shock-induced expression of NFKBIA, IL1B, CCL2, VEGFA, PDGFA, HIF1A, and MMP-2, suggesting that IKK2 may regulate multiple molecular pathways involved in laser-induced CNV. The in vivo laser-induced expression of VEGFA, and HIF1A in RPE and choroidal tissue was also blocked by TPCA-1 treatment. Thus, IKK2/NF-κB signaling appears responsible for production of pro-inflammatory and pro-angiogenic factors in laser-induced CNV, suggesting that this intracellular pathway may serve as an important therapeutic target for aberrant angiogenesis in exudative AMD.

Podocyte loss has been reported to relate to disease severity and progression in IgA nephropathy (IgAN). However, the underlying mechanism for its role in IgAN remain unclear. Recent evidence has shown that IgA1 complexes from patients with IgAN could activate mesangial cells to induce soluble mediator excretion, and further injure podocytes through mesangial-podocytic cross-talk. In the present study, we explored the underlying mechanism of mesangial cell-induced podocyte loss in IgAN. We found that IgA1 complexes from IgAN patients significantly up-regulated the expression of CXCL1 and TGF-β1 in mesangial cells compared with healthy controls. Significantly higher urinary levels of CXCL1 and TGF-β1 were also observed in patients with IgAN compared to healthy controls. Moreover, IgAN patients with higher urinary CXCL1 and TGF-β1 presented with severe clinical and pathological manifestations, including higher 24-hour urine protein excretion, lower eGFR and higher cresentic glomeruli proportion. Further in vitro experiments showed that increased podocyte death and reduced podocyte adhesion were induced by mesangial cell conditional medium from IgAN (IgAN-HMCM), as well as rhCXCL1 together with rhTGF-β1. In addition, the over-expression of CXCR2, the receptor for CXCL1, by podocytes was induced by IgAN-HMCM and rhTGF-β1, but not by rhCXCL1. Furthermore, the effect of increased podocyte death and reduced podocyte adhesion induced by IgAN-HMCM and rhCXCL1 and rhTGF-β1 was rescued partially by a blocking antibody against CXCR2. Moreover, we observed the expression of CXCR2 in urine exfoliated podocytes in IgAN patients. Our present study implied that IgA1 complexes from IgAN patients could up-regulate the secretion of CXCL1 and TGF-β1 in mesangial cells. Additionally, the synergistic effect of CXCL1 and TGF-β1 further induced podocyte death and adhesion dysfunction in podocytes via CXCR2. This might be a potential mechanism for podocyte loss observed in IgAN.