B. Reduced protein ABT-888 mw expression of LATS1 in glioma. 1: Strong expression of LATS1 in normal brain; 2: Strong expression of LATS1 in glioma WHO grade-1; 3: Strong expression of LATS1 in glioma WHO grade-2; 4: Weak expression of LATS1 in glioma WHO grade-3. 5. Negative expression of LATS1 in glioma WHO grade-4; C. Kaplan–Meier survival analysis of overall survival duration in 103 glioma patients according to LATS1 protein expression. The log-rank test was used to calculate p values. Reduced LATS1 protein expression in glioma We measured the expression levels and subcellular Salubrinal research buy localization of LATS1 protein in archived paraffin-embedded normal brain and glioma samples using immunohistochemical

staining (Figure 1B1-B5). LATS1 protein is primarily localized within the cytoplasm. Furthermore, we observed expression of LATS1 was markedly decreased in glioma samples compared to normal brain tissues (p<0.001) (Table 1). Table 1 The expression of LATS1 protein in Glioma

and normal brain Group   Expression Level of LATS1 Protein(n) P Cases (n) Negative Weak Positive Strong Glioma 103 23 52 20 8   Normal brain 32 1 3 12 16 P<0.001 Relationship between clinicopathologic features and LATS1 expression in glioma patients The relationships between clinicopathologic features and LATS1 expression levels in individuals with glioma were analyzed. We did not find a GSK1904529A mouse significant association of LATS1

expression levels with patient’s age and sex in 103 glioma cases. However, we observed that the expression level of LATS1 was negatively correlated this website with WHO grade (P<0.016) and KPS in glioma patients (Table 2). Table 2 The correlation of LATS1 protein expression with Clinicopathological features in Glioma Clinicopathological features Cases (n) Expression Level of LATS1 Protein(n) P Negative Weak Positive Strong Age ≥55 47 11 22 9 5   < 55 56 12 30 11 3 P = 0.752 Gender Male 60 13 35 7 5   Female 43 10 17 13 3 P = 0.326 WHO grade I 19 1 6 8 4   II 22 3 11 6 2   III 30 7 19 3 1   IV 32 12 16 3 1 P<0.001 KPS             ≥80 53 6 28 13 6   <80 50 14 24 7 2 P = 0.011 Survival analysis To investigate the prognostic value of LATS1 expression for glioma, we assessed the association between levels of LATS1 expression and patients’ survival using Kaplan–Meier analysis with the log-rank test. In 103 glioma cases with prognosis information, we observed that the level of LATS1 protein expression was significantly correlated with the overall survival of glioma patients (Figure 2C). Patients with negative and weak level of LATS1 expression had poorer survival than those with positive and strong level of LATS1 expression (P<0.001). In addition, WHO grade and KPS were also significantly correlated with patients’ survival (P<0.001 and P<0.001 respectively).

Since the https://www.selleckchem.com/products/nepicastat-hydrochloride.html lncRNA PRNCR1 located in 8q24 Vistusertib which was a susceptibility locus to CRC [2–17, 59], we hypothesized that SNPs in this region may have roles in the development of CRC. Our findings confirmed our hypothesis. We found that tag SNPs in the lncRNA PRNCR1 may be a protective

factor against CRC, suggesting that SNPs in lncRNA may be involved in the tumorigenesis of CRC. Although Chung and colleague’s report suggested that the lncRNA PRNCR1 was associated with prostate carcinogenesis and may play a role through the regulation of androgen receptor (AR) transactivation activity [19], no report investigated the relationship between the region 2 of 8q24 and CRC risk. Moreover, there were reports suggested that AR also participated in the pathologic process of CRC through TGFβ pathway [60, 61]. In this study, we found

that the rs1456315 was also associated with clinical features of CRC, which was consistent with the report by Chung et al. [19]. Although we detected the association between SNPs in lncRNA and CRC, there were limitations needed to be mentioned in our study. One is that the follow-up information is blank, which limited our further analysis on the association between SNPs in lncRNA and CRC prognosis. Another is that the study subjects are all ethnic Han Chinese, and the sample size is moderate. Further large-scale studies in different populations, therefore, Transmembrane Transporters modulator still need to be done. Conclusion In conclusion, we found that the variant genotypes of rs13252298 and rs1456315 may contribute to a decreased risk of CRC. Moreover, the rs7007694, rs16901946, and rs1456315 polymorphisms were associated with the tumor size and differentiated status of patients. Association studies with diverse populations and further functional analysis of the variants are needed

to verify our findings. Once our understanding of lncRNAs language is clear, we will be able to classify diseases based on the identified mutations and their effect Acetophenone on lncRNA function. Acknowledgements This work was supported by the special research foundation of doctoral priority to the development of field project (No.20110181130013), the National Natural Science Foundation of China (No. 81302149, 81202387), the Science & Technology Pillar Program of Sichuan Province (14ZC1838, 2013JY0013), Distinguished Young Scientist of Sichuan University (No. 2013SCU04A38), and the Ph.D. Programs Foundation of Ministry of Education of China (No. 20130181120011). Electronic supplementary material Additional file 1: Table S1: Primer sequences and reaction conditions for genotyping the five SNPs. (DOC 36 KB) References 1. Mallardo M, Poltronieri P, D’Urso OF: Non-protein coding rna biomarkers and differential expression in cancers: a review. J Exp Clin Cancer Res 2008, 27:19.PubMedCrossRef 2.

Exacerbating such problems is the fact that many sustainability issues transcend spatial, temporal, sectoral and disciplinary boundaries and thus exceed institutional structures, organizations Savolitinib cost and political mandates. Polk also notes problems in research structures that can hinder the applicability of the results of transdisciplinary research. He cites in particular the lack of an institutional home for practitioners of such research who are not firmly rooted in either the academy or in practice. This, Polk explains,

means that in many cases they risk a lack of legitimacy outside of their immediate sphere of other practitioners. This lack of legitimacy also makes it difficult to capture and utilize project results. He points

to the need for more materials available to scholars that explain these difficulties and how they have been overcome as well as provide examples of how to carry out different types of transdisciplinary research in a variety of substantive areas. There are signs, however, at the international level that channels to decision making may be opening up to transdisciplinary research. The case study by Arico illuminates the way the United Nations and in particular UNESCO is working to achieve the higher Wortmannin research buy level of integration and cross-fertilization of disciplines and to increase stakeholder participation in carrying out its mission to scale up (and to speed up) practical solutions to the sustainable development challenge. Taking this challenge seriously at the behest of its member states, the UNESCO secretariat

is forging ahead with plans to mainstream sustainability science (integrated science for sustainable development) into its various programs. A salient feature of these efforts, and one that is new to the international policy arena, is an overt effort to seek out and include indigenous and local knowledge and to move away from strictly conventional approaches to conducting research and creating new knowledge. In this context, the Arico paper informs us of ways in which the newly launched Future Earth initiative is challenging the conventional 6-phosphogluconolactonase linear model of knowledge production.4 Building on the accomplishments of existing global environmental change programs, the Future Earth initiative was launched shortly before Rio+20, as a new 10-year international research program on global sustainability. This program is designed to mobilize scientists from all disciplines and to strengthen partnerships with stakeholders and selleck screening library policymakers for advancing a global transition toward sustainability. At the heart of this initiative is the idea of co-design of research through a higher level of interaction between stakeholders and scientists.

Pam binds to EPS in the check details extracellular matrix and modifies cell attachment To investigate the localization of Pam in P. luminescens TT01 cells, sections of bacterial colonies were observed under transmission electron microscopy (TEM) revealing large amounts of exopolysaccharide (EPS)-like matrix filling the spaces between cells (Fig. 4A). We used immunogold localization of Pam in these sections and found that the protein is associated with this extracellular material that is distributed surrounding the cells (Fig. 4B). In TT01pam the EPS-like material was still present but we did not see specific binding of the antibody (Fig. 4C), suggesting that although Pam binds to the extracellular matrix, it does not

significantly alter its production or general structure. Furthermore, Western-blot analysis using the anti-Pam antibody revealed that Pam could be detected in crude EPS preparations (Fig 4D), confirming that from all the extracellular matrix components Pam binds at least to EPS. Our studies revealed that EPS-bound Pam can be released by the action of SDS and salt (KCl) but not by mechanical disruption (vortex) (data not shown). Eltanexor nmr Figure 4 Pam localization on bacterial cells. (A) Micrograph www.selleckchem.com/products/azd1080.html of a cross-section from a P. luminescens TT01 colony observed by TEM. Note the presence of an extensively folded extracellular matrix (black arrow) between the bacterial cells (indicated with

P). (B) Immunolocalization of Pam using the anti-Pam antibody and a conjugated-gold secondary antibody. Gold particles extensively decorate the fibrillar EPS-like matrix (black arrow). (C) The TT01pam strain shows no anti-Pam antibody signal but the fibrillar

matrix is still present. Scale bars are 0.2 μm. (D) Western blot confirming the presence of Pam in preparations of crude EPS. Lane 1: crude EPS extracted from TT01rif, lane 2: EPS from TT01pam and lane 3: purified recombinant Pam. As Pam binds to EPS and EPS has been shown to be important in biofilm formation [11], we investigated the possibility that Pam influences the different stages of biofilm formation. Pellicle assays and biofilm growth in microscopy chambers did not show differences in mature biofilm formation between TT01rif and TT01pam (data not shown). To analyze the influence of Pam on the early steps of biofilm formation, namely Baf-A1 in vitro initial attachment, we looked at attachment of the two strains to glass coverslips when cultured ex vitro in hemolymph plasma. As shown in Figure 5, the parental TT01rif cells attached in greater numbers than TT01pam to the glass surface in hemolymph, but not in LB medium or Schneiders insect growth medium (data not shown). Importantly, we were also able to detect Pam in cell and supernatant fractions in bacteria grown in hemolymph plasma at 8 hours. Figure 5 Comparison of bacterial attachment to surfaces in presence of insect hemolymph by fluorescence microscopy between TTO1rif and the pam mutant.

Testing a larger collection of strains from diverse origins could address this question. Diverse methods have been proposed for the molecular typing of bacteria in the genus Ochrobactrum. ITS1 sequencing and rep-PCR have been successfully used to assess the level of microdiversity in the genus as well as to cluster the strains according to the species [12, 13]. However, within the species O. anthropi there was no correlation between

rep- or ITS1-based clusters and origin of the strains. In the collection selleck chemical tested, MLST data and multi-locus-based phylogeny provided this website evidence of a clonal complex associated to human beings. To strengthen this evidence, the question of the representativeness of the human strains included in the MLST analysis should be addressed. Most clinical strains originated from France (n = 34) but they have been isolated in diverse regions and at different times from 1998 to 2007. We also included 9 geographically unrelated clinical strains isolated in

Scandinavia, United Kingdom or Louisiana (USA) from 1971 to 1995. Seven of them belonged to the major complex MSCC4/eBCC4 beside most of the French clinical isolates. This indicated that MSCC4/eBCC4 could be considered as selleck a human-adapted subpopulation rather than a geographic subpopulation. The mean genetic diversity calculated from the seven loci showed no significant differences between clinical isolates and isolates from all other various origins. This is also the case for the number of STs per strain. The genetic

diversity of the clinical population was confirmed at the genomic level since all the clinical strains displayed different pulsotypes indicating that they were epidemiologically unrelated. Therefore, epidemiological, genetic and genomic data exclude a bias in strain sampling and enhance the robustness of the human-associated subpopulation described herein. PFGE typing appeared highly discriminative in the species O. anthropi since only 2 strains originating from the same environmental sample displayed Selleck Sirolimus the same pulsotype. None of the isolates originating from one hospital displayed the same pulsotype. This wide genomotype diversity observed here confirmed previous data showing the genomic plasticity of O. anthropi [28]. Genomic rearrangements in plastic genomes are considered as rapid evolution mechanisms, named micro-evolution with respect to the time-scale, that could be involved in rapid adaptation processes to a particular niche [42]. Restriction fragment length polymorphism in PFGE detected genomic modifications such as rearrangements and horizontal genetic transfer events rather than single nucleotide polymorphisms [43]. The higher discriminative power of PFGE suggested that large rearrangements occurred at higher rates than intragenic point mutations in housekeeping genes in O. anthropi.

Thus investigation of detailed Quisinostat supplier vaccine induced cell-mediated response after immunization may help to understand the underlying mechanism of different formulations that can correlate with the observed protection. Next, we evaluated the Th1 and Th2 cytokine responses in differently adjuvanted mice. Splenocytes from immunized mice were isolated 10 days after immunization and, IFN-γ and IL-4 levels were measured in vitro following restimulation with LAg. LAg in different adjuvant vaccinated groups produced substantial amounts of IFN-γ compared to controls (Figure 4A; P < 0.001). Interestingly, the most pronounced increase in IFN-γ level was observed in MPL-TDM+ LAg vaccinated

groups in comparison to other groups (P < 0.001). Mice immunized with BCG+LAg

secreted lower amount of IFN-γ compared with the liposomal LAg immunized group (P < 0.05). Mice receiving BCG+LAg and liposomal LAg immunization showed significant increase in IL-4 production compared to controls (Figure 4B, P < 0.001). However, elicitation of significantly higher IL-4 response was observed in liposomal LAg vaccinated mice compared to BCG+LAg immunized groups selleck (P < 0.01). In contrast to the robust IFN-γ responses observed with MPL-TDM+LAg vaccine, IL-4 level was significantly lower from other vaccinated groups (P < 0.01). Thus, MPL-TDM+LAg triggered highest IFN-γ but lowest IL-4 indicating an exclusive

Th1 cell-mediated immune response. BCG+LAg and liposomal LAg generated a mixed Th1/Th2 response as evident from significant production of both IFN-γ and IL-4 post-immunization groups. But compared to the Th1/Th2 response generated by liposomal LAg, the cytokine levels were lower for BCG+LAg immunized groups. Figure 4 IFN-γ and IL-4 responses in differently adjuvanted LAg vaccinated mice . Mice were immunized three times at 2-week intervals. Ten days after last immunization spleens were collected from mice and restimulated in vitro with LAg (10 μg/ml). After 72 h supernatants were collected and concentrations of released IFN-γ (A) and IL-4 (B) levels were determined by ELISA. Each sample else was click here examined in duplicate. Each bar represents the mean ± SE for five individual mice per group. The results are those from one experiment representative of two performed. Asterisks over each bar indicate significant differences in comparison to control groups. Asterisks over line indicate significant differences between groups. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Discussion Despite the current knowledge of immunology and pathology related to the parasite Leishmania, till now, a desirable vaccine for humans has not been successfully developed. The main goal of vaccination is the induction of a protective immune response against the pathogen.

However, its activity depends on environmental stimuli (e.g., cyclic AMP levels, temperature, heat shock, osmolarity, membrane

biosynthesis, and H-NS protein [8]), cell division, flagella formation, and motility [9–11]. A number of Gram-negative pathogenic bacteria have evolved a specialized type III protein secretion system to deliver effector virulence proteins into host cells [12, 13]. There #AG-881 mw randurls[1|1|,|CHEM1|]# are two types of type III secretion systems: the translocation-associated type III secretion system (T3aSS) and the bacterial flagellum type III secretion system (T3bSS). The various bacterial type III secretion systems characterized thus far all have Sec independence, ATPase dependence, presence of a hollow filamentous organelle that extends from the outer membrane, a cell-envelope-spanning secretion channel, and nine conserved proteins [14]. The bacterial flagellum type III secretion system also serves as the bacterial flagellum (a biological nanomachine with an ion-powered rotary motor). For the flagellum, the T3bSS apparatus functions to secrete components including the rod, hook, and filament subunits for extracellular assembly. The core of the flagellum is hollow, and secreted subunits polymerize at the growing end of the flagellum. A cap at the tip of the flagellum ensures efficient polymerization check details of secreted subunit proteins [15, 16]. This secretion

apparatus is just one mechanism utilized by Gram-negative plant and animal pathogens for the secretion and translocation of virulence determinants into susceptible eukaryotic cells [17]. In Salmonella typhimurium, the expression of class 1 genes (i.e., flhD and flhC) activates expression of N-acetylglucosamine-1-phosphate transferase genes required for flagella assembly and regulates expression class 2 genes (e.g., fliAZY and flhBAE), which in turn regulates expression of class 3 genes encoding flagellar structural proteins (e.g., fliC, flgMN, and MotAB) [18].

In Xenorhabdus nematophila, it was shown that the EnvZ-OmpR-FlhDC-FliA regulatory network coordinately controls flagella synthesis as well as exoenzyme and antibiotic production [8]. In this paper, we describe the transcriptional regulation of fliC and flhA expression by flhD/C and also show that flhD/C has an effect on extracellular secretion of the Carocin S1 protein, but not on Carocin S1 gene expression. Our results indicate that the type III secretion system of Pectobacterium carotovorum subsp. carotovorum has a new secretory function. Methods Bacterial strains, plasmids, media, and growth conditions The strains and plasmids used are shown in Table 1. Pectobacterium carotovorum subsp. carotovorum strains were propagated at 28°C in 1.4% nutrient agar (NA) or with shaking in Luria-Bertani (LB) medium with NaCl (5 g/L). E. coli strains were propagated at 37°C in LB medium with shaking. Rifampicin, kanamycin, and ampicillin (all at 50 mg/L) were added to either medium when necessary.

PCR products were purified with QIAquick PCR Purification Kit (Qiagen) and sequenced with primers fD1, rP2 and R1087 (5’-CTCGTTGCGGCACTTAACCC-3’), gyrA-5F, and gyrB-F1, respectively. Sequencing was done in the Department of Entomology at the Max Planck Institute for Chemical Ecology (Jena, Germany) or commercially by SEQLAB Sequence Laboratories (Göttingen, Germany).

Bacterial sequences were deposited in the GenBank database under following accession numbers: KM035545 – KM035652 (16S rRNA genes), KM035653 – KM035673 (gyrA genes) and KM035674 – KM035755 (gyrB genes). Diversity of bacterial strains in individual beewolf antennae Bacterial micro-colonies see more were isolated from individual antennae of two different Philanthus multimaculatus and one Philanthus psyche

female with serial dilution in Selleckchem Omipalisib 24-well plates with liquid medium Vactosertib price as described above. Individual micro-colonies were carefully transferred by pipette into 96-well PCR plates with 100 μl PCR lysis solution A without proteinase K (67 mM Tris–HCl (pH 8.8); 16.6 mM (NH4)2SO4; 6.7 mM MgCl2; 6.7 μM EDTA (pH 8.0); 1.7 μM SDS; 5 mM β-mercaptoethanol) [40]; samples were heated at 95°C for 5 min to destroy bacterial cells. Afterwards, gyrB gene fragments were amplified, purified and sequenced as described above. Obtained sequences were aligned and manually curated using Geneious software version 6.0.5 (Biomatters Ltd., http://​www.​geneious.​com/​). until Phylogenetic analysis 16S rRNA, gyrA and gyrB gene sequences of isolated symbionts were aligned with those obtained from field-collected beewolves

as well as representative outgroup sequences of free-living Streptomyces and other actinomycete strains (Additional file 4: Table S4). Alignments of individual genes were concatenated for phylogenetic analyses. Approximately-maximum-likelihood trees were reconstructed with FastTree 2.1 using the GTR model, with local support values estimated by the Shimodaira-Hasegawa test based on 1,000 resamplings without re-optimizing the branch lengths for the resampled alignments [41]. Bayesian inferences were run with MrBayes 3.1.2 [42–44], with the different genes defined as separate partitions in the concatenated alignment. The searches were conducted under the GTR + I + G model, with 4,000,000 generations per analysis. Trees were sampled every 1,000 generations. We confirmed that the standard deviation of split frequencies was consistently lower than 0.01, and a “burnin” of 25% was used, i.e. the first 25% of the sampled trees were discarded. We computed a 50% majority rule consensus tree with posterior probability values for every node.

This is typically the profile recovered from the SGI1, and therefore was designated as IP-SGI1 (Figure 2B and Additional file2).

Sequence determination for three isolates showed that the 1,000 bp cassette contained aadA2 and that the 1,200 bp cassette coded for pse-1, which are the most commonly this website found integrons in the SGI1. All the isolates were positive for the amplification of pse-1and aadA2 using primers specific for these genes (Figure 2B and Additional file3). To confirm the insertion of the complete SGI1 in the chromosome, we performed PCR assays to amplify the left and right junctions. All the isolates (n = 19) harbouring the IP-SGI amplified the left junction, the right junction, and were positive for the amplification of the cryptic retronphage

on the right junction [see Additional file2]. Isolates harbouring other integrons did not amplify any of the junctions of the SGI1. To further characterize the SGI1, we amplified RAD001 the tetG and floR genes that are in between the two integrons. Only the isolates harbouring the IP-SGI1 produced strong amplification products with tetG, and all were positive for floR; however, other chloramfenicol selleck products resistant isolates also amplified floR. All the cmy-2 positive isolates (n = 36) were positive for floR, which is in agreement with the report by Doublet et al. (2004) that both resistances are often found in the Farnesyltransferase same plasmid [11, 48]. Thus, most of the floR positive isolates harboured SGI1 or pCMY-2, however, other chloramfenicol resistant isolates were positive for floR. Some of the

isolates harbouring IP-2 showed weak amplification bands with tetG or floR primers, probably due to the presence of related but divergent genes conferring resistance to tetracycline and chloramfenicol [see Additional file2]. Two significant associations among integrons and the other molecular markers are worthy of mention. First, all IP-1 were carried by ST213 isolates (p = 0.001, OR = 211), either cmy-2 positive or negative. Second, all the isolates with SGI1 were ST19 and carried pSTV (p = 0.001, OR = 119), the only exception was one isolate that did not carry pSTV (yuhs00–141; Figure 4 and Additional file2). To determine the location of the integrons, we performed Southern hybridization experiments using fragments of the intI1 and aadA2 genes as probes on the plasmid profiles of eight representative isolates. Three of the five isolates harboring IP-1 hybridized with a plasmid of about 100 kb, the remaining two IP-1 isolates hybridized with a plasmid of about 150 kb. The isolate harboring IP-2 hybridized with a plasmid of about 150 kb, IP-3 with a plasmid of about 35 kb, and IP-4 with a plasmid of about 100 kb. Detection of intI1 and qacEΔ1 To further characterize the 5′ and 3′ CSs of integrons we amplified intI1 and qacEΔ1 (Figure 2A).

It plays essential roles in promoting cell proliferation [8–11]. Our previous studies have shown that HSP70 could interact with C23 and inhibiting H2O2-induced cleavage and degradation of C23, thereby inhibiting reactive oxygen species-induced cell apoptosis [12]. There Lorlatinib were two ways for radiotherapy to destruct tumor cells: (1) X-ray directly broke the DNA of the cancer cells into fragmentations, leading to cell apoptosis; (2) X-ray released free radicals from other components (e.g. H2O) in the cells thereby to attack tumor cells. Theoretically, radiotherapy could result in cleavage and degradation of C23 and sequentially kill the tumors. In the present study, we determined whether reduction

of HSP70 expression could enhance radiosensitivity

of LSCC by increasing C23 cleavage and degradation. Materials and methods Tissue Microarray High-quality tissue microarray (TMA) was constructed with fifty tumor samples including CHIR98014 price different stages of LSCC. The clinicopathologic features of the participants included in this analysis were presented in Table 1. Briefly, serial 5-μm sections were cut from each of the donor blocks. One of Ricolinostat purchase these sections was stained with hematoxylin and eosin staining (H&E) to mark morphologically representative areas of the tumor. Two areas in each case were targeted. Tissue cylinders with a diameter of 0.6 mm were punched from the two targeted areas in each donor Selleckchem ZD1839 block and deposited into a 14 × 7+2 (100 cores) TMA block, which

contained 50 cores of tumor tissues. At last we gained 80 slides of high-quality TMA. Immunostaining for HSP70 protein was performed by using TMAs. Table 1 Clinicopathologic characteristics of participants of TMA Clinicopathologic characteristics of participants of TMA Male 45 Female 5 Average Age 61.3 ± 4.2 Stage I, II 21 Stage III, IV 29 RNA oligos According to the design principle of oligodexynucleotide (ODN) probes described by Myers KJ and Branch AD [13, 14], three antisense-ODNs (ASODNs) were designed artificially against the HSP70 mRNA complete sequence (GeneBank NO.BC002453) from http://​www.​ncbi.​nlm.​nih.​gov/​. Three ASODNs were synthesized with phosphorothioate modification by Bioasia Co. Ltd. (Shanghai, China). After screening an effective ASODN, AS-1(5′-X TGTTTTCTTGGCCAT -3′), which complemented to the first 20 coding sequences of HSP70 mRNA, random oligos (5′-X GATTATCGTGTTGTTACT -3′) were used as negative controls against AS-1, X represents green fluorescent marker. Animals and treatment BALB/c female mice (18-22 g, 4-6 weeks) were obtained from Laboratory Animal Centre, Xiangya School of Medicine, Central South University (changsha, China). The animals were housed for 1 week prior to experiment. The animal experiments were undertaken within the guidelines of regulations for the use of experimental animals of Central South University.