Three loci have already been described VS-4718 by Radtke et al. in a contemporary

study but were amplified here with other primers [32] (Table 2). For the SAG7 locus, no amplification was observed with primers directly flanking the TR for 14% (26/189) of the strains. A second primer pair targeting larger consensual flanking regions was designed to confirm the absence of the locus. PCR was performed in a final volume of 25 μl containing 10 ng DNA, 1 × PCR Reaction Buffer, 2 mM MgCl2 (Applied Biosystems), 5% DMSO (dimethyl sulfoxide), 1 unit of Taq DNA polymerase (Applied Biosystems), 200 μM of each dNTP and 0.5 μM of each flanking primer (Eurogentec, Belgium). Amplification was performed in a 2720 Thermal Cycler (Applied Biosystems) under the following conditions: initial denaturation for 5 min at 94°C, followed by 30 cycles of denaturation for 30 s at 94°C, annealing for 30 s at 50°C and elongation for 60 s at 72°C plus a final elongation step for 7 min at 72°C. We separated 10 μl of PCR product by electrophoresis in a 2% agarose gel (Eurogentec, Belgium), which was also loaded with a 100 bp DNA size ladder (New England BioLabs). Electrophoresis was performed in 20 cm-long

gels, in 1× TBE buffer (89 mM Tris-Borate, 2.5 mM EDTA) containing 1 μg/ml ethidium CP673451 cell line bromide run at 10 V/cm. In each run, at least one lane was loaded with PCR product from one of the reference strains, NEM316, A909 or 2603 V/R. The gels were photographed under ultraviolet illumination, with Vision-Capt® Software

(Vilber-Lourmat, Marne la Vallée, France). The number of repeats for each VNTR was deduced from amplicon size, by comparison with the reference strain, for which the number of repeats was known. The allele number corresponded to the number of repeats. For the SAG7 locus, the lack of a VNTR was revealed by the absence of amplification with the first Loperamide primer pair and the amplification of a fragment of the expected size with the second primer pair, which targeted larger consensual flanking regions. In this case, an allele number of 0 was given. For the SAG21 locus, a 117 bp PCR product was obtained, demonstrating deletion of the inserted sequence and, thus, the absence of a VNTR. An allele number of 0 was also assigned in this case. The MLVA genotype of a strain was expressed as its allelic profile, corresponding to the number of repeats at each VNTR, listed in the order SAG2, SAG3, SAG4, SAG7, SAG21, SAG22.

For example, the electrode materials can influence the electronic coupling between electrodes and molecules, such as the interaction of electrode-anchoring group and the alignment of the energy level of electrode-molecule [8, 9]. Typically, most of the conductance measurements of single-molecule junctions were performed by using Au as electrode for its chemically inert property [10]. However, it is also important to study the non-Au electrodes to fully understand the charge transport through single-molecule junctions. We pay attention to the Ag electrodes for the following reasons: Ag has strong optical enhancement property and high catalytic activity [10–12]. It

has a similar electronic structure with Au and Cu and is easy for comparison among them. Single-molecule conductance can be measured by scanning tunneling microscopy (STM) break junction (STM-BJ), mechanically controllable break junction DihydrotestosteroneDHT research buy (MCBJ), STM trapping and conducting atomic force microscopy, and so on [13–21].

Though lots of works have been done on the electron transport of single-molecule junctions by using the above methods, there is limited investigation on single-molecule junctions with non-Au electrodes [10, 22]. We have developed an electrochemical jump-to-contact scanning tunneling microscopy break junction approach (ECSTM-BJ) [23]. By using this approach, single-molecule junctions with carboxylic acid binding to different metallic electrodes were systematically investigated [9, 24]. Since the pyridyl group also ��-Nicotinamide mw has received much attention [15, 17, 25–27], we recently extended this approach to the conductance measurement of pyridyl-based molecules binding to Cu electrode, which shows that the single-molecule conductance with pyridyl-Cu contacts

is smaller than that with pyridyl-Au contacts [28]. In this work, we focus on the single-molecule junctions with pyridyl group (Figure 1a) binding to Ag contacts by ECSTM-BJ. Especially, the influence of the electrochemical potential on the Fermi level of electrode is discussed. Figure 1 Molecular structure and schematic diagram of ECSTM-BJ. (a) Molecular structures of 4,4′-bipyridine (BPY), 1,2-di-(pyridin-4-yl)ethene Smoothened (BPY-EE), and 1,2-di(pyridin-4-yl)ethane (BPY-EA), and (b) schematic diagram of Ag-molecule-Ag junctions formed by the ECSTM-BJ. Methods Au(111) was used as substrate, and mechanically cut Pt-Ir (Φ = 0.25 mm) wires were used as the tips. The latter was insulated by the thermosetting polyethylene glue to reduce the leakage current of the electrochemical reaction. Ag and Pt wire were used as the reference and counter electrodes, respectively. 1,2-Di(pyridin-4-yl)ethene (BPY-EE) and 1,2-di(pyridin-4-yl)ethane (BPY-EA) were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA), while 4,4′-bipyridine (BPY) and Ag2SO4 (99.999%) were purchased from Alfa Aesar (Ward Hill, MA, USA). H2SO4 was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).

A) 2 days post-infection (dpi). B) 4 dpi. C) 9 dpi. Top panel shows mean mapped reads and count distribution along DENV2 genome for a representative library at each time point. Bottom panel shows mean viRNA distribution by sRNA size group. Blue and red bars indicate sense P505-15 mw and anti-sense viRNAs, respectively. Host sRNA Profiles To identify host factors that

are differentially regulated by SRRPs during DENV2 infection, we asked whether sRNA profiles mapping to host RNAs change in DENV2-infected mosquitoes compared to un-infected controls. sRNA profiles were categorized by the target RNA to which they mapped, as well as by sRNA size group. Changes to ncRNAs were also measured, because NVP-BSK805 recent evidence suggests that they are regulated by RNAi pathway activity [28, 32]. For this line of inquiry, we did not distinguish between 20-23 nt siRNAs, endo-siRNAs, or microRNAs. We reason that enriched sRNA profiles for a given target represent the product of enhanced target cleavage, in the absence of concomitant transcriptional repression or mRNA decay [28, 33]. Conversely, depleted sRNA profiles among the DENV2-infected pools would be indicative of fewer degraded mRNAs. We defined a single sRNA profile as the number of reads mapped to a single target transcript. The presence of sRNAs aligning to a given transcript would be expected to change

sporadically across the three biological replicates if they arose through MYO10 non-specific decay events. Moreover, non-specific decay events should produce sRNAs across all size groups in similar frequency. Therefore, we expect that sRNA levels with statistically significant enrichment or depletion represent altered RNAi pathway activity. The RNA-seq

program edgeR was used determine the significance of sRNA profile enrichment or depletion for sense and anti-sense sRNAs across all three replicates for each timepoint [34]. Sense strand reads were more abundant than anti-sense reads. All target transcripts are categorized by read orientation in Additional File 2. A cut-off value of 0.05 False Discovery Rate (FDR) was used to determine whether changes were statistically significant [35]. sRNA populations mapping to mRNAs and ncRNAs were grouped into functionally similar categories. Figure 3 shows functional categories for which sRNA profiles were modulated over the course of infection. At 2 dpi, 555 unique targets showed enriched sRNA profiles compared to controls, whereas at 4 dpi, only 67 targets had significantly enriched sRNA profiles (Figure 3A and Additional File 2). Under-represented sRNA profiles were much less abundant; 43 unique targets showed depleted sRNAs in DENV2-infected mosquitoes at 2 dpi, and 44 targets showed depleted sRNAs at 4 dpi (Figure 3B). Very few differentially abundant profiles were observed at 9 dpi, therefore they were excluded from further analysis (Additional File 2).

Electrophoretic mobility shift assay DNA fragments used for the electrophoretic mobility shift assay (EMSA) were PCR amplified using Cy5-labeled primers to perform HSP inhibitor a non-radioactive EMSA. DNA fragments used were the upstream region of acrD (246 bp), and as controls, the upstream regions of acrAB (205 bp) and tolC (291 bp). Approximately 0.16 pmol of Cy5-labeled DNA was mixed with increasing concentrations of His-tagged BaeR protein in a binding buffer reaction (50 mM

Tris–HCl, pH 7.5; 1 mM DTT; 500 mM MgCl2; 100 mM EDTA; 10 mM NaCl; 5% glycerol). To decrease unspecific binding, 500 ng competitor DNA (Salmon sperm DNA, AppliChem) was added to the reaction. Incubation was done at room temperature for 30 min. The total reaction was run on a native 4% polyacrylamide gel in 0.5x Tris-borate-EDTA (TBE)

buffer at constant 25 mA. After electrophoresis, fluorescence signals of the labeled DNA were visualized using a FLA-3000 phosphorimager (Raytest, Straubenhardt, Germany). Statistical analysis Statistical analysis was performed using R [56]. Differences between two groups were determined by a two-sided t-test with equal variances and were considered significant at P < 0.05. When necessary the standard deviation is presented in the graph when the average of several values was applied. Acknowledgments This study was supported by Jacobs University Bremen and by the MOLIFE Research Center, Selonsertib research buy Jacobs University Bremen. Electronic supplementary material Additional Flavopiridol (Alvocidib) file 1: Phylogenetic tree of AcrD. Description: The tree was calculated based on AcrD from Erwinia amylovora Ea1189 (black arrow) and its homologues from other members of the Enterobacteriaceae family, including Erwinia pyrifoliae (95% identity), E. tasmaniensis (93% identity), E. billingiae (83% identity), Pantoea agglomerans (82% identity), P. ananatis (79% identity), Enterobacter cloacae (79% identity), Salmonella enterica (79% identity), Citrobacter koseri (79%), Klebsiella pneumoniae (79% identity), Escherichia coli (78% identity) and Shigella flexneri (78% identity). The dendrogram was generated based on percentage of identity

between the sequences using the neighbor joining algorithm implemented in Jalview [25–28]. (TIFF 6 MB) Additional file 2: Sequence alignment of AcrD from Erwinia amylovora Ea1189 and Escherichia coli K-12. Description: The alignment is based on the amino acid sequences of AcrD using ClustalW for analysis and Jalview for data presentation. AcrD of Ea1189 is 79% identical and 89% similar to AcrD of E. coli K-12. Identical amino acid residues are shown in blue. Yellow bars show a quantitative measurement of conserved physico-chemical properties where the highest score shows amino acids of the same physico-chemical class [26–28]. Black bars indicate predicted transmembrane-spanning helices of AcrD from E. amylovora[29]. (TIFF 5 MB) Additional file 3: Modified view of the genomic organization of the acrD locus.

Splenic infarction following cocaine use is rare but has been described, particularly in patients with sickle hemoglobinopathies [8]. It is plausible that cocaine-associated splenic hematoma or rupture results from transient vasospasm with subsequent bleeding into the infarcted area. Secondary infection of the infarcted spleen with resultant sepsis and death has also been LEE011 order detailed [9]. While the use of cocaine causing hematoma of the spleen has been described [10], this case is the first report of a case that details hemoperitoneum caused by ASR following cocaine use. Although uncommon, the potential for death due to splenic rupture warrants awareness and highlights the importance of

a social history in patients presenting with acute abdominal pain. Consent Written informed

consent was obtained from the patient for publication of this Case report and any accompanying SN-38 in vivo images. A copy of the written consent is available for review by the Editor-in-Chief of this journal. Acknowledgements We would like to thank Dr. Stephan Anderson for providing the representative images and captions. References 1. Renzulli P, Hostettler A, Schoepfer AM, Gloor B, Candinas D: Systematic review of atraumatic splenic rupture. Br J Surg 2009,96(10):1114–1121.PubMedCrossRef 2. Wehbe E, Raffi S, Osborne D: Spontaneous splenic rupture precipitated by cough: a case report and a review of the literature. Scand J Gastroenterol 2008,43(5):634–637.PubMedCrossRef 3. Debnath D, Valerio D: Atraumatic rupture of the spleen in adults. J R Coll Surg Edinb 2002, 47:437–445.PubMed 4. Amonkar SJ, Kumar EN: Spontaneous rupture of the spleen:

three case reports and causative processes for the radiologist to consider. Br J Radiol 2009, 82:e111-e113.PubMedCrossRef 5. Tinkoff G, Esposito TJ, Reed J, Kilgo P, Fildes J, Pasquale M, Meredith JW: American Association for the Surgery of Trauma Organ Injury Progesterone Scale I: spleen, liver, and kidney, validation based on the National Trauma Data Bank. J Am Coll Surg 2008,207(5):646.PubMedCrossRef 6. Kaufman MJ, Siegel AJ, Mendelson JH, Rose SL, Kukes TJ, Sholar MB: Cocaine administration induces human splenic constriction and altered hematologic parameters. J Appl Physiol 1998,85(5):1877–1883.PubMed 7. Bellows CF, Raafat AM: The surgical abdomen associated with cocaine abuse. J Emerg Med 2002,23(4):383–386.PubMedCrossRef 8. Vaghjimal A: Splenic infarction related to cocaine use. Postgrad Med J 1996,72(854):768.PubMedCrossRef 9. Dettmeyer R, Schlamann M, Madea B: Cocaine-associated abscesses with lethal sepsis after splenic infarction in an 17-year-old woman. Forensic Sci Int 2004,140(1):21–23.PubMedCrossRef 10. Homler HJ: Nontraumatic splenic hematoma related to cocaine abuse. West J Med 1995,163(2):160–162.PubMed Competing interests The authors declare that they have no competing interests.

There is a discontinuous narrow coastal terrace, on which most development has occurred (Fig. 8b), and a fringing reef with a number of reef-gap beaches. In addition to coastal hazards, rockfall and landslides are a threat to development on the selleck chemicals coastal terrace beneath

steep slopes. Fig. 8 a Reef-fronted beach with outcrop of granite and beachrock (foreground), east coast of high island of Mahé, Seychelles (photo DLF 2005). Note hotel overhanging seawall and beach. b Development on coastal terrace, Baie de la Mouche, west coast of Mahé, where natural berm has been removed for road construction: tsunami damage occurred here in 2004 (photo DLF 2005) Coastal hazards on small islands The MRT67307 research buy nature of the hazards, exposure and vulnerability—thus the most relevant adaptation measures—vary between island types in relation to elevation, but also to size, topography, bathymetry, lithology, reef morphology and ecological integrity, as well as human factors such

as shore protection, or location and design of critical infrastructure and other property. The geographic region is important as it determines ocean climate (e.g., temperature and coral growth rate), storm climatology (including wind and wave patterns), and the regional trend of sea-level rise. Islands within ± 5° latitude about the equator are generally free of tropical cyclones, but occasional storm incursions, exceptional ADP ribosylation factor winds, or impacts of far-travelled swell from mid-latitude storms can cause significant damage, the effects of which are also influenced by sea-level variability resulting from El Niño-southern oscillation (ENSO) or other large-scale climate cycles. At tropical to mid-latitudes >5° (north or south),

tropical cyclones are a major recurring threat (Hay and Mimura 2010). In addition to climate effects, geophysical hazards such as volcanic eruptions, landslides, earthquakes and tsunami require attention and may pose equal or greater risks to island communities. Apart from catastrophic events, coastal stability is a function of wave energy, erodibility, and sediment supply, which may depend on reef health and the production of biogenic sand (Kench and Cowell 2001; Perry et al. 2008, 2011). Reefs represent not only a source of sediment, but play a major protective role, absorbing much of the deep-water wave energy. There is cause for concern about the mid-term fate of coral reefs (e.g., Hoegh-Guldberg et al. 2007), but recent work has shown that the coralline algae forming the resistant rims of some reefs may be more resistant to acidification than previously thought (Nash et al. 2013). In some places, exposure is mitigated and resistance to erosion increased where mangroves are present along the shore. Removal of mangroves can often be identified as a source of erosion problems in coastal communities (Mimura and Nunn 1998; Solomon and Forbes 1999).

Results Contractile response of vascular ring to NA Vascular dysfunction is related to increased vasoconstriction and

weakened diastolic function. Therefore, we are interested in determining whether there is any change in the vascular function by detecting the vascular reactivity of aortic rings S3I-201 to a physiological modulator, noradrenaline (NA). Cumulatively added NA (10-10-10-5M) caused concentration-dependent contractile responses in isolated aortic rings. We found that there was no significant difference between the SE and the CS group, while the ES group significantly increased the vasoconstrictive response to NA (P<0.01), LBPs treatment decreased the vasoconstrictive effect ( P< 0.01) (Figure 1). Furthermore, the contractile responsiveness to NA of the SE group was significantly lower than that of the ES (P<0.01) and ES-LBP (P<0.01) groups (Figure 1). Figure 1 Contractile response of vascular ring to NA. Dose-dependence of NA on contraction of the thoracic aorta rings separated from rats in CS SE, ES and ES-LBP groups. The contraction induced by 60

mM KCl was taken as 100%. Data are expressed as mean ± SD (n=10). # P<0.01 vs CS; ※ P<0.01vs SE; JQ1 △ P<0.01 vs ES. Effects of LBPs on body weight and exhaustive exercise time in rats After four weeks of swimming exercise, no significant difference was observed in body weight in either group (Table 2). However, as shown in Figure 2, LBPs prolonged the swimming time of rats compared with the ES group ( P ROS1 < 0.05), which was 77.07% higher. Table 2 Effects of LBP on body weight in rats Group Before experiment One week Two week Three week Four week CS 191.67±26.90 204.83±13.43 264.08±12.31 304.44±9.97 346.58±15.55 SE 187.5±4.74 209.53±6.15 258.43±9.88 309.35±19.11 340.5±22.31 ES 191.2±10.77 210.67±10.91 263.5±14.05 304.58±17.12 329.13±15.06 ES-LBP 198.2±9.66 215.14±7.22 267.70±6.96 312.08±10.14 344.33±14.91

Effects of LBPs on body weight in rats. The values are expressed as mean ± SD (n=10). Figure 2 Effects of LBPs on exhaustive exercise time in the rats. LBPs supplementation significantly increased the time to fatigue compared to that of the ES. Data are mean ± SD (n =10). △ P < 0.01 vs ES. Effects of LBPs on biochemical parameters after exhaustive exercise It is well known that SOD can inhibit the oxidation of oxyamine by the xanthine–xanthine oxidase system. Therefore we evaluated the plasmic level of SOD. As shown in Figure 3a, the SOD level in the ES-LBP, SE groups significantly increased compared with that in the CS group (P<0.05 and P<0.01 respectively). However, the plasmic SOD level of exhaustive swimming rats was significantly lower than that of the ES-LBP and SE rats (P< 0.01). The results demonstrated that LBPs were able to increase antioxidant enzyme activities to attenuate the oxidative stress induced by exhaustive exercise. Figure 3 Effects of LBPs supplement and exhaustive exercise on SOD (a), MDA (b), NO (c) and HSP70 (d) expression in the rats.

4%) in a population of 125 B. bassiana isolates [25]. The number of introns found in the 57 isolates was in agreement with the 199 introns detected in 125 B. bassiana isolates by Wang et al. [25]; the 44 introns detected in 26 M. anisopliae isolates by Márquez et al. [31], and the 69 introns found in 28 representative

members of the genus Cordyceps by Nikoh and Fukatsu [26]. However, only four intron insertion patterns were present in our B. bassiana collection while greater variability was found in other studies: 13, 7 and 9 insertion patterns within 125 B. bassiana [25], 26 M. anisopliae [31] and 47 B. brongniartii Thiazovivin ic50 [23] isolates, respectively. The MP tree based on intron sequences shows that they were distributed in four large groups, with bootstrap values of 100%, corresponding to four insertion positions (Figure 1). As could be expected [25, 28], the introns inserted at the same site always belonged to the same subgroup: IC1 at positions 2 and 4, and IE at position 1. Although the ARRY-438162 chemical structure origin and transmission mechanisms of group I introns have generated controversy [26], this distribution of sequences is in agreement with previously reported observations [25] and means that introns inserted at the same position have a monophyletic origin and are transmitted vertically. In subsequent events intron speciation

and diversification take place as occurs at position 4, where B. bassiana introns are separated from Metarhizium and Cordyceps introns, and two B. bassiana IC1 sequence sizes were located in two different sub-clades, supported by high bootstrap values. Rehner and Buckley’s study [8] based on EF1-α and ITS phylogenies has revealed that i) six clades can be resolved within Beauveria (A-F) and, excepting those corresponding to B. bassiana (A and C), they are closely

to species previously described on the basis of their morphology, and ii) B. bassiana s.s. (A) was determined almost entirely from nucleotide variation at EF1-α. Further phylogenetic studies carried out with nuclear and/or mitochondrial DNA regions of B. bassiana from all continents have served to resolve BCKDHB lineage diversity within this species [7, 12, 18, 21]. Since phylogenetic species by continent and in the order of their discovery have been designated previously [7], we followed this nomenclature to refer the new phylogenetic subgroups identified among the Spanish B. bassiana s.s. isolates as Eu-7, Eu-8 and Eu-9. The results obtained from MP analyses (Figure 2), using a 1.1 kb fragment of the EF1-α gene from 56 isolates from our collection, confirmed that 53 isolates were B. bassiana s.s. (A), and three isolates grouped in three different phylogenetic subgroups within B. cf. bassiana (C). As in a previous study [7], the collection of Spanish isolates of B. bassiana s.s. was separated in five phylogenetic subgroups.

The three strains of sub-group 2 were isolated from Oceania (one from Australia and two from Papua New Guinea). To these, an Indian (Cfa), a Chinese (Cfa) and a Spanish (Csa) strain were also added, i.e., fungal strains from regions with temperate humid subtropic and Mediterranean

selleck inhibitor climates, resembling the climate of the Oceanic Cfa [41]. Sub-groups 3 and 4 consisted almost exclusively of European strains (9 and 3, respectively) from regions with Mediterranean climate, such as Spain, Portugal and Italy. On the other hand, 12 strains from regions of Europe with maritime temperate climates (Cfb) formed a well-supported group (87 and 92% NJ and MP bootstrap and 94% PP support) presented as sub-group 6. All nine strains of sub-group 5 were from regions with dry arid, semiarid (BSh, BSk and BWk) and temperate (Csa and Csb) climates in Asia and Europe, while the South American (6)

from tropic (Af, Am and Aw) and dry arid/semiarid (BSh) climates may be named as sub-group 7. Figure 6 Grouping of B. bassiana sensu lato strains (Clade Α) as well as Clade C and A 2 , according to their geographic distribution, climate conditions and molecular data (concatenated OSI-027 datasets from ITS1-5.8S-ITS2, nad 3- atp 9 and atp 6- rns ). The 3 symbol Köppen-Geiger climate classification is as shown in Fig. 5. Discussion Fungal mt genome size shows high divergence among the Pezizomycotina, ranging from 100.3 Kb for Podospora anserina (NC_001329) to 24.5 Kb for the entomopathogen Lecanicillium muscarium (AF487277). Beauveria mt genomes sizes were similar to those of other Sitaxentan fungi of the order Hypocreales, e.g., Fusarium oxysporum (34.5 Kb; AY945289) and Hypocrea jecorina (42.1 Kb; NC_003388), but they were significantly larger (~40%) than the mt genomes of the other two known entomopathogenic fungi of the order, i.e., M. anisopliae (24.7 kb) [27] and L. muscarium (24.5 kb) [42].

Since the Beauveria mtDNAs contained the same protein and rRNA coding genes -also identical in sizes- with all above mt genomes, their larger sizes can be attributed to more introns and to longer intergenic regions. Compared to mt genomes of plants and animals, fungal mt genomes are significantly richer in group I and II introns [43]. Divergence in intron content is a common feature among mt genomes of Pezizomycotina. At one extreme is the mt genome of P. anserina which contains 41 introns [44] and at the other are several fungi that contain a single intron in the rnl genes of their mt genomes (i.e., L. muscarium and M. anisopliae). The recently released mt genome of another B. bassiana isolate (EU371503) also presented fewer introns than the genomes that we analyzed. These data support and extend previous evidence for intronic variability among strains of the same Beauveria species [14, 16].

In addition, levels of activated caspase-3 and caspase-9 were significantly higher in cells treated with Photosan-II loaded in nanoparticles than free Photosan-II. Finally, treatment with nanoscale photosensitizers increased mouse survival and reduced tumor volume in mice to a greater extent compared with free photosensitizers. Overall, our data indicate that hollow nanoparticles IDO inhibitor containing photosensitizers more efficiently inhibit hepatoma cells than free photosensitizers, through induction of apoptosis, both in vivo and in vitro. Methods Cell lines The HepG2 human hepatoma cell line was purchased

from the cell center of the Xiangya School of Medicine of Central South University. Experimental animals Specific pathogen-free (SPF)-grade female BALB/c nude mice (26 to 30 days, 18 to 22 g) were obtained from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences. Mice were housed in SPF-grade animal laboratory of the Second Xiangya Hospital of Central South University in a temperature and humidity controlled room with food and water ad libitum. All procedures were approved

by the Animal Ethical Committee of Second Xiangya Hospital of Central South University. Preparation ACP-196 of nanoscale photosensitizers Nanoscale photosensitizers were prepared using a one-step wet chemical-based synthesis at room temperature, as previously described [15]. Tetraethyl orthosilicate (TEOS, 99.99%), polyacrylic acid (PAA, M.W = 3,000) were purchased from Aladdin Chemistry Co. Ltd (Shanghai, China). Anhydrous ethanol (99.7%) and ammonia (25% to 28%) were purchased from Sinopharm Chemical Reagent Co. Ltd (China) and Photosan-II (C34H38N4NaO5) obtained from Seehof Laboratorium F&E GmbH (Wesselburenerkoog, Germany). The resulted nanoscale photosensitizers (Photosan-II-loaded

also hollow silica nanospheres, 10 mg/L) showed good sphericity and narrow diameter distribution, ranging from 25 to 90 nm (mean value 37.8 nm). The encapsulation efficiency reached 95%. Cell culture and passaging Cryopreserved HepG2 human hepatoma cells were thawed and cultured in appropriate volume of 10% fetal bovine serum (FBS) in Dulbecco’s modified Eagle’s medium (DMEM) purchased from Gibco (USA), at 37°C and 5% CO2. Cell growth was observed daily, and culture media were changed as needed. Cells grown to logarithmic phase were trypsinized and passaged. MTT assay Two hundred microliters of a 105 cells/mL suspension was seeded into a 96-well plate and cultured as described above. Photosensitizers used were either conventional Photosan or nanoscale Photosan.