Separation of spring viraemia of carp virus from large-volume samples using immunomagnetic beads.

A method for separation of spring viraemia of carp virus (SVCV) from large-volume samples using immunomagnetic beads (IMBs) coated with a polyclonal antibody against SVCV was developed. The optimum amount of IMBs was 2 mg in 100 mL. After IMB treatment, the detection limit of SVCV in reverse transcription quantitative PCR (RT-qPCR) was 103 times the 50% tissue culture infectious dose per mL in 100-mL samples. The concentration of viral RNA extracted from SVCV that had been separated using IMBs was 5.18 × 103-fold higher than that of the unseparated SVCV. When fish samples were tested, the concordance rates of the IMBs/RT-qPCR and RT-qPCR were 100% and 67.5%, respectively.

Establishment of a fluorescent PCR melting curve method for detecting asthma susceptibility using gene SNP typing.

Objective: To develop a detection method for single nucleotide polymorphisms (SNPs) of bronchial asthma (BA) susceptibility genes (IL-13, IL-33, and GSDMA) based on fluorescence PCR melting curves.Methods: Peripheral blood samples from 33 patients with BA were collected. DNA was extracted, and positive plasmids were constructed. Probes and primers for fluorescence polymerase chain reaction (PCR) were designed according to IL-13, IL-33, and GSDMA sequences, and the SNPs were separately detected by gene sequencing and fluorescence PCR melting curve.Results: The system was successfully divided into 3 SNPs, including IL-13, IL-33, and GSDMA, and a comparison of sequencing methods showed that the results were completely consistent. The lowest detection limit was 1 ng/reaction, the sensitivity and specificity were 100%, and this method had high repeatability (CV = 2.8%).Conclusion: The fluorescence PCR melting curve method is suitable for the rapid and accurate classification of SNPs. The method is economical, simple, and efficient, and is suitable for the screening of the susceptible gene SNPs in a large-scale population of patients with BA.

The Associations of Genetic Variants in E-cadherin Gene With Clinical Outcome of Epithelial Ovarian Cancer.

OBJECTIVE: The E-cadherin protein plays major roles in tumor progression, invasion, and metastasis. Polymorphisms located in the E-cadherin gene (CDH1) may contribute to increased risks of specific cancers. In this study, we evaluated the associations between genetic variants in CDH1 and the clinical outcomes of patients with epithelial ovarian cancer (EOC). MATERIALS AND METHODS: We assessed the -160C/A and -347G/GA polymorphisms in the promoter region, as well as the 3'-UTR +54C/T polymorphism of E-cadherin, in 257 patients with EOC by ligase detection reaction-polymerase chain reaction. RESULTS: Multivariate analysis showed that patients with EOC with the CDH1 -347GA/GA genotype had shorter progression-free survival and overall survival (hazard ratio [HR], 2.16; 95% confidence interval [CI], 1.06-4.40 and HR, 2.06; 95% CI, 1.01-4.19, respectively) compared to those carrying the G/G genotype. Likewise, the patients with the CDH1 -160A/A genotype had a shorter progression-free survival than those with the C/C genotype (HR, 4.12; 95% CI, 1.43-111.88). No significant association was detected between the CDH1 3'-UTR +54C/T polymorphism and survival of the patients with EOC. CONCLUSIONS: The CDH1 -347GA/GA and -160A/A genotypes may be prognostic markers that can help to identify patients at increased risk of invasive/metastatic cancer in northern China.

The clinical characteristics of alcohol-related ocular rupture.

PURPOSE: To evaluate the characteristics and outcomes of drunken patients treated for ocular rupture, and to compare these results to patients injured without alcohol consumption. DESIGN AND METHODS: The medical records of 182 patients with or without alcohol consumption before injury who were treated and followed up because of ocular rupture at the Affiliated Hospital of Weifang Medical University from October 2007 to October 2011 were evaluated retrospectively. The characteristics and outcomes of 45 alcohol-related injury patients were compared with the rest in the cohort. The clinical data included in this study were: anatomic sites and length of the wound, involvement of ocular adnexa injuries, evisceration rate, and final mean visual acuity. RESULTS: Wound locations were significantly different between the alcohol-related group and the non-alcohol-related one. Compared with the non-alcohol-related ocular rupture population, the anatomic sites of the drunken patients were more likely to be located at zone I and zone II (60.0 vs 40.1 %; χ2 = 5.39,P < 0.05). The difference of wound length between the alcohol-related group and the non-alcohol-related one was significant. The alcohol-related patients had a longer wound length (Z = -8.590,P < 0.05). Compared with the non-alcohol population, the alcohol-consuming patients were more likely to suffer adnexa injuries (84.4 vs 59.8 %; χ2 = 5.86,P < 0.05), and had worse final visual acuities (Z = -7.195,P < 0.05). The evisceration rate of the alcohol-related patients was significantly higher than the non-alcohol patients (24.4 vs 9.4 %; χ2 = 6.62,P < 0.05). CONCLUSIONS: Drinking more easily leads to injury of the front part of eyes. Moreover, the drunken patients had a worse visual acuity outcome, longer wound length, higher evisceration rate, and were more prone to endure adnexa injuries. The importance of prevention and education to recognize the hazards of drinking cannot be overemphasized.

Detection of grass carp reovirus (GCRV) with monoclonal antibodies.

Grass carp reovirus (GCRV) is a pathogen that causes hemorrhagic disease of grass carp. It is the most serious infectious disease of carp and causes serious losses of fingerlings of grass carp and black carp. In this study, a recombinant VP4, one of the viral core proteins, was constructed with a histidine tag and expressed at a high level in E. coli, and the expressed protein was mainly found in the form of inclusion bodies. The expressed VP4 protein was recognized by an anti-His-tag monoclonal antibody and goat anti-GCRV serum. Four monoclonal antibodies (16B7, 39E12, 13C3 and 14D1) against the recombinant VP4 protein were produced. These MAbs did not react with any of the tested viruses or fish cells lines in the ELISA tests except GCRV. In western blotting analysis, a protein band was observed when the recombinant VP4 protein of GCRV was used as an antigen, but a 68-kDa band was observed when natural capsid proteins of GCRV were used as antigens. Furthermore, a sandwich ELISA was developed for detection of GCRV. The detection limit of the test was 105 TCID50 of GCRV per mL.

A Large Asymptomatic Polyp Originating From the Nasal Septum: A Case Report.

A neoplasm was found in the left nasal cavity of a 45-year-old woman during electronic laryngoscopy for reflux pharyngitis. She reported experiencing an occasional slight headache in the left parietal region for 1 to 2 years, which she considered a migraine. Electronic laryngoscopy showed a gray, soft, smooth neoplasm in the left nasal meatus, located near the olfactory region blocking the olfactory clef and compressing the left middle turbinate. The neoplasm was resected at endonasal endoscopic surgery. Histological assessments indicated chronic mucus inflammation and cyst formation. This is a rare case because the polyp was large but asymptomatic and originated from nasal septum.

In-situ sequential crystallization of fenofibrate and tristearin - Understanding the distribution of API in particles and stability of solid lipid microparticles from the perspective of crystallization.

In-situ API crystallization in carrier matrices has attracted extensive attention in recent years for its advantages over traditional preparation processes. However, due to the lack of systemic research on molecular self-assembly behaviors, the products obtained by in-situ crystallization suffer from the problems of polymorphic transformation and drug expulsion during storage, limiting its industrial application. This paper investigates the in-situ sequential crystallization behavior of tristearin (SSS) and fenofibrate (FEN), utilizing SSS as the carrier and FEN as the API. It was found that the behavior of mixed crystallization significantly differs from single-component crystallization, including direct formation of stable form of SSS and the rapid crystallization of FEN. During the crystallization process, the melting FEN promotes the movement of SSS molecules, while the sliding of SSS lamellae, in turn, provides a mechanical stimulus to enhance the nucleation of FEN. Based on the observed synergistic crystallization behavior, the distribution and stability of the API within FEN solid lipid microparticles (SLMs) during storage were evaluated, while also examining the stability variations in SLMs formulated at different cooling rates and drug loading concentrations. The findings indicate that the initial nucleated FEN results in a decrease in the surrounding molten FEN and the irregularity of the SSS lamellas, thereby preventing the remaining molten FEN from achieving complete crystallization within a brief period. Due to the compatibility between FEN and SSS, some SSS may blend with the molten FEN, potentially resulting in further crystallization during storage and consequently increasing the risk of drug expulsion.

The promoter of miR-663 is hypermethylated in Chinese pediatric acute myeloid leukemia (AML).

BACKGROUND: There is growing evidence supporting a role for microRNAs (miRNA) as targets in aberrant mechanisms of DNA hypermethylation. Epigenetic silencing of tumor suppressor miRNAs, including miR-663, which has recently been reported to be inactivated by hypermethylation in several cancers, may play important roles in pediatric acute myeloid leukemia (AML). However, expression of miR-663 and its promoter methylation remain status unclear in childhood leukemia. METHODS: Promoter methylation status of miR-663 was investigated by methylation specific PCR (MSP) and bisulfate genomic sequencing (BGS). Transcriptional expression of miR-663 was evaluated by semi-quantitative and real-time PCR, and the relationship between expression of miR-663 and promoter methylation was confirmed using 5-aza-2'-deoxycytidine (5-Aza) demethylation reagent. RESULTS: MiR-663 was aberrantly methylated in 45.5% (5/11) leukemia cell lines; BGS showed that the promoter was significantly methylated in three AML cell lines; methylation of miR-663 was significantly higher in Chinese pediatric AML patients [41.4% (29/70)] compared to normal bone marrow (NBM) control samples [10.0% (3/30)]. These results were confirmed by both BGS and 5-Aza demethylation analysis. In addition, miR-663 transcript expression was significantly lower in AML patients, both with and without miR-663 methylation, compared to controls; however, there were no significant differences in clinical features or French-American-British (FAB) classification between patients with and without miR-663 methylation. CONCLUSIONS: Expression of miR-663 was significantly lower in pediatric AML cells compared to NBM controls; furthermore, a high frequency of miR-663 promoter hypermethylation was observed in both AML cell lines and pediatric AML samples. Inactivation of miR-663 by promoter hypermethylation could be affected by 5-Aza demethylation. These findings suggest that hypermethylation of the miR-663 promoter may be an early event in the development of pediatric AML.

A loop-mediated isothermal amplification method for the detection of members of the genus Ranavirus.

A loop-mediated isothermal amplification (LAMP) method was developed for detection of members of the genus Ranavirus. The optimum reaction mixture contained 2.5 µL of each inner primer, RV-FIP (20 pmol/µL) and RV-BIP (20 pmol/µL), 0.5 µL of each outer primer, RV-F3 (10 pmol/µL) and RV-B3 (10 pmol/µL), 1.25 µL of each loop primer, RV-LF (20 pmol/µL) and RV-LB (20 pmol/µL), 3.5 µL dNTP mix (10 mM each), 8 µL MgSO4 (25 mM), 1 µL of Bst DNA polymerase (8 U/mL, large fragment; New England Biolabs Inc., Beverly, MA, USA), 2.5 µL 10 × supplied buffer, and 1 µL of template DNA in a final volume of 25 µL. The optimum reaction conditions were 63 °C for 60 min. This LAMP method could detect Andrias davidianus iridovirus (ADIV), soft-shelled turtle iridovirus (STIV), and epizootic hematopoietic necrosis virus (EHNV), all of which belong to the genus Ranavirus, but it could not detect other viruses such as koi herpes virus (KHV), channel catfish virus (CCV), infectious spleen and kidney necrosis virus (ISKNV) and white spot syndrome virus (WSSV). The detection limit of the LAMP method was 100 copies of STIV DNA segment, and the sensitivity was 10 times higher than that of the polymerase chain reaction (PCR) assay. The results could be estimated visually by eye when calcein was added.

Analyzing the gene expression profile of pediatric acute myeloid leukemia with real-time PCR arrays.

BACKGROUND: The Real-time PCR Array System is the ideal tool for analyzing the expression of a focused panel of genes. In this study, we will analyze the gene expression profile of pediatric acute myeloid leukemia with real-time PCR arrays. METHODS: Real-time PCR array was designed and tested firstly. Then gene expression profile of 11 pediatric AML and 10 normal controls was analyzed with real-time PCR arrays. We analyzed the expression data with MEV (Multi Experiment View) cluster software. Datasets representing genes with altered expression profile derived from cluster analyses were imported into the Ingenuity Pathway Analysis Tool. RESULTS: We designed and tested 88 real-time PCR primer pairs for a quantitative gene expression analysis of key genes involved in pediatric AML. The gene expression profile of pediatric AML is significantly different from normal control; there are 19 genes up-regulated and 25 genes down-regulated in pediatric AML. To investigate possible biological interactions of differently regulated genes, datasets representing genes with altered expression profile were imported into the Ingenuity Pathway Analysis Tool. The results revealed 12 significant networks. Of these networks, Cellular Development, Cellular Growth and Proliferation, Tumor Morphology was the highest rated network with 36 focus molecules and the significance score of 41. The IPA analysis also groups the differentially expressed genes into biological mechanisms that are related to hematological disease, cell death, cell growth and hematological system development. In the top canonical pathways, p53 and Huntington's disease signaling came out to be the top two most significant pathways with a p value of 1.5E-8 and2.95E-7, respectively. CONCLUSIONS: The present study demonstrates the gene expression profile of pediatric AML is significantly different from normal control; there are 19 genes up-regulated and 25 genes down-regulated in pediatric AML. We found some genes dyes-regulated in pediatric AML for the first time as FASLG, HDAC4, HDAC7 and some HOX family genes. IPA analysis showed the top important pathways for pediatric AML are p53 and Huntington's disease signaling. This work may provide new clues of molecular mechanism in pediatric AML.

RhoA/ROCK may involve in cardiac hypertrophy induced by experimental hyperthyroidism.

In this study, the role of the RhoA/Rho-kinase (RhoA/ROCK)-signaling pathway in cardiovascular dysfunction associated with hyperthyroidism was examined with the use of fasudil, a Rho-kinase inhibitor. Male Spraque-Dawley rats were treated with l-thyroxine (T(4)) alone, T(4) + low-dose fasudil (2 mg/kg/day) or T(4) + high-dose fasudil (10 mg/kg/day) and compared with control animals. Rats in the T(4) group showed an increase in the ratio of heart weight to body weight, which was ameliorated by fasudil at both low and high doses. Morphometric and hemodynamic parameters were also evaluated and confirmed that fasudil attenuated the cardiac hypertrophy induced by T(4). The extent of phosphorylation of the myosin phosphatase targeting subunit was quantified by Western blotting to evaluate the activity of Rho-kinase in the heart tissue. Both Western blotting and reverse transcriptase-polymerase chain reaction analyses revealed enhancement of Rho-kinase and activator protein 1 activity and reduction of c-FLIP(L) expression in the T(4) group, and this response was inhibited by fasudil in a dose-dependent manner. Furthermore, fasudil inhibited apoptosis induced by T(4) as evidenced by the detection of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells and the expressions of bax and bcl-2. These results suggested that the RhoA/ROCK pathway is involved in the cardiac hypertrophy induced by experimental hyperthyroidism. The antagonism of this pathway may thus be useful as an alternative target in the treatment of hyperthyroid heart disease.

Inter-annual variation patterns in the carbon footprint of farmland ecosystems in Guangdong Province, China.

Carbon sequestration in farmland ecosystems is an important link in the world carbon cycle and plays an important role in regional carbon reduction. Guangdong, a major industrial and economic province in China, was used as the study area, and the period 2001-2020 was taken as the study period. The carbon emissions, sequestration, and footprint of farmland ecosystems in Guangdong were estimated using carbon emission factors for agricultural inputs that are closer to the actual situation in China. The study showed that: (1) Carbon average emissions from farmland in Guangdong during the study period was 3.7624 million t a-1, with a balanced overall trend of change, and that nitrogen fertilize applications was the main factor contributing to carbon emissions. (2) The carbon sink capacity of Guangdong farmland ecosystems showed an overall decreasing trend of 10.32%, with an average annual carbon sink of 19.0363 million t a-1. Paddy and sugar cane cultivations were the main factor of carbon sink in farmland. (3) The average annual carbon footprint of Guangdong's farmland ecosystems was 531,100 ha a-1, which was in a carbon surplus. Carbon surplus and footprint showed a decreasing trend year by year. The paper results provide a theoretical basis for the formulation of carbon emission reduction policies and industrial restructuring in Guangdong and provinces with the same industrial structure.

3D printing calcium alginate adsorbents for highly efficient recovery of U(VI) in acidic conditions.

In this study, a three-dimensional (3D) porous calcium alginate (3D CA) scaffold was successfully constructed using a direct-ink-writing-based 3D printing method combined with an in-situ calcium ion cross-linking procedure. The 3D CA contained orderly aligned microstructures with excellent structural robustness and an abundant number of active binding sites. The adsorption experiments verified that 3D CA had a considerably wide pH value (3-10) serving range, but also delivered a significantly higher adsorption capacity for U(VI) (117.3 mg/g at pH = 2.5) under acidic conditions, compared to other previously reported alginate-based porous adsorbents. The adsorption mechanisms originated from the synergistic effect of electrostatic interactions and ion exchange. The 3D CA eluted the adsorbed U(VI) in a strong acid solution through protonation mechanism, facilitating the continued enrichment and recycling of U(VI). In addition, the 3D CA demonstrated good microstructure stability and absorption capacity stability when it was immersed in hydrochloric acid solutions at different concentrations (3.6 × 10-3 to 2 mol/L) for 24 h. Therefore, the 3D CA could be used for the removal and recycling of U(VI) from acidic solutions beyond its wide pH working range, due to its stronger acid stability and higher U(VI) adsorption capacity.

Down-regulation of Toll-like receptor 4 gene expression by short interfering RNA attenuates bone cancer pain in a rat model.

BACKGROUND: This study demonstrates a critical role in CNS innate immunity of the microglial Toll-like receptor 4 (TLR4) in the induction and maintenance of behavioral hypersensitivity in a rat model of bone cancer pain with the technique of RNA interference (RNAi). We hypothesized that after intramedullary injection of Walker 256 cells (a breast cancer cell line) into the tibia, CNS neuroimmune activation and subsequent cytokine expression are triggered by the stimulation of microglial membrane-bound TLR4. RESULTS: We assessed tactile allodynia and spontaneous pain in female Sprague-Dawley (SD) rats after intramedullary injection of Walker 256 cells into the tibia. In a complementary study, TLR4 small interfering RNA(siRNA) was administered intrathecally to bone cancer pain rats to reduce the expression of spinal TLR4. The bone cancer pain rats treated with TLR4 siRNA displayed significantly attenuated behavioral hypersensitivity and decreased expression of spinal microglial markers and proinflammatory cytokines compared with controls. Only intrathecal injection of TRL4 siRNA at post-inoculation day 4 could prevent initial development of bone cancer pain; intrathecal injection of TRL4 siRNA at post-inoculation day 9 could attenuate, but not completely block, well-established bone cancer pain. CONCLUSIONS: TLR4 might be the main mediator in the induction of bone cancer pain. Further study of this early, specific, and innate CNS/microglial response, and how it leads to sustained glial/neuronal hypersensitivity, might lead to new therapies for the prevention and treatment of bone cancer pain syndromes.

Influences of galactose ligand on the uptake of TADF liposomes by HepG2 cells.

Glucose is the main energy substance to drive the physiological events of the cell.. Malignant cells exhibit a much higher rate of glycolysis than healthy cells to relieve the increased needs of energy. The higher metabolic rate induces the over-expression of the Glucose Transporter (GLUT) to transport more glucose into malignant cells. Our research regarded overexpressive GLUT as a target of nanoparticles. Substrate of GLUT galactose conjugated Polyethylene glycol-Distearyl phosphatidyl ethanolamine (PEG-DSPE) as a kind of ligand was selected to modified liposome. Thermally activated delayed fluorescence (TADF) was encapsulated as fluorescent probe to evaluate its abilities of targeting malignant cells, and the results of confocal microscopy and flow cytometry demonstrated that Galactose-PEG-DSPE modified liposome had the stronger efficiency of cellular uptake by HepG2 cells compared with Blank-PEG-DSPE modified liposome. The effect of GLUT1 inhibitor on cellular uptake of Galactose-PEG-DSPE modified liposomes showed that the mechanism might be relative to Warburg effect causing GLUT overexpression.

Antibiotic resistance ofKlebsiella pneumoniae through ß-arrestin recruitment-induced ß-lactamase signaling pathway.

Overuse and misuse of antibiotics leads to rapid evolution of antibiotic-resistant bacteria and antibiotic resistance genes. Klebsiella pneumoniae has become the most common pathogenic bacterium accountable for nosocomial infections due to its high virulence factor and general occurrence of resistance to most antibiotics. The ß-lactamase signaling pathway has been suggested to be involved in antibiotic resistance against ß-lactams in Klebsiella pneumoniae. In the present study, the molecular mechanism of the antibiotic resistance of Klebsiella pneumoniae was investigated and the results indicated involvement of the ß-arrestin recruitment-induced ß-lactamase signaling pathway. Antimicrobial susceptibility of Klebsiella pneumoniae was assessed using automated systems and extended-spectrum ß-lactamase (ESBL) and ß-arrestin expression levels in Klebsiella pneumoniae were analyzed by reverse-transcription quantitative PCR. ß-lactam resistance in Klebsiella pneumoniae was determined using ß-lactam agar screening plates. The results demonstrated that ß-arrestin recruitment was increased in Klebsiella pneumoniae with antibiotic resistance (AR-K.P.) compared with that in the native Klebsiella pneumoniae strain (NB-K.P.). Increased production of ESBL was observed in AR-K.P. after treatment with the ß-lactam penicillin. Of note, inhibition of ß-arrestin recruitment significantly suppressed ESBL expression in AR-K.P. and in addition, genes encoding ß-arrestin and ESBL were upregulated in Klebsiella pneumoniae. Restoration of endogenous ß-arrestin markedly increased antibiotic resistance of Klebsiella pneumoniae to ß-lactam. Knockdown of endogenous ß-arrestin downregulated antibiotic resistance genes and promoted the inhibitory effects of ß-lactam antibiotic treatment on Klebsiella pneumoniae growth. In conclusion, the present study identified that ß-arrestin recruitment was associated with growth and resistance to ß-lactams, which suggested that ß-arrestin regulating ESBL expression may be a potential target for addressing antibiotic resistance to ß-lactams in Klebsiella pneumoniae.

[Clinical application of cone beam computed tomography combined with micro-ultrasound technique in treating three mesial canals in mandibular first molars].

Objective The aim of this study is to investigate the influence of cone beam computed tomography (CBCT) and micro-ultrasound technique for the treatment of three mesial canals in mandibular first molars. The three mesial canals according to Pomeranz's classification were characterized. Methods A total of 75 permanent mandibular first molars for root canal treatment were randomly selected from patients belonging to the age group of 14-60 years. After preparing the access cavity and locating the main canals, the middle mesial canal orifices in all teeth were determined with an endodontic explorer under direct vision (StageⅠ), under magnification with the aid of micro-ultrasound (Stage Ⅱ), and with the combined use of CBCT and micro-ultrasound to remove the dentin wall and calcifications (Stage Ⅲ). Results Middle mesial canals were detected in 4.0%, 18.7%, and 22.7% of the teeth in StagesⅠ-Ⅲ, respectively. Statistical analysis showed significant differences (P<0.05) between StagesⅠand Ⅱ with regard to middle mesial canal detection. The number of Stage Ⅲ was more than that of Stage Ⅱ. The difference between the two stages was no significant. Among the 17 middle mesial canals, "confluent", "fin" and "independent" anatomies were 52.9%, 35.3%, and 11.8%. Conclusion When used with adjunctive aids, including CBCT, micro-ultrasound facilitates dental clinicians in the location and treatment of middle mesial canals.

Classification via Sparse Representation of Steerable Wavelet Frames on Grassmann Manifold: Application to Target Recognition in SAR Image.

Automatic target recognition has been widely studied over the years, yet it is still an open problem. The main obstacle consists in extended operating conditions, e.g.., depression angle change, configuration variation, articulation, and occlusion. To deal with them, this paper proposes a new classification strategy. We develop a new representation model via the steerable wavelet frames. The proposed representation model is entirely viewed as an element on Grassmann manifolds. To achieve target classification, we embed Grassmann manifolds into an implicit reproducing Kernel Hilbert space (RKHS), where the kernel sparse learning can be applied. Specifically, the mappings of training sample in RKHS are concatenated to form an overcomplete dictionary. It is then used to encode the counterpart of query as a linear combination of its atoms. By designed Grassmann kernel function, it is capable to obtain the sparse representation, from which the inference can be reached. The novelty of this paper comes from: 1) the development of representation model by the set of directional components of Riesz transform; 2) the quantitative measure of similarity for proposed representation model by Grassmann metric; and 3) the generation of global kernel function by Grassmann kernel. Extensive comparative studies are performed to demonstrate the advantage of proposed strategy.