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Background Gastric cancer (GC) is one of the most common tumors and the major cause of cancer-related mortality in the world. The purpose of this study is to identify new biomarker and reveal its potential molecular mechanism in GC. Methods The expression of CAP2 was observed by the bioinformatics analysis and western blot assays. The effects of CAP2 on cell proliferation and growth were tested by MTT assay, EdU assay, colony formation assay, and flow cytometric assay, respectively. ChIP and dual-luciferase assays were confirmed that SOX9 binding sites were putative regulatory elements in the transcriptional activation of CAP2. Furthermore, western blot and xenograft assays were applied to examine whether SOX9 was involved in the regulation of CAP2 expression. Results We reported that CAP2 is overexpressed in GC cells and tissues and related to a poorer prognosis for GC patients. Moreover, we found that knockdown of CAP2 suppressed the proliferation, growth, and cell cycle of GC cells. Besides, the transcription factor SOX9 participated in the CAP2-mediated proliferation of GC cells in vitro and in vivo. Conclusions Our results provide novel evidence that CAP2 plays an essential role in the genesis and development of GC, thus potentially highlighting this gene as a therapeutic target.

This study compares the longitudinal histological characteristics of proximal humeral implants with different spatial structures in rabbits. Thirty skeletally-mature male rabbits were divided into a trabecular structure group and regular hexahedron structure group according to the different spatial structures of a biological titanium alloy screw inserted into the greater tuberosity of the proximal humerus. Samples were collected 3, 6, and 12 weeks after the implantation surgery. Histological results showed that the amount of bone in-growth in the porous cavity of the screw implant increased over time. Quantitative analysis showed there was significantly more bone in-growth in the trabecular structure group than the classic structure group 3 weeks (25.4% ± 6.9% vs 19.6% ± 3.7%, P < 0.05) and 6 weeks (31.2% ± 1.7% vs 26.9% ± 5.3, P < 0.05) after the implantation surgery. No significant difference was detected between the two groups 12 weeks after the surgery (41.7% ± 2.5% vs 39% ± 4.1%, P > 0.05). Our data found that bone in-growth significantly differed among the three time points (P < 0.05) in both groups, but not between the implants with different spatial structures 12 weeks after the surgery.

In this study, we investigated whether the measurement of patellar tracking can be used as a diagnostic parameter of patellofemoral joint disease. Patellar tracking is defined as the movement of the patella in relation to the femorotibial joint within the full range of flexion and extension of the knee joint. The PubMed, EMBASE, Medline, PsychINFO, and AMED databases were used to find relevant articles. Analyzed were the patellar tracking coordinate system and the measurement objects, precision, methods used in those studies, as well as the results obtained. Origin points for coordinate systems varied across the studies. The research object and methods of patellar tracking varied in the studies. Most studies focused on a static description of the internal and external displacement and the internal and external inclination. The in vivo, noninvasive, and six degrees of freedom evaluation of patellar tracking reflect patellar motion more comprehensively, though each of these methods does so in different ways. Dynamic and quantitative evaluation of patellar tracking is still lacking in clinical work. Accurate and quantitative patellar tracking measurement could provide clinicians with a comprehensive evaluation of the stability of the knee joint.

The majority of circular RNAs (circRNAs) spliced from coding genes contain open reading frames (ORFs) and thus, have protein coding potential. However, it remains unknown what regulates the biogenesis of these ORF-containing circRNAs, whether they are actually translated into proteins and what functions they play in specific physiological contexts. Here, we report that a large number of circRNAs are synthesized with increasing abundance when late pachytene spermatocytes develop into round and then elongating spermatids during murine spermatogenesis. For a subset of circRNAs, the back splicing appears to occur mostly at m6A-enriched sites, which are usually located around the start and stop codons in linear mRNAs. Consequently, approximately a half of these male germ cell circRNAs contain large ORFs with m6A-modified start codons in their junctions, features that have been recently shown to be associated with protein-coding potential. Hundreds of peptides encoded by the junction sequences of these circRNAs were detected using liquid chromatography coupled with mass spectrometry, suggesting that these circRNAs can indeed be translated into proteins in both developing (spermatocytes and spermatids) and mature (spermatozoa) male germ cells. The present study discovered not only a novel role of m6A in the biogenesis of coding circRNAs, but also a potential mechanism to ensure stable and long-lasting protein production in the absence of linear mRNAs, i.e., through production of circRNAs containing large ORFs and m6A-modified start codons in junction sequences.

Chronic kidney disease (CKD) is associated with higher risk of cardiovascular disease-related ischemic events, which includes peripheral arterial disease (PAD). PAD is a strong predictor of future cardiovascular events, which can cause significant morbidity and mortality. Resistin has been found to be involved in pathological processes leading to CVD. Therefore, we aim to investigate whether resistin level is correlated with PAD in patients with non-dialysis CKD stage 3 to 5. There were 240 CKD patients enrolled in this study. Ankle-brachial index (ABI) values were measured using the automated oscillometric method. An ABI value < 0.9 defined the low ABI group. Serum levels of human resistin were determined using a commercially available enzyme immunoassay. Thirty CKD patients (12.5%) were included in the low ABI group. Patients in the low ABI group were older and had higher resistin levels as well as higher diabetes mellitus, hypertension and habit of smoking, and lower estimated glomerular filtration rate than patients in the normal ABI group. After the adjustment for factors that were significantly associated with PAD by multivariate logistic regression analysis, age and serum resistin level were independent predictors of PAD. A high serum resistin level is an independent predictor of PAD in non-dialysis CKD stage 3 to 5.

Background This study aimed to investigate the differences in the serum levels of glucose, lipid, and thyroid function markers between unipolar and bipolar depressed patients, as well as the effect of anhedonia and suicidal thoughts on the levels of these biochemical parameters. Methods A total of 287 unmedicated depressed patients from January 2016 to December 2017 were included in this study, including 92 bipolar depressions and 195 unipolar depressions. Anhedonia was determined using the item 32 of Symptom Checklist (SCL-90). Suicide ideation was assessed by item 15 of SCL-90. Results The bipolar group had significantly lower lipid levels (including triglycerides, cholesterol, low-density lipoprotein cholesterol [LDL], very low-density lipoprotein cholesterol [VLDL]) and insulin resistance index but higher levels of prolactin, low triiodothyronine (T3) and free T3 (FT3) as well as higher incidence of anhedonia as compared with the unipolar group. Depressed patients with anhedonia had significantly higher LDL level than those without anhedonia. Depressed patients with suicidal thoughts had cholesterol and high-density lipoprotein cholesterol (HDL) level. The above-mentioned differences were confirmed by logistic regression analysis. Receiver operating characteristic curve (ROC) analysis showed that the area under the ROC curve (AUC) ranged from 0.546 to 0.685. Conclusion Triglycerides, cholesterol, LDL, VLDL T3, FT3 levels were significantly different between unipolar and bipolar depressed patients, which might have the potential to be the markers for differential diagnosis. Patients with anhedonia had lower LDL level, while patients with suicidal thoughts had higher levels of cholesterol and HDL as compared with the corresponding control groups.

Artemisinin is an important frontline antimalarial. Fast, accurate detection of artemisinin in human serum is of importance in monitoring its clinical pharmaceutical effect. In this work, a strategy using microsensor array coupled with electrochemiluminescence (ECL) imaging technique was developed for detection of artemisinin. The microsensor array was constructed by integrating a patterned indium tin oxide glass plate with two perforated hydrophobic paper covers. By introducing the reactant of p-aminophenylboronic acid, luminol and artemisinin into the microsensor array, artemisinin would oxidize p-aminophenylboronic acid into p-aminophenol, a product which can efficiently inhibit the ECL of luminol. ECL signals decrease linearly with the increase of artemisinin. Based on the decreased ECL signal, artemisinin can be accurately detected. A good linearity (r = 0.994) was observed for artemisinin detection. The detection sensitivity is 0.48 μM for artemisinin. The detection selectivity and stability were also investigated. Results show that the present method shows a good selectivity and stability towards artemisinin detection. To evaluate the applicability of the present strategy for detecting artemisinin in real samples, the artemisinin content in human serum and Artemisia annua samples were analyzed. Results demonstrated that the present strategy shows excellent selectivity with high sensitivity towards artemisinin detection in real samples.

Cilia are cell-surface, microtubule-based organelles that project into extracellular space. Motile cilia are conserved throughout eukaryotes, and their beat induces the flow of fluid, relative to cell surfaces. In mammals, the coordinated beat of motile cilia provides highly specialized functions associated with the movement of luminal contents, as seen with metachronal waves transporting mucus in the respiratory tract. Motile cilia are also present in the male and female reproductive tracts. In the female, wave-like motions of oviductal cilia transport oocytes and embryos toward the uterus. A similar function has been assumed for motile cilia in efferent ductules of the male—i.e., to transport immotile sperm from rete testis into the epididymis. However, we report here that efferent ductal cilia in the male do not display a uniform wave-like beat to transport sperm solely in one direction, but rather exert a centripetal force on luminal fluids through whip-like beating with continual changes in direction, generating turbulence, which maintains immotile spermatozoa in suspension within the lumen. Genetic ablation of two miRNA clusters ( miR-34b/c and -449a/b/c ) led to failure in multiciliogenesis in murine efferent ductules due to dysregulation of numerous genes, and this mouse model allowed us to demonstrate that loss of efferent duct motile cilia causes sperm aggregation and agglutination, luminal obstruction, and sperm granulomas, which, in turn, induce back-pressure atrophy of the testis and ultimately male infertility.

Background Acute myocardial infarction (AMI) is one of the most common global causes of death. Although considerable progress has been made in AMI diagnosis, there remains an urgent need for novel diagnostic biomarkers for its prevention and treatment. Functional exosomal microRNAs (miRNAs) are recognized as potential biomarkers in many diseases. This study's objective was to identify specific plasma exosomal miRNAs with biomarker potential for early AMI detection. Methods Exosomes from the plasma of 26 coronary artery disease (CAD) patients, 55 AMI patients, and 37 healthy controls were isolated and characterized by transmission electron microcopy (TEM), western blotting, and nanoparticle tracking analysis (NTA). The miRNAs were purified from exosomes, and unique molecular identifier (UMI) small RNA sequencing was performed. The random forest (RF) model was trained to predict potential biomarkers. Results NTA demonstrated that nanoparticle concentration did not change after AMI, while nanoparticle size distribution significantly decreased. The CAD and AMI groups' miRNA expression profiles significantly differed from the healthy group's profile. The RF classifier could be used to distinguish the healthy group from the AMI group, but could not be used to distinguish the CAD group from the other groups, which caused a high classification error rate. Eighteen miRNAs were selected as biomarkers based on their RF classifier significance. The diagnostic accuracy of 18 miRNAs was evaluated using AUC values of 0.93, 0.87, and 0.75 to detect healthy controls, AMI, and CAD, respectively. Conclusions Nanoparticle diameter and the 18 miRNAs may serve as simple and accessible fingerprints for early AMI diagnosis.

Single-cell isoform sequencing can reveal transcriptomic dynamics in individual cells invisible to bulk- and single-cell RNA analysis based on short-read sequencing. However, current long-read single-cell sequencing technologies have been limited by low throughput and high error rate. Here we introduce HIT-scISOseq for high-throughput single-cell isoform sequencing. This method was made possible by full-length cDNA capture using biotinylated PCR primers, and by our novel library preparation procedure that combines head-to-tail concatemeric full-length cDNAs into a long SMRTbell insert for high-accuracy PacBio sequencing. HIT-scISOseq yields > 10 million high-accuracy full-length isoforms in a single PacBio Sequel II 8M SMRT Cell, providing > 8 times more data output than the standard single-cell isoform PacBio sequencing protocol. We exemplified HIT-scISOseq by first studying transcriptome profiles of 4,000 normal and 8,000 injured corneal epitheliums from cynomolgus monkeys. We constructed dynamic transcriptome landscapes of known and rare cell types, revealed novel isoforms, and identified injury-related splicing and switching events that are previously not accessible with low throughput isoform sequencing. HIT-scISOseq represents a high-throughput, cost-effective, and technically simple method to accelerate the burgeoning field of long-read single-cell transcriptomics.

Background and Aims 3-methyl histidine (3MH) is a non-proteinogenic amino acid and an index of muscle breakdown. Subclinical protein malnutrition is an independent association with arterial stiffness. The aim of this study was to evaluate the relationship between serum 3MH levels and carotid-femoral pulse wave velocity (cfPWV) values in chronic hemodialysis patients. Method Fasting blood samples and baseline characteristics were obtained from 136 chronic hemodialysis patients. Serum 3MH was performed with high-performance liquid chromatography and mass spectrometry. Aortic arterial stiffness was defined as cfPWV values >10 m/s according to the ESH-ESC 2013 guidelines. Results Among 110 chronic hemodialysis patients, 45 patients (40.9%) were in the aortic arterial stiffness group. When compared to those in the control group, the aortic arterial stiffness group had high prevalence of diabetes mellitus (p < 0.001), hypertension (p = 0.006), older age (p = 0.002), higher systolic blood pressure (p = 0.016), and lower serum 3MH level (p = 0.001). Multivariable logistic regression analysis of the factors significantly associated with aortic arterial stiffness revealed that serum 3MH levels (odds ratio (OR): 0.791, 95% confidence interval (CI): 0.691–0.906, p = 0.001) was the independent predictor of aortic arterial stiffness in chronic hemodialysis patients. Multivariable forward stepwise linear regression analysis also showed that logarithmically transformed 3MH level (log-3MH, β = -0.322, adjusted R2 change = 0.127, p < 0.001) was an independent predictor of cfPWV values in chronic hemodialysis patients. The area under the receiver-operating characteristic (ROC) curve indicates the diagnostic power of 3MH at predicting aortic stiffness of chronic hemodialysis patients was 0.691 (95% CI: 0.595-0.775, p = 0.0002). Conclusion Serum-free 3MH level is negatively associated with aortic arterial stiffness in chronic hemodialysis patients.

Background and Aims Walking speed test is a usefulness tool for cardiovascular risk stratification in older adults. The inhibition of myostatin in adult significantly increases muscle mass and confers benefits upon measures of performance and metabolism. The present study evaluated the relationship between walking speed test and serum myostatin levels in renal transplant recipients. Method Fasting blood samples were collected from 84 renal transplant recipients. Handgrip strength (HGS) was measured using a Jamar Plus Digital Hand Dynamometer for assessment of muscle strength. Gait speed was measured by walking 6 meters at the usual speed. Gait speed < 1 m/s was defined as low gait speed group according to the European Working Group on Sarcopenia in Older People (EWGSOP) criteria. Serum myostatin levels were measured using a commercial enzyme-linked immunosorbent assay. Results Thirty-one renal transplant recipients (36.9%) had low gait speed, and they included a lower percentage of use of mycophenolate mofetil (p = 0.003), higher percentage of use of steroid (p = 0.037), older age (p = 0.0090, higher body weight (p = 0.044), body mass index (p = 0.017), skeletal muscle index (p = 0.027), serum triglyceride (p = 0.029), glucose (p = 0.007), blood urea nitrogen (p = 0.041), cystatin C (p = 0.015), while lower estimated glomerular filtration rate from serum creatinine (eGFRcre, p = 0.047) and estimated glomerular filtration rate from serum cystatin C (eGFRcys, p = 0.006) compared with renal transplant recipients with normal gait speed. After adjusting for cofounders associated with low gait speed in these patients by multivariable logistic regression analysis, serum myostatin levels (Odds ratio (OR): 0.943, 95% confidence interval (CI): 0.898–0.990, p = 0.018), and mycophenolate mofetil used (OR: 0.199, 95% CI: 0.050–0.795, p = 0.022) were independently associated with low gait speed in renal transplant recipients. The area under the receiver-operating characteristic (ROC) curve indicates the diagnostic power of serum myostatin levels at predicting low gait speed of renal transplant recipients was 0.769 (95% CI: 0.664-0.854, p < 0.001). Multivariable forward stepwise linear regression analysis also showed that serum myostatin levels (β = 0.353, adjusted R2 change: 0.245, p = 0.001) was positively associated with gait speed values in renal transplant recipients. Conclusion In this study, serum myostatin levels is found to be positively correlated with gait speed values and is identified as serum low myostatin levels is associated with low gait speed in renal transplant patients.

Background and Aims Resistin named for its ability to resist insulin action has an important link between obesity, insulin resistance and diabetes. Moreover, additional reports suggest that resistin has a pathogenic role in the development and progression of atherosclerosis and coronary artery disease. Chronic kidney disease (CKD) accelerates atherosclerosis via augmentation of inflammation and perturbation of lipid metabolism. Peripheral arterial occlusive disease (PAOD) is associated with an increased risk of death in CKD patients. The aim of this study was to evaluate the relationship between serum resistin level and PAOD by ankle-brachial index (ABI) in CKD patients. Method Fasting blood samples and baseline characteristics were obtained from 240 patients with CKD (stage 3-5). ABI values were measured using the automated oscillometric method (VaSera VS-1000). ABI values that were < 0.9 were included in the low ABI group. Concentrations of human serum resistin were determined using a commercially available enzyme immunoassay kit. Results Among the 240 CKD patients, 30 of them (12.5%) were in the low ABI group. Compared with patients in the control group, the patients in the low ABI group had higher prevalence of diabetes (p = 0.033), hypertension (p = 0.023), current smoking (p = 0.013), older age (p < 0.001), higher systolic blood pressure (p = 0.013), glucose (p = 0.022) level and resistin (p < 0.001) levels. In addition, the multivariable logistic regression analysis showed that serum resistin levels (Odds ratio [OR]: 1.140, 95% confidence interval [CI]: 1.054-1.234, p = 0.001) and age (OR: 1.102, 95% CI: 1.051-1.156, p < 0.001) were the independently associated with PAOD in CKD patients. The area under the receiver-operating characteristic (ROC) curve predicting PAOD by serum resistin level in CKD patients was 0.699 (95% CI: 0.636-0.756, p = 0.0001). Conclusion In this study, serum resistin level was found to be associated with PAOD in non-dialysis CKD stage 3-5 patients.

BACKGROUND: Breast cancer (BC) is one of the most common cancers with high mortality worldwide. In the present study, through bioinformatics analysis, we aimed to identify new biomarkers to predict the survival rate of BC patients. METHODS: Differentially expressed genes (DEGs) between low- and high-tumor mutation burden (TMB) groups were identified by using The Cancer Genome Atlas (TCGA) dataset and integrated analysis. Gene Ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, and the protein-protein interaction (PPI) network, were applied to predict the function of these above DEGs. Then, the Cox proportional hazard model was developed to screen DEGs. Based on the prognostic signature, survival analysis was used on The Cancer Genome Atlas Breast Invasive Carcinoma (TCGA-BRCA) dataset. Finally, the single-sample gene set enrichment (ssGSEA) analysis was employed to estimate immune cells related to this signature. RESULTS: To create a prognostic signature, 6 DEGs were identified. The results revealed that the survival time of patients with high-risk scores based on the expression of the six-gene signature was dramatically shorter than that of patients with low-risk scores in BC. Furthermore, survival analysis and multivariate cox analysis indicated that the six-gene signature was an independent prognostic factor of BC. Then, we built a nomogram that integrated the clinicopathological factors with the six-gene signature to predict the survival probability of BC patients. We eventually predicted the 20 most vital small molecule drugs by CMap, and Nadolol was considered as the most promising small molecule to treat BC. Moreover, ssGSEA analysis showed that the 6 genes were closely associated with immune cells. CONCLUSIONS: We constructed a six-gene signature associated with TMB that can improve the prognosis prediction and could be seen as a biomarker for BC patients.

Coronavirus disease 2019 (COVID-19) is a global pandemic and has been widely reported; however, a comprehensive systemic review and meta-analysis has not been conducted. We systematically investigated the clinical characteristics of COVID-19 in mainland China to guide diagnosis and treatment. We searched the PubMed, Embase, Scopus, Web of Science, Cochrane Library, bioRxiv, medRxiv, and SSRN databases for studies related to COVID-19 published or preprinted in English or Chinese from January 1 to March 15, 2020. Clinical studies on COVID-19 performed in mainland China were included. We collected primary outcomes including signs and symptoms, chest CT imaging, laboratory tests, and treatments. Study selection, data extraction, and risk of bias assessment were performed by two independent reviewers. Qualitative and quantitative synthesis was conducted, and random-effects models were applied to pooled estimates. This study is registered with PROSPERO (number CRD42020171606). Of the 3624 records identified, 147 studies (20,662 patients) were analyzed. The mean age of patients with COVID-19 was 49.40 years, 53.45% were male, and 38.52% had at least one comorbidity. Fever and cough were the most common symptoms, followed by fatigue, expectoration, and shortness of breath. Most patients with COVID-19 had abnormal chest CT findings with ground glass opacity (70.70%) or consolidation (29.91%). Laboratory findings shown lymphopenia, increased lactate dehydrogenase, increased infection-related indicators, and fibrinolytic hyperactivity. Antiviral therapy, antibiotic therapy, and corticosteroids were administered to 89.75%, 79.13%, and 35.64% of patients, respectively. Most clinical characteristics of COVID-19 are non-specific. Patients with suspected should be evaluated by virological assays and clinically treated.

Gastric cancer (GC) is one of the main causes of the high death rate in the world. But the molecular mechanisms of GC carcinogenesis remain little known. This study aimed to identify novel promising biomarkers of GC and reveal its potential molecular mechanisms by integrating bioinformatics analysis. We screened the overlapped differentially expressed genes (DEGs) of TCGA and several GEO datasets. Among these DEGs, we used protein-protein interactions network analysis to recognize the hub genes. Moreover, functional enrichment analysis including GO and KEGG pathway analysis and gene set enrichment analysis (GSEA) were performed to study the role of DEGs and potential underlying mechanisms of GC. Based on integrated bioinformatics analysis, SERPINH1, COL1A2, COL8A1, COL4A1, COL5A1, COL12A1, and COL1A1 were screened as candidate diagnostic marker genes. In addition, SERPINH1 was identified as a core gene in the regulation of GC development. Furthermore, we confirmed that SERPINH1 could promote the proliferation, migration, and cell cycle of GC cells. Our present study demonstrated that SERPINH1 was a core therapeutic biomarker in the regulation of candidate genes involved in GC progression.

Heart failure arises from diverse cardiovascular diseases, including hypertension, ischemic disease and atherosclerosis, valvular insufficiency, myocarditis, and contractile protein mutations. MicroRNAs are dysregulated in heart failure, but identification of the specific microRNAs involved remains incomplete. Here, we evaluate miR-25 expression in the peripheral blood from healthy, dilated cardiomyopathy (DCM), remote infarct (OMI), hypertensive heart disease (HHD), and HHD resulting in heart failure (HHDF) using q-PCR. Interestingly, we discovered miR-25 expression in humans is initially decreased at the onset of heart failure but is later increased in end-stage heart failure. We also show that overexpression of miR-25 in normal mice causes cardiomyocyte fibrosis and apoptosis. However, inhibition of miR-25 in normal mice led to activate renin-angiotensin system (RAS) and high blood pressure, mild heart dilation. Notably, the miR-25 cluster knock-out mice was also characterized high blood pressure and no obvious cardiac function alteration. RNA sequencing showed the alteration of miR-25 target genes in angomir-treated mice, including the renin secretion signal related gene. In vitro, cotransfection with the miR-25 antagomir repressed luciferase activity from a reporter construct containing the Pde3a and Cacnalc untranslated region. In summary, miR-25 expression during different stages of heart disease, offers a new perspective for the role of miR-25 function in heart failure.

To protect operators and conform to safety standards for human&ndash, machine interactions, the design of collaborative robot arms often incorporates flexible mechanisms and force sensors to detect and absorb external impact forces. However, this approach increases production costs, making the introduction of such robot arms into low-cost service applications difficult. This study proposes a low-cost, sensorless rigid robot arm design that employs a virtual force sensor and stiffness control to enable the safety collision detection and low-precision force control of robot arms. In this design, when a robot arm is subjected to an external force while in motion, the contact force observer estimates the external torques on each joint according to the motor electric current and calculation errors of the system model, which are then used to estimate the external contact force exerted on the robot arm&rsquo, s end-effector. Additionally, a torque saturation limiter is added to the servo drive for each axis to enable the real-time adjustment of joint torque output according to the estimated external force, regulation of system stiffness, and achievement of impedance control that can be applied in safety measures and force control. The design this study developed is a departure from the conventional multisensor flexible mechanism approach. Moreover, it is a low-cost and sensorless design that relies on model-based control for stiffness regulation, thereby improving the safety and force control in robot arm applications.

The aim of the present study was to research the mechanism of action of microRNA‑144 (miR‑144) in colorectal cancer (CRC) and its role in tumor progression. It was demonstrated that miR‑144 was downregulated and anoctamin 1 (ANO1) expression was upregulated in CRC. The expression of ANO1 was negatively associated with that of miR‑144 in CRC. The present study indicated that upregulated expression of ANO1 was associated with poor differentiation and advanced tumor‑node‑metastasis stage. It was verified that upregulation of ANO1 expression activated the epidermal growth factor receptor/extracellular signal‑regulated kinase signaling pathway. It was also demonstrated that miR‑144 exerts strong tumor‑inhibiting effects by targeting ANO1. Therefore, miR‑144 may have potential as a prognostic marker or therapeutic target for CRC.

Introduction. Osteoporosis (OP) is a common polygenic disorder in the aging population, and several single nucleotide polymorphisms (SNPs) in the alpha-L-iduronidase (IDUA) gene and patched homolog 1 (PTCH1) gene regulate bone metabolism and affect bone mass. The study aimed at investigating the relationships of rs3755955 and rs6831280 in the IDUA gene and rs28377268 in the PTCH1 gene with bone mineral density (BMD), bone turnover markers (BTMs), and fractures in the elderly Chinese subjects with OP. Materials and Methods. A cohort of 328 unrelated senile osteoporosis (SOP) patients with or without osteoporotic fractures was recruited. rs3755955, rs6831280, and rs28377268 polymorphisms were identified using SNaPshot technology. BTM levels were determined by electrochemiluminescence (ECL). Bone mineral densities (BMDs) at the lumbar spine (LS) and proximal femur sites were measured by dual-energy X-ray absorptiometry (DEXA) in all subjects. The Hardy-Weinberg equilibrium (HWE) test was performed. HWE P values and comparisons of genotype frequencies were estimated using the chi-square test. Analysis of covariance (ANCOVA) adjusted for confounding factors was performed to investigate associations of SNPs with BMDs and BTMs in subgroups. Results. The chi-square test indicated that genotype distributions in the control group conformed to HWE (P>0.05). The distributions of allele and genotype frequencies of rs6831280 between fracture and osteoporotic participants were significantly different (P-allele=0.002 and P-genotype=0.012, respectively). Concerning rs6831280, ANCOVA found BMDs at LS 2-4 (L2-4) and total hip (TH) among the study subjects suffering from SOP with GA genotype were lower than in those carrying GG or AA (P-L2-4=0.004 and P-TH=0.027, respectively). Conclusions. IDUA rs6831280 is associated with BMDs at L2-4 and TH in the elderly Chinese population with SOP and may serve as a marker for the genetic susceptibility to osteoporotic fractures.

Background Accumulating evidence has demonstrated that microRNAs (miRNAs) are associated with tumorigenesis. miR-216b can play a vital role in the genesis and development of gastric cancer (GC), and its molecular mechanisms require further elucidation. Methods The biological effects of miR-216b in GC cells were investigated by MTT, transwell assays, and cell cycle. Western blot and luciferase assay were performed to demonstrate the direct binding of miR-216b on PXN 3′UTR. Furthermore, MTT, colony formation assays, transwell assays, and flow cytometry analysis, as well as xenograft mice model, were used to measure the effects of miR-216b-PXN on GC cell proliferation, migration, and invasion indicated by in vitro and in vivo. Results Our results showed that miR-216b acted as a tumor suppressor in GC progression. miR-216b overexpression suppressed GC cell proliferation, migration, and invasion in vitro. Luciferase reporter assays identified paxillin (PXN) as a novel target gene of miR-216b. PXN overexpression could partially rescue miR-216b-induced the inhibitory effects in GC cells. Besides, overexpression of miR-216b contributed to the activation of PI3K/AKT signaling via partly regulating PXN in GC cells. Conclusions The above results showed that miR-216b could offer a novel therapeutic avenue by targeting PXN in GC.

Background Environmental toxicants such as DDT have been shown to induce the epigenetic transgenerational inheritance of disease (e.g., obesity) through the germline. The current study was designed to investigate the DDT-induced concurrent alterations of a number of different epigenetic processes including DNA methylation, non-coding RNA (ncRNA) and histone retention in sperm. Methods Gestating females were exposed transiently to DDT during fetal gonadal development, and then, the directly exposed F1 generation, the directly exposed germline F2 generation and the transgenerational F3 generation sperm were investigated. Results DNA methylation and ncRNA were altered in each generation sperm with the direct exposure F1 and F2 generations being predominantly distinct from the F3 generation epimutations. The piRNA and small tRNA were the most predominant classes of ncRNA altered. A highly conserved set of histone retention sites were found in the control lineage generations which was not significantly altered between generations, but a large number of new histone retention sites were found only in the transgenerational generation DDT lineage sperm. Conclusions Therefore, all three different epigenetic processes were concurrently altered as DDT induced the epigenetic transgenerational inheritance of sperm epimutations. The direct exposure generations sperm epigenetic alterations were distinct from the transgenerational sperm epimutations. The genomic features and gene associations with the epimutations were investigated to help elucidate the integration of these different epigenetic processes. Observations demonstrate all three epigenetic processes are involved in transgenerational inheritance. The different epigenetic processes appear to be integrated in mediating the epigenetic transgenerational inheritance phenomenon. Electronic supplementary material The online version of this article (10.1186/s13072-018-0178-0) contains supplementary material, which is available to authorized users.

This paper aims at developing a highly efficient walking scheme for compass-like biped robots with feet. Towards this goal, a latch attached to the hip is introduced to lock the hip joint when the hip joint retracts to the desired value and spring devices are installed in the ankle joints to reduce the inputs of the ankles. Moreover, the stance foot allowed to rotate about the toe is considered. With these setups, a highly efficient limit-cycle walking gait is obtained by using the method of Poincaré shooting with the objective of minimizing the dimensionless specific mechanical cost of transport. However, this limit-cycle walking gait is unstable. To stabilize it, a new control strategy is proposed based on transverse coordinate linearization on moving Poincaré sections. A series of numerical simulations show that such a robot model under the proposed control law can walk stably and efficiently.

Mammalian sperm are carriers of not only the paternal genome, but also the paternal epigenome in the forms of DNA methylation, retained histones and noncoding RNAs. Although paternal DNA methylation and histone retention sites have been correlated with protein-coding genes that are critical for preimplantation embryonic development, physiological evidence of an essential role of these epigenetic marks in fertilization and early development remains lacking. Two miRNA clusters consisting of five miRNAs (miR-34b/c and miR-449a/b/c) are present in sperm, but absent in oocytes, and miR-34c has been reported to be essential for the first cleavage division in vitro. Here, we show that both miR-34b/c- and miR-449-null male mice displayed normal fertility, and that intracytoplasmic injection of either miR-34b/c- or miR-449-null sperm led to normal fertilization, normal preimplantation development and normal birth rate. However, miR-34b/c and miR-449 double knockout (miR-dKO) males were infertile due to severe spermatogenic disruptions and oligo-astheno-teratozoospermia. Injection of miR-dKO sperm into wild-type oocytes led to a block at the two-pronucleus to zygote transition, whereas normal preimplantation development and healthy pups were obtained through injection of miR-dKO round spermatids. Our data demonstrate that miR-34b/c and miR-449a/b/c are essential for normal spermatogenesis and male fertility, but their presence in sperm is dispensable for fertilization and preimplantation development.

Quantitative analyses of small RNAs at the single-cell level have been challenging because of limited sensitivity and specificity of conventional real-time quantitative PCR methods. A digital quantitative PCR (dqPCR) method for miRNA quantification has been developed, but it requires the use of proprietary stem-loop primers and only applies to miRNA quantification. Here, we report a microfluidics-based dqPCR (mdqPCR) method, which takes advantage of the Fluidigm BioMark HD system for both template partition and the subsequent high-throughput dqPCR. Our mdqPCR method demonstrated excellent sensitivity and reproducibility suitable for quantitative analyses of not only miRNAs but also all other small RNA species at the single-cell level. Using this method, we discovered that each sperm has a unique miRNA profile.

Cyclic nucleotide-gated channels (CNGCs) have been implicated in diverse aspects of plant growth and development, including responses to biotic and abiotic stress, as well as pollen tube growth and fertility. Here, genetic evidence identifies CNGC16 in Arabidopsis (Arabidopsis thaliana) as critical for pollen fertility under conditions of heat stress and drought. Two independent transfer DNA disruptions of cngc16 resulted in a greater than 10-fold stress-dependent reduction in pollen fitness and seed set. This phenotype was fully rescued through pollen expression of a CNGC16 transgene, indicating that cngc16-1 and 16-2 were both loss-of-function null alleles. The most stress-sensitive period for cngc16 pollen was during germination and the initiation of pollen tube tip growth. Pollen viability assays indicate that mutant pollen are also hypersensitive to external calcium chloride, a phenomenon analogous to calcium chloride hypersensitivities observed in other cngc mutants. A heat stress was found to increase concentrations of 3′,5′-cyclic guanyl monophosphate in both pollen and leaves, as detected using an antibody-binding assay. A quantitative PCR analysis indicates that cngc16 mutant pollen have attenuated expression of several heat-stress response genes, including two heat shock transcription factor genes, HsfA2 and HsfB1. Together, these results provide evidence for a heat stress response pathway in pollen that connects a cyclic nucleotide signal, a Ca2+-permeable ion channel, and a signaling network that activates a downstream transcriptional heat shock response.

Nanoscale Ferric Oxide was prepared from natural hematite and characterized. Using it as catalyst, methylene blue-simulated wastewater was treated by photocatalytic degradation with high-voltage mercury lamp and sunlight as excitation light source. Main factors, including the preparation conditions and dosage of ferric oxide, pH value, reaction time and initial concentration of simulated wastewater, and their influence to treatment effect were discussed. Test results showed that at a pulverization time of 1.5h, calcination time of 2h at 500°C, initial methylene blue (MB) concentration of 20mg/L, pH=2 and a ferric oxide dosage of 0.01g/30ml, for both high-voltage mercury lamp and sunlight, MB wastewater was degraded effectively in lab-scale experiment; after 5h’s radiation, MB concentrations were reduced from 20mg/L to 0.51mg/L and 9.18mg/L respectively. With sunlight as the radiation light source, an enlarged experiment was done on a custom-built device, and MB concentration was reduced from 20mg/L to 0.11mg/L, which was significantly better than treatment results from lab-scale experiments and UV radiation. MB photocatatytic degradation reactions at different initial concentrations were in accordance with Lagergren’s pseudo-first-order kinetic equation. Spectral analysis of degradation products showed that MB molecules were degraded to inorganic ions.

A novel 2009 influenza A (H1N1) virus was transmitted from humans to humans worldwide. The live attenuated monovalent A H1N1 vaccine (LAMV) for intranasal administration has shown promising immunogenicity and safety in clinical trials and for human use, but the experimental data based on LAMV is incomplete. In this study, using reverse genetic technology, we produced a cold-adapted (ca), live attenuated BJ/AA ca that contained hemagglutinin (HA) and neuraminidase (NA) genes from a 2009 pandemic A H1N1 isolate, A/Beijing/501/2009 virus (BJ501), and the remaining six internal gene segments from the cold-adapted influenza H2N2 A/Ann Arbor/6/60 virus (AA virus). BJ/AA ca exhibited phenotypes of temperature sensitivity (ts), ca, and attenuation (att). The candidate BJ/AA ca was immunogenic in mice and induced strong mucosal secretory IgA (sIgA) in the respiratory tract. Two dosages of intranasal immunization induced robust HI antibodies and offered efficient protection against challenge by the wild-type (wt) 2009 pandemic A H1N1 (A/Beijing/501/2009 or A/California/07/2009) in mice and ferrets. These results support the evaluation of this vaccine made from a wt strain isolated in China for clinical trials.

The avian H5N1 influenza virus has the potential to cause a new pandemic. The increasing number of recent outbreaks of highly pathogenic avian influenza H5N1 in birds and humans emphasizes the urgent need to develop a potent H5N1 vaccine. Here, we studied the immunogenicity and protective effect of a vaccine prepared from H5N1 inactivated whole virus. This vaccine was intranasally co-administered in mice with phosphate buffered saline, recombinant cholera toxin B subunit (rCTB), cholera toxin (CT), rCTB containing a trace amount of holotoxin (rCTB/CT), polyinosinic:polycytidylic acid double-stranded RNA (polyI:C), or MF59 as an adjuvant. Intranasal administration of H5N1 inactivated whole virus vaccine with rCTB, CT, rCTB/CT, polyI:C, and MF59 elicited an immunological response with both secretory IgA (sIgA) in nasal, lung, and vaginal lavage, and IgG antibody in serum, showing protective immunity against lethal H5N1 infection. Cross-clade protection was also observed in animals immunized with a vaccine derived from Anhui/01/2005(H5N1) with rCTB, CT, rCTB/CT, polyI:C, or MF59 as adjuvants that were subsequently challenged with the A/OT/SZ/097/03 influenza strain.

How oncogenic virus rewires glucose flux to regulate cell motility is not well-understood. Epstein-Barr virus (EBV) infection of preinvasive nasopharyngeal epithelium is believed to be an essential initiation event during nasopharyngeal carcinoma pathogenesis. Here we demonstrate that EBV-encoded LMP1 enhances glycolysis thereby accelerating cell mobility of nasopharyngeal epithelial (NPE) cells. Activation of IGF1/mTORC2/AKT/PDHE1α signaling which mediates Histone 3 acetylation is involved in cell motility in EBV-infected NPE cells. Blocking the glucose uptake by pharmacological inhibitors effectively suppress glycolysis as well as the cell motility, linking the glucose metabolism to cell movement. We further show that mTORC2/AKT mediated phosphorylation of pyruvate decarboxylase kinase 1 (PDHK1) and its substrate, pyruvate decarboxylase E1α (PDHE1α) component plays a significant role in cell motility. PDHE1α knockdown effectively blocks LMP1-induced cell motility while overexpression of its phosphomimetic mutant S293A but not S293D inhibits LMP1-induced cell motility, supporting the involvement of PDHE1α in enhanced cell motility is depended on Ser293 phosphorylation. Furthermore, PDHE1α translocation into nucleus enhances Histone 3 acetylation and its binding to the promoter region of snail to enhance cell motility. Finally, we find that LMP1 increases the mRNA expression and the secretion of the ligand IGF1, which promotes phosphorylation of IGF1R as the key regulator for LMP1-induced mTORC2/AKT activation. In summary, this study provides evidences of involvement of EBV infection in metabolic reprogramming of NPE cells to promote cell motility involving activation of mTORC2/AKT/PDHE1α. Acknowledgment: This project was supported by funding from the Research Grants Council, Hong Kong: General Research Fund (17120814, 17161116, 17111516, 171110315), Area of Excellence (AoE/M-06/08), Theme-Based Research Scheme (T12-401/13-R), Collaborative Research Fund (Project reference: C7027-16G); Health and Medical Research Fund of Hong Kong (12110782, 13120872, 04151726); and CRCG, University of Hong Kong, Hong Kong. Citation Format: Jun Zhang, Weitao Lin, Chi Man Tsang, Teng Fei Liu, Wing Chong Tang, Yim Ling Yip, Wen Deng, Lin Jia, Sai Wah Tsao. Epstein Barr virus-encoded LMP1 reprograms glucose metabolism to enhance cell motility in nasopharyngeal epithelial cell [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3079.

Although it is believed that mammalian sperm carry small noncoding RNAs (sncRNAs) into oocytes during fertilization, it remains unknown whether these sperm-borne sncRNAs truly have any function during fertilization and preimplantation embryonic development. Germline-specific Dicer and Drosha conditional knockout (cKO) mice produce gametes (i.e., sperm and oocytes) partially deficient in miRNAs and/or endo-siRNAs, thus providing a unique opportunity for testing whether normal sperm (paternal) or oocyte (maternal) miRNA and endo-siRNA contents are required for fertilization and preimplantation development. Using the outcome of intracytoplasmic sperm injection (ICSI) as a readout, we found that sperm with altered miRNA and endo-siRNA profiles could fertilize wild-type (WT) eggs, but embryos derived from these partially sncRNA-deficient sperm displayed a significant reduction in developmental potential, which could be rescued by injecting WT sperm-derived total or small RNAs into ICSI embryos. Disrupted maternal transcript turnover and failure in early zygotic gene activation appeared to associate with the aberrant miRNA profiles in Dicer and Drosha cKO spermatozoa. Overall, our data support a critical function of paternal miRNAs and/or endo-siRNAs in the control of the transcriptomic homeostasis in fertilized eggs, zygotes and 2-cell embryos. Given that supplementation of sperm RNAs enhances both the developmental potential of preimplantation embryos and the live birth rate, it may represent a novel means to improve the success rate of assisted reproductive technologies in fertility clinics.

Nonsense-mediated mRNA decay (NMD) represents a highly conserved RNA surveillance mechanism through which mRNA transcripts bearing premature termination codons (PTCs) are selectively degraded to maintain transcriptomic fidelity in the cell. Numerous in vitro studies have demonstrated the importance of the NMD pathway; however, evidence supporting its physiological necessity has only just started to emerge. Here, we report that ablation of Upf2, which encodes a core NMD factor, in murine embryonic Sertoli cells (SCs) leads to severe testicular atrophy and male sterility owing to rapid depletion of both SCs and germ cells during prepubertal testicular development. RNA-Seq and bioinformatic analyses revealed impaired transcriptomic homeostasis in SC-specific Upf2 knockout testes, characterized by an accumulation of PTC-containing transcripts and the transcriptome-wide dysregulation of genes encoding splicing factors and key proteins essential for SC fate control. Our data demonstrate an essential role of UPF2-mediated NMD in prepubertal SC development and male fertility.

As a member of the large Ran-binding protein family, Ran-binding protein 9 (RANBP9) has been suggested to play a critical role in diverse cellular functions in somatic cell lineages in vitro, and this is further supported by the neonatal lethality phenotype in Ranbp9 global knockout mice. However, the exact molecular actions of RANBP9 remain largely unknown. By inactivation of Ranbp9 specifically in testicular somatic and spermatogenic cells, we discovered that Ranbp9 was dispensable for Sertoli cell development and functions, but critical for male germ cell development and male fertility. RIP-Seq and proteomic analyses revealed that RANBP9 was associated with multiple key splicing factors and directly targeted >2,300 mRNAs in spermatocytes and round spermatids. Many of the RANBP9 target and non-target mRNAs either displayed aberrant splicing patterns or were dysregulated in the absence of Ranbp9. Our data uncovered a novel role of Ranbp9 in regulating alternative splicing in spermatogenic cells, which is critical for normal spermatogenesis and male fertility., Author Summary Male fertility depends on successful production of functional sperm. Sperm are produced through spermatogenesis, a process of male germ cell proliferation and differentiation in the testis. Most of the genes involved in spermatogenesis are transcribed and processed into multiple isoforms, which are mainly achieved through alternative splicing. The testis-specific transcriptome, characterized by male germ cell-specific alternative splicing patterns, has been shown to be essential for successful spermatogenesis. However, how these male germ cells-specific alternative splicing events are regulated remains largely unknown. Here, we report that RANBP9 is involved in alternative splicing events that are critical for male germ cell development, and dysfunction of RANBP9 leads to disrupted spermatogenesis and compromised male fertility.

High tibial osteotomy has been generally accepted as a useful treatment for unicompartment osteoarthritis of the knee to improve a patient's quality of life. Few studies have examined the outcome from the patient's perspective. A survival analysis of one to twenty-one years was conducted on 67 knees including analysis of the patients' satisfaction level with this procedure. Cumulative survival probability of 89.5% at 5 years, 74.7% at 10 years and 66.9 % for 15 and 20 years was reported. Ninety-one percent of patients had improvement in pain score and would choose to have this surgery again. Average patient satisfaction level was 75.5%. Forty-eight percent of patients were able to perform at a higher level of activity than before surgery, although none was able to perform at the level prior to the onset of knee pathology. It was concluded that this procedure was able to improve a patient's quality of life and achieved high satisfaction among patients. There is an increasing role of high tibial osteotomy as an adjunct to an autologous chondrocyte implantation procedure.

Significance Most of the single miRNA gene knockouts display no developmental phenotype. Here, we report that simultaneous inactivation of two functionally overlapping miRNAs, miR-34b/c and miR-449, led to a sexually dimorphic partial perinatal lethality, growth retardation and sterility. Multiple underlying developmental defects, including underdevelopment of the basal forebrain structures, a lack of motile cilia in trachea and oviduct, severely disrupted spermatogenesis and oligoasthenoteratozoospermia, result from the dysregulation of ∼240 target genes that are mainly involved in three major cellular functions, including cell fate control, brain development and microtubule dynamics. This study provides physiological evidence demonstrating an essential role of miR-34b/c and miR-449 in normal brain development, motile ciliogenesis and spermatogenesis.

Pachytene piRNAs are MIWI-/MILI-bound small RNAs abundantly expressed in pachytene spermatocytes and round spermatids in adult mouse testes. Miwi knockout (KO) male mice are sterile due to spermiogenic arrest. In Caenorhabditis elegans, sperm-borne piRNAs appear to have an epigenetic role during fertilization and development because progeny of individuals with piRNA-deficient gametes display a progressive loss of fertility after several generations. In mice, it remains unknown whether pachytene piRNA-deficient round spermatids can produce offspring, and whether the progeny of Miwi mutants also exhibits transgenerational, progressive fertility loss. Here, we report that Miwi KO round spermatids could fertilize both wild-type (WT) and Miwi KO oocytes through round spermatid injection, and could produce healthy and fertile offspring despite the global downregulation of both MIWI-/MILI-bound pachytene piRNAs. Progeny of ROSI-derived heterozygotes, both male and female, displayed normal fertility for at least three generations when bred with either WT or Miwi KO females. Our data indicate that aberrant MIWI-/MILI-bound pachytene piRNA profiles in spermatids do not affect fertilization, early embryonic development, or fertility of the offspring, suggesting that pachytene piRNAs might not be required for paternal transgenerational epigenetic inheritance in mice.

The paper aims at solving the walking control problem of a compass-like biped robot with underactuated ankle on the level ground or even uphill environment. The compass-like biped robot is equipped with a constraint mechanism to lock the hip angle when the swing leg retracts to a desired angle. For this system, an angular momentum based control is presented in order to make the biped robot walk on little downhill slope or even uphill. Existence conditions of limit cycle under angular momentum based control are presented. They can be used to determine whether there exists a gait on the slope or not. Furthermore, we apply the method of Poincare return map to analyze the stability property of the gait with angular momentum based control. Finally, an event-based control is adopted to make the walking gait of compass-like biped robot asymptotically stable.

BackgroundThe increase in recent outbreaks and unpredictable changes of highly pathogenic avian influenza (HPAI) H5N1 in birds and humans highlights the urgent need to develop a cross-protective H5N1 vaccine. We here report our development of a multiple-clade H5N1 influenza vaccine tested for immunogenicity and efficacy to confer cross-protection in an animal model.Methodology/principal findingsMice received two doses of influenza split vaccine with oil-in-water emulsion adjuvant SP01 by intranasal administration separated by two weeks. Single vaccines (3 µg HA per dose) included rg-A/Vietnam/1203/2004(Clade 1), rg-A/Indonesia/05/2005(Clade 2.1), and rg-A/Anhui/1/2005(Clade 2.3.4). The trivalent vaccine contained 1 µg HA per dose of each single vaccine. Importantly, complete cross-protection was observed in mice immunized using trivalent vaccine with oil-in-water emulsion adjuvant SP01 that was subsequently challenged with the lethal A/OT/SZ/097/03 influenza strain (Clade 0), whereas only the survival rate was up to 60% in single A/Anhui/1/2005 vaccine group.Conclusion/significanceOur findings demonstrated that the multiple-clade H5N1 influenza vaccine was able to elicit a cross-protective immune response to heterologous HPAI H5N1 virus, thus giving rise to a broadly cross-reactive vaccine to potential prevention use ahead of the strain-specific pandemic influenza vaccine in the event of an HPAI H5N1 influenza outbreak. Also, the multiple-clade adjuvanted vaccine could be useful in allowing timely initiation of vaccination against unknown pandemic virus.

BACKGROUND: The 2009 influenza pandemic affected people in almost all countries in the world, especially in younger age groups. During this time, the debate over whether to use corticosteroid treatment in severe influenza H1N1 infections patients resurfaced and was disputed by clinicians. There is an urgent need for a susceptible animal model of 2009 H1N1 infection that can be used to evaluate the pathogenesis and the therapeutic effect of corticosteroid treatment during infection. METHODOLOGY/PRINCIPAL FINDINGS: We intranasally inoculated two groups of C57BL/6 and BALB/c mice (using 4- or 6-to 8-week-old mice) to compare the pathogenesis of several different H1N1 strains in mice of different ages. Based on the results, a very susceptible 4-week-old C57BL/6 mouse model of Beijing 501 strain of 2009 H1N1 virus infection was established, showing significantly elevated lung edema and cytokine levels compared to controls. Using our established animal model, the cytokine production profile and lung histology were assessed at different times post-infection, revealing increased lung lesions in a time-dependent manner. In additional,the mice were also treated with dexamethasone, which significantly improved survival rate and lung lesions in infected mice compared to those in control mice. Our data showed that corticosteroid treatment ameliorated acute lung injury induced by the 2009 A/H1N1 virus in mice and suggested that corticosteroids are valid drugs for treating 2009 A/H1N1 infection. CONCLUSIONS/SIGNIFICANCE: Using the established, very susceptible 2009 Pandemic Influenza A (H1N1) mouse model, our studies indicate that corticosteroids are a potential therapeutic remedy that may address the increasing concerns over future 2009 A/H1N1 pandemics.

Background Conventional transabdominal ultrasound usually fails to visualize parts of the ureter or extrahepatic bile duct covered by bowel gas. In this study, we propose a new method for gaining acoustic access to the ureters and extrahepatic bile duct to help determine the nature of obstruction to these structures when conventional transabdominal ultrasound fails. Methods The normal saline retention enema method, that is, using normal saline-filled colons to gain acoustic access to the bilateral ureters and extrahepatic bile duct and detecting the lesions with transabdominal ultrasonic diagnostic apparatus, was applied to 777 patients with obstructive lesions, including 603 with hydroureter and 174 with dilated common bile duct, which were not visualized by conventional ultrasonography. The follow-up data of all the patients were collected to verify the results obtained by this method. Results Of the 755 patients who successfully finished the examination after normal saline retention enema (the success rate of the enema is about 98%), the nature of obstruction in 718 patients was determined (the visualizing rate is approximately 95%), including 533 with ureteral calculus, 23 with ureteral stricture, 129 with extrahepatic bile duct calculus, and 33 with common bile duct tumor. Conclusions Colons filled fully with normal saline can surely give acoustic access to the bilateral ureters and extrahepatic bile duct so as to determine the nature of obstruction of these structures when conventional transabdominal ultrasound fails.

Oil-in-water emulsions are potent human adjuvants commonly used in effective pandemic influenza vaccines; however, such emulsions that can induce both Th1-biased systemic immune responses and strong mucosal immune responses via an easy method of administration are lacking. To address this need for new adjuvants, we developed a novel oil/water emulsion, SPO1, which allows convenient mucosal immunization via an intranasal spray as well as by parenteral routes. Our report shows that SPO1 was able to boost up immunological resistance by inducing effective mucosal and serum antibodies, and the immune response was polarized to a Th1 pattern, as demonstrated by high IgG2α antibody levels and interferon-gamma production by splenocytes from intranasally (i.n.) immunized mice. Up-regulation of co-stimulatory and antigen-presenting molecules on dendritic cells was also observed in vivo after i.n. immunization, suggesting a possible mechanism for the adjuvant effects of SPO1. Another explanation may simply be a depot of antigen at the immunization site, as evidenced by in vivo imaging of i.n. immunized mice. In conclusion, our results demonstrate that a novel oil/water emulsion, SPO1, is a potent Th1 adjuvant for use in influenza and other vaccines, as it induces strong mucosal and systemic immune responses.

The 2009 flu pandemic involved the emergence of a new strain of a swine-origin H1N1 influenza virus (S-OIV H1N1) that infected almost every country in the world. Most infections resulted in respiratory illness and some severe cases resulted in acute lung injury. In this report, we are the first to describe a mouse model of S-OIV virus infection with acute lung injury and immune responses that reflect human clinical disease. The clinical efficacy of the antiviral oseltamivir (Tamiflu) administered in the early stages of S-OIV H1N1 infection was confirmed in the mouse model. Moreover, elevated levels of IL-17, Th-17 mediators and IL-17-responsive cytokines were found in serum samples of S-OIV-infected patients in Beijing. IL-17 deficiency or treatment with monoclonal antibodies against IL-17-ameliorated acute lung injury induced by the S-OIV H1N1 virus in mice. These results suggest that IL-17 plays an important role in S-OIV-induced acute lung injury and that monoclonal antibodies against IL-17 could be useful as a potential therapeutic remedy for future S-OIV H1N1 pandemics.

The continued spread of a highly pathogenic avian influenza (HPAI) H5N1 virus among poultry and wild birds has posed a potential threat to human public health. An influenza pandemic happens, when a new subtype that has not previously circulated in humans emerges. Almost all of the influenza pandemics in history have originated from avian influenza viruses (AIV). Birds are significant reservoirs of influenza viruses. In the present study, we performed a survey of avian influenza virus in ostriches and H5N1 virus (A/Ostrich/SuZhou/097/03, China097) was isolated. This H5N1 virus is highly pathogenic to both chickens and mice. It is also able to replicate in the lungs of, and to cause death in, BALB/c mice following intranasal administration. It forms plaques in chicken embryo fibroblast (CEF) cells in the absence of trypsin. The hemagglutinin (HA) gene of the virus is genetically similar to A/Goose/Guangdong/1/96(H5N1) and belongs to clade 0. The HA sequence contains multiple basic amino acids adjacent to the cleavage site, a motif associated with HPAI viruses. More importantly, the existence of H5N1 isolates in ostriches highlights the potential threat of wild bird infections to veterinary and public health.

The novel influenza A (H1N1) 2009 virus has emerged to cause the first pandemic of the twenty-first century. Disease outbreaks caused by the influenza A (H1N1) virus have prompted concerns about the potential for a pandemic and have driven the development of vaccines against this subtype of influenza A. In this study, we developed a monovalent influenza A (H1N1) split vaccine and evaluated its effects in BALB/c mice. Mice were immunized subcutaneously with 2 doses of the vaccine containing hemagglutinin (HA) alone or HA plus an aluminum hydroxide (Al(OH)(3)) adjuvant. Immunization with varying doses of HA (3.75, 7.5, 15, 30, 45 or 60 microg) was performed to induce the production of neutralizing antibodies. The vaccine elicited strong hemagglutination inhibition (HI) and microneutralization, and addition of the adjuvant augmented the antibody response. A preliminary safety evaluation showed that the vaccine was not toxic at large doses (0.5 ml containing 60 microg HA+600 microg Al(OH)(3) or 60 microg HA). Moreover, the vaccine was found to be safe at a dose of 120 microg HA+1200 microg Al(OH)(3) or 120 microg HA in 1.0 ml in rats. In conclusion, the present study provides support for the clinical evaluation of influenza A (H1N1) vaccination as a public health intervention to mitigate a possible pandemic. Additionally, our findings support the further evaluation of the vaccine used in this study in primates or humans.