The cultured cells were randomly divided into three groups: control group (0 Gy, without the embedded seed in the paraffin), 2 Gy, and 4 Gy. Apoptosis analysis by flow cytometry Adherent SW 1990 cells cells were trypsinized

and centrifuged for 5 min at 220xg. Cells were then washed three times in ice cold PBS and suspended in binding selleck inhibitor buffer (0.01 M Hepes, pH 7.4; 0.14 M NaCl; 2.5 mM CaCl2) at 1 × 106 cells/ml. The cells were stained with annexin V-FITC (1 μl/ml) and propidium iodide (5 μg/ml) for 15 min in the dark as described previously [15]. Cells were analyzed by fluorescence-activated cell sorting (FACS) using a Coulter EPICS and MOdFit SOFTWARE (Verity Software House, Topsham, MN). Each test was performed 3 times. Real-time polymerase chain reaction (PCR) Total RNA was retracted from SW 1990 cells using Trizol

reagent (Invitrogen, Carlsbad, CA). The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase GAPDH was used as an internal reference [16]. Real-time PCR was performed by using the following primers: find more for DNMT1, upstream primer 5′-GTGGGGGACTGTGTCTCTGT-3′ downstream primer 5′-TGAAAGCTGCATGTCCTCAC-3′, and amplified fragment length of 204 bp; for DNMT3a, upstream primer 5′-ATCTCGCGATTTCTCGACTC- 3′, downstream primer 5′-GCTGAACTTGGCTATCCTGC -3′, and amplified fragment length of 180 bp; for DNMT3b, upstream primer 5′-TTGAATATGAAGCCCCCAAG- 3′, downstream primer selleck kinase inhibitor 5′-TGATATTCCCCTCGTGCTTC -3′, amplified fragment length of 160 bp; for GAPDH, upstream primer 5′-GCACCGTCAAGGCTGAGAAC-3′, downstream primer 5′-ATGGTGGTGAAGACGCCAGT-3′, amplified fragment length of 142 bp. Cycling parameters: pre-denaturation 1 min, 95°C; denaturation 15 s, 95°C;

annealing 15 s, 60°C; extension 45 s, 72°C, 40 cycles; final extension 5 min, 70°C. The PCR was repeated three times for each sample. The standard curve was generated with the ABI 7500 Real Time PCR system (Applied Biosystems, Carlsbad, CA, USA) to describe the linear relationship between threshold cycle (Ct) value and relative quantity (RQ). RQ values were obtained from measured Ct value with the following formula: 2(-ΔΔCt), where ΔΔCt = ΔCtT; ΔCtS = (ΔCtT – ΔCtTE ) – (ΔCtS – ΔCtSE), T is the target sample, S is the SW-1990 cell sample, and E is the reference. The RQ of mRNA in all groups were calculated relative to the RQ value in control group 1. Western blotting Western blotting Inositol monophosphatase 1 was performed as described previously [17, 18]. Nuclear protein was prepared from SW-1990 pancreatic cancer cells with a Nuclear Protein extraction kit (Fermentas, Ontario, CA). The total protein concentration was determined by the Bradford assay using the Coomassie Protein Assay Reagent Kit (Pierce Biotechnology, Rockford, IL). Prepared protein samples (20 μg each) were boiled for 5 min and loaded onto a 12% SDS polyacrylamide gel. After separation by electrophoresis and electroblotting to nitrocellulose membranes, membranes were blocked by with 5% nonfat dry milk in 0.

CA Cancer PI3K inhibitor J Clin 2005, 55: 242–258.Adriamycin CrossRefPubMed 23. O’Sullivan B, Shah J: New TNM staging criteria for head and neck tumors. Semin Surg Oncol 2003, 21: 30–42.CrossRefPubMed 24. Liao WT, Wang X, Xu LH, Kong QL, Yu CP, Li MZ, Shi L, Zeng MS, Song LB: Centromere protein H is a novel prognostic marker for human nonsmall cell lung cancer progression and overall patient survival. Cancer 2009, 115: 1507–1017.CrossRefPubMed

25. Jane C, Nerurkar AV, Shirsat NV, Deshpande RB, Amrapurkar AD, Karjodkar FR: Increased survivin expression in high-grade oral squamous cell carcinoma: a study in Indian tobacco chewers. J Oral Pathol Med 2006, 35: 595–601.CrossRefPubMed 26. Kops GJ, Weaver BA, Cleveland DW: On the road to cancer: aneuploidy and the mitotic checkpoint. Nat Rev Cancer 2005, 5: 773–785.CrossRefPubMed 27. de la Guardia C, Casiano CA, Trinidad-Pinedo J, Baez A: CENP-F gene amplification and overexpression in head and neck squamous cell carcinomas. Head Neck Trichostatin A nmr 2001, 23: 104–112.CrossRefPubMed 28. Clark GM, Allred DC, Hilsenbeck SG, Chamness GC, Osborne CK, Jones D, Lee WH: Mitosin (a new proliferation marker) correlates with clinical outcome in node-negative breast cancer. Cancer Res 1997, 57: 5505–5508.PubMed 29. Carvalho A,

Carmena M, Sambade C, Earnshaw WC, Wheatley SP: Survivin is required for stable checkpoint activation in taxol-treated HeLa cells. J Cell Sci 2003, 116: 2987–2998.CrossRefPubMed 30. Lens SM, Vader G, Medema RH: The case for Survivin as mitotic regulator. Curr Opin Cell Biol 2006, 18: 616–622.CrossRefPubMed 31. Altieri DC: Survivin, versatile modulation of cell division and apoptosis in cancer. Oncogene 2003, 22: 8581–8589.CrossRefPubMed 32. Li F, Ambrosini G, Chu EY, Plescia J, Tognin S, Marchisio PC, Altieri DC: Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 1998, 396: 580–584.CrossRefPubMed 33. Lo Muzio L, Farina A, Rubini C, Pezzetti F, Stabellini G, Laino G, Santarelli A, Pannone G, Bufo P, de Lillo A, Carinci F: Survivin as prognostic factor in

squamous cell carcinoma of the oral cavity. Cancer Lett 2005, 225: 27–33.CrossRefPubMed 34. Lo Muzio L, Pannone G, Leonardi R, Staibano S, Mignogna MD, De Rosa G, Kudo Y, Takata T, Altieri DC: Survivin, a potential early predictor of tumor progression in the oral mucosa. J Dent Res 2003, 82: 923–928.CrossRefPubMed Pembrolizumab in vivo Competing interests The authors declare that they have no competing interests. Authors’ contributions WL carried out cell cultures, establishment of stable cell lines, proliferation functional assays, and preparation of manuscript. CY and DW participated in RT-PCR and immunohistochemistry, as well as data analysis. LX and GW have been involved in western blot analysis and data interpretation. LZ participated in critical revision of the manuscript. MZ participated in the study design and coordination and helped to revise the manuscript.

In addition, polyamines (spermine and spermidine) inhibit the production of tumoricidal cytokines, such as tumor necrosis factor (TNF), and chemokines in vitro, while they do not inhibit production of transforming check details growth factor beta, which has immunosuppressive properties [105–107]. Conversely, in animal experiments, polyamine deprivation has been shown to enhance chemokine production, reverse tumor inoculation-induced inhibition of killer cell activity, and prevent tumor-induced immune suppression [108, 109]. TNF is able to induce apoptotic cell death and to attack and destroy www.selleckchem.com/products/sgc-cbp30.html cancer cells

[110], while LFA-1 and CD56, especially bright CD11a and bright CD56 cells, are required for the induction of LAK cell cytotoxic activity [111, 112]. Polyamines suppress LAK cytotoxicity without decreasing cell viability and activity in vitro, and the changes in blood spermine levels are negatively associated with changes in LAK cytotoxicity in cancer patients

[42]. 6. Sources of polyamines other than cancer cells learn more Food is an important source of polyamines. Polyamines in the intestinal lumen are absorbed quickly and distributed to all organs and tissues [29, 39, 40]. Moreover, continuous intake of polyamine-rich food gradually increases blood polyamine levels [30, 31]. Therefore, the restricted intake of food polyamine and inhibition of polyamine synthesis by microbiota in the intestine with or without inhibitor-induced inhibition of polyamine synthesis is reported to have favorable effects on cancer therapy [33, 113–115]. Trauma, such as surgery, is itself considered to increase the risk

of cancer spread through various mechanisms [116–118]. Blood concentration and urinary excretion of polyamines are known to increase after surgery, although the origin of this increase is not well established [97, 119]. Our previous study showed that increases in blood polyamine levels are inversely associated with anti-tumor LAK cytotoxicities in patients who have undergone surgery [42]. In addition to mechanisms previously postulated for post-traumatic cancer spread, buy Y-27632 post-operative increases in polyamines may be another factor that accelerates tumor growth. Conclusion As polyamines are essential for cell growth, one of the mechanisms by which polyamines accelerate tumor growth is through the increased availability of this indispensable growth factor. In addition, polyamines seem to accelerate tumor invasion and metastasis not only by suppressing immune system activity against established (already existing) tumors but also by enhancing the ability of invasive and metastatic capability of cancer cells.

C 1 ′ and C 2 ′ are background currents. To fit the photocurrent curves when the linearly polarized direction of the incident light is along [1 0], [110], [100] and [010] selleck compound crystallographic directions, selleck chemicals llc respectively, we find that parameters S 1, S 1 ′ and S 1 − are considerably larger than parameters S 2, S 2 ′, S 2 ±, S 3, S 3 ′ and S 3 ±. The detailed fitting

results of the parameters are listed in Table 1. This reveals that polarization independent currents are dominant in total magneto-photocurrents. Furthermore, we found that the parameters S 1 and S 1 ′ are slightly smaller than S 1 −. The polarization-independent currents present anisotropy of crystallographic directions. The parameters of linearly polarized light-induced photocurrents are in the same order of magnitude except the S 3 is larger. Table 1 Fitting selleck kinase inhibitor results of the parameters   Value S 1 5.535 S 2 −0.015 S 3 0.383 S 1 ′ −5.241 S 2 ′ −0.003 S 3 ′ 0.018 S 1 + 0.269 S 1 − −6.093 S 2 + −0.016 S 2 − −0.015 S 3 +

0.002 S 3 − −0.018 Units: . From the microscopic point of view, the electric photocurrent density can be calculated by summing the velocities of the photo-excited carriers. The magneto-photocurrent in μ direction (μ=x,y) can be described by [5, 22] (5) e is the electron charge. denotes the electron velocity along μ direction. In the excitation process, is the steady-state nonequilibrium photo-excited electron density in Zeeman-splitting conduction bands. It can be described by Equation 6 for the linearly polarized radiation. (6) ϕ is the angle between the wave vector and the x direction. α is the angle between the plane of linear polarization and the x direction. Considering the contribution of asymmetric relaxation of electrons to the current, we should

add an additional term to the . Then the in Equation 6 includes contributions Cell press of both excitation and relaxation. Owing to the magneto-photocurrent in this superlattice is independent of the radiation polarization, it can be deduced that is much larger than and . This conclusion is similar to that in [22] which that reported always overwhelms and theoretically. The radiation polarization independent of MPE generated by direct interband transition had also been observed in the BiTeI film [23]. However, in (110)-grown GaAs/Al x Ga 1−x As quantum wells, MPE generated by indirect intrasubband transition shows clear relations to the radiation linear polarization state [24]. The reason may be that in the intrasubband transition process, spin-dependent asymmetric electron-phonon interaction which contributes to the magneto-photocurrent is sensitive to the radiation polarization state. It leads to the relative magnitudes of and in Equation 6 increase. More practically, the phonon effect may be taken into account when designing optically manipulated spintronics devices in the future.

Secondly, in the naïve diversity profile for the putative methyltransferase group, the lines representing the diversity of the 2007A, 2009B, and 2010B samples crossed each

other numerous times between q = 0 and q = 5 (Additional file 1: Figure S2). Lastly, in the naïve profile for the putative concanavalin A-like glucanases/lectins group, the 2010B samples were as diverse as or more diverse than the 2007A samples at q = 0, but the diversity of 2010B samples dropped sharply and remained lower than all other samples after approximately q = 0.5 (Additional file 1: Figure S3). In the case of viral diversity, ultra-rare taxa play an important role in rapid evolution to allow new viruses to infect hosts that are constantly evolving defense Tanespimycin mechanisms. Thus, diversity calculated at low values of q, which are sensitive to rare taxa, is the more appropriate measure of viral diversity. Figure 1 Hypersaline lake viruses Cluster 667 diversity profiles. (A) Naïve and (B) similarity-based (phylogenetic relatedness) diversity profiles calculated for Cluster 667 from the hypersaline lake viruses data. We see similar STI571 nmr results

for the acid mine drainage dataset. At q = 0 (species richness) in the naïve analysis, the Env-3 at growth stage 2 check details sample is the most diverse sample, but the sample’s diversity decreases and is surpassed by the growth stage 0 bioreactor sample and both Env-1 samples between Morin Hydrate q = 1 and q = 2 (Figure 2), demonstrating that the bioreactor and Env-1 samples were less even than the Env-3 sample at growth stage 2. Thus, for this dataset as well as for the hypersaline lake viruses dataset, evaluating the diversity of the microbial communities at multiple values of q leads to a different interpretation of the results and response to the original hypotheses (Table 1). Figure 2 Acid mine drainage bacteria and archaea (HiSeq) diversity profiles. (A) Naïve and (B) similarity-based (phylogenetic relatedness) diversity profiles calculated from the acid mine drainage bacteria and archaea HiSeq data. Diversity profiles do not always add new information

to analyses of natural microbial datasets. In some cases, such as with the naïve profiles of the subsurface bacteria dataset, the most diverse samples in a dataset were always calculated as the most diverse, across the entire range of q in the naïve profile (Figure 3). Thus, whether we quantified diversity using species richness, Shannon diversity, or diversity profiles, we would arrive at the same result. In general, our findings provide evidence for the utility of diversity profiles to analyze microbial datasets, even when similarity information is not taken into account, because they allow researchers to visualize multiple diversity indices across the range of q in the same place after just one calculation.

Thus, the cHtrA N-terminal

signal peptide is sufficient for directing PhoA across the bacterial inner membrane. We further found that the secretion of cHtrA was not inhibited by the C1 compound, an inhibitor known to inhibit chlamydial type III secretion system [52]. As positive controls, C1 inhibited the secretion of both IncA and CT621, two known chlamydial type III secretion substrates [30, 52]. Consistently, the secretion of CPAF was not affected by C1. This is because secretion of CPAF is dependent on type II secretion pathway this website [62]. Figure 7 cHtrA is secreted via a sec-dependent pathway. (A) The SignalP 3.0 program with both the Neural Networks (NN) and Hidden Markov Model (HMM) algorithms http://​www.​expasy.​ch was used to analyze the precursor cHtrA sequence from C. trachomatis serovar D http://​stdgen.​northwestern.​edu/​. The NN algorithm predicts a signal peptide from the first methionine residue (M1) to a serine residue at position 16 (S16) while the HMM-predicted signal peptide is M1-S23. (B) The M1-S23 peptide of cHtrA (cHtrAss) Dinaciclib cost directed translocation of PhoA into bacterial periplasmic

space (cHtrAss-’PhoA, slot 1, blue). Expression of the positive control full-length PhoA construct also led to the translocation of mature PhoA (with its intrinsic signal peptide, slot 3, blue) but the negative control mature PhoA construct failed to do so (without a signal peptide, ‘PhoA, slot selleck chemicals llc 2, white). (C) Bacterial transformants expressing the same three constructs were fractionated into periplasmic Sorafenib supplier (per) and cytosolic (cyto) fractions and the fractions were detected with antibodies against a FLAG tag (anti-Flag, panel a) and GroEL (anti-GroEL, panel b) on a Western blot. Mature PhoA was secreted into the periplasm of bacteria expressing either the full-length PhoA construct

or HtrAss-PhoA construct while mature PhoA stayed in the cytoplasm of the bacteria expressing the mature PhoA alone construct. (D) cHtrA secretion into the cytosol of chlamydia-infected cells is not inhibited by the type III secretion inhibitor C1 compound. HeLa monolayers infected with C. trachomatis L2 for 6 hr were treated with DMSO (panels a, c & e) or 50 μM C1 (b, d & f). Thirty-six hours after treatment, the cultures were processed for triply labeling with antibodies against IncA (green) and cHtrA, CT621 or CPAF (red) and DAPI for DNA (blue). C1 inhibited secretion of IncA and CT621 but not cHtrA or CPAF. Red arrows indicate chlamydia proteins that are secreted into host cell cytosol. Discussion The obligate intracellular growth of Chlamydia requires the organisms to intimately interact with host cells.

1% Triton X-100 for 15 min and blocked in 3% H2O2-methyl alcohol for 15 min. The coverslips were Epacadostat ic50 incubated with anti-IDH1 rabbit polyclonal antibody (protein technology group, USA) in blocking buffer overnight at 4°C. Coverslips were then incubated with an anti-rabbit secondary antibody and peroxidase-conjugated strepavidin-biotin complex (Santa Cruz, CA, USA) at 37°C for 45 min at room temperature in the dark [23]. Immunoreactivity was visualized with diaminobenzidine (DAB) (Zymed, South San Francisco, CA). Negative controls were obtained by omitting the primary antibody. Slides were scanned

using a microscopy (Carl Zeiss AG, Germany), images were recorded using a digital camera (DC 500, Leica) and the Leica FW 4000 software and images were processed using Adobe Photoshop.

Real-time PCR Cellular total RNA from OS cells was extracted with TRIZOL Reagent (Invitrogen, Carlsbad, CA, USA). The concentration of RNA was determined by the absorbance at 260 nm and the purity was determined by the 260/280 ratio with a BioPhotometer(Eppendorf, Hamburg, Germany). For each reaction, 1 μg RNA was reverse-transcribed selleck inhibitor with random primer by ReverTra Ace (Toyobo, Osaka, Japan). RNA quality and efficiency of reverse transcription were examined by PCRs from each 1 μl cDNA according to the manufacturer’s recommendations [24]. The mRNA expression of IDH-1, p53 and internal control geneβ-actin was quantified by Real-time PCR Detection System (SLAN, HONGSHI) with SYBR Green I (Toyobo, Osaka, Japan). As PCR was performed according to standard procedures [24, 25] after optimization, PCR-reactions were within the exponential range of amplification. MycoClean Mycoplasma Removal Kit The gene-specific exon-spanning PCR primer pair for IDH1 was 5′-TCAGTGGCGGTTCTGTGGTA-3′,5′-CTTGGTGACTTGGTCGTTGGT-3′, and for p537-8 was 5′-CAGCCAAGTCTGTGACTTGCACGTA C-3′,5′-CTATGTCGAAAAGTGTTTCTGTCATC-3′, and for β-actin was 5′-GTCCACCGCAAATGCTTCTA-3′,5′-TGCTGTCACCTTCACCGTTC-3′. The sequences of the primers were checked by Nucleotide BLAST for specific gene amplification. Omission of cDNA template was used as a negative control. Triplicate measurements

were made of all genes in each patient and data of mean were used. For VRT752271 clinical trial relative quantification of genes expression level, standard curves were built by considering at least three points of a ten-fold dilution series of cDNA in water. Relative gene expression data are given as the n-fold change in transcription of the target genes normalized to the endogenous control in the same sample. Protein extraction and Western blot Lysates of cells were prepared using lysis buffer from the Dual-Luciferase assay kit (Promega) according to the manufacturer’s recommendations. The lysates were collected and centrifuged at 12,000 g for 10 min at 4°C. The protein in the supernatants were pooled together and stored at -80°C until concentration analyzed by the BCA Protein Assay Kit (Sangon, Shanghai, China).

Finally, the cells, wells, and membranes were washed with PBS. For FACS analysis, the cells were fixed with 2% p-formaldehyde. Then absorbance at 450 nm (ELISA), chemiluminescence (dot-blotting analysis), or fluorescence (FACS; Excalibur, Beckton Dickinson) were detected. Biofilm formation Homotypic biofilm formation by P. gingivalis was performed as described by others [50]. Briefly, P. gingivalis cells were grown on ABA plates, then in BM supplemented with hemin or dipyridyl to OD660 = 1.0 and used to inoculate fresh cultures to OD660 = 0.1. Cells in the appropriate medium were transferred (200

μl) into sterile round-bottom microtiter plates (Sarstedt) and incubated under anaerobic conditions at 37°C for 24 or 48 h. The resulting biofilms were washed with PBS, stained with Proteasome activity 1% selleck crystal violet, washed with PBS, and de-stained with 96% ethanol. Absorbance (A) was determined at 570 nm using a Multiskan Ascent microplate reader. The assays were repeated at least three times with each strain Adavosertib price grown in eight wells. To confirm that the P. gingivalis cells were viable, the biofilm cells were scrapped into the respective medium and the OD at 660 nm and colony-forming

unit (CFU) values were evaluated after 24 and 48 h (see Additional file 3). In parallel, bacteria were grown in planktonic form and the OD at 660 nm and CFU values were measured after 24 and 48 h. Growth and biofilm inhibition studies Bacteria were grown overnight on ABA plates and then in BM supplemented with hemin or dipyridyl to OD660 = 1.0. After centrifugation, the bacteria were washed and suspended in PBS to OD660 = 0.1. Then

5 ml of the bacterial suspension was centrifuged and the bacteria were incubated in 200 μl of PBS for 1 h at 37°C with the IgG fraction purified from pre-immune or immune anti-HmuY rabbit serum (200 ng). After addition of 5 ml of the appropriate medium, planktonic bacterial growth was monitored by measuring the OD at 660 nm or biofilm formed as described above. Assays were performed three times in duplicate. new Statistical analysis Data are expressed as means values ± standard deviations (mean ± SD). Statistical analysis was performed using unpaired Student’s t test (GraphPad Prism 5). Values of p < 0.05 were considered statistically significant. Acknowledgements This work was supported in part by grant nos. N401 029 32/0742, N N303 406136, and N N303 518438 from the Ministry of Science and Higher Education, and by Wroclaw Research Center EIT+ under the project “”Biotechnologies and advanced medical technologies – BioMed”" (POIG 01.01.02-02-003/08/00) financed from the European Regional Development Fund (Operational Program Innovative Economy, 1.1.2) (TO) and the European Social Fund (Human Capital Program, 8.2.

anthracis.

Determined by the Selleckchem JNK inhibitor analysis of 14 canSNP sites described by Van Ert et al[5]. The five canSNP groups represented in China are indicated in larger and bold fonts in this Neighbor Joining Tree. The number of isolates (N), genotypes (G), and Nei’s Diversity Index [8] within groups (D) are illustrated in the table in the lower left. Neighbor-joining trees based upon additional MLVA genotypes within each of these 5 canSNP groups are illustrated in Figures 3 and 5. The basic tree is now defined by 7 sequenced genomes that form 7 sub-branches or sub-lineages ending in “”stars”" in Figure 1. Each of these sub-lineages is designated by the nomenclature from the whole genome sequence site in Genbank, e.g. A.Br Ames, A.Br.WNA (for western North America), and A.Br.Vollum. The relative position of each canSNP is indicated by vertical OSI-906 in vitro script and a small

arrow and is arbitrarily HDAC inhibitor defined, e.g., as A.Br.001 where A refers to the major subgroup and 001 is the first canSNP (see the A.Br.Ames sub-lineage in Figure 1, also [5]). In this case the derived A.Br.001 SNP defines all isolates that are on the same branch as the sequenced Ames strain. In addition to these 7 sub-lineages the analysis of 26 diverse isolates uncovered 5 nodes or sub-groups along the branches of this tree. Four of these nodes are in the major A Branch and one is in the B Branch (see “”circles”" in Figure 1). These nodes are defined by the two canSNPs on either side of the node position, e.g. A.Br.001/002 or A.Br.008/009. All of the initial 1,000 isolates in the Van Ert study [5] were placed into one and only one of these 12 sub-lineages or sub-groups. Results CanSNP analysis of isolates from China The 191 B. anthracis isolates from China were distributed into only five of these 12 canSNP sub-lineages/sub-groups described

by Van Ert et al. [5]. These canSNP groups were A.Br.Vollum, A.Br.Aust.94, A.Br.001/002, A.Br.Ames, and A.Br.008/009 (Figures 1 and 2). Four of the sub-lineages/sub-groups (A.Br.Vollum, A.Br.Aust.94, A.Br.008/009 and A.Br.001/002) were found in the western province of China, Xinjiang (Figure 2). But only isolates from A.Br.001/002 see more sub-group and the close relative A.Br.Ames sub-lineage were found scattered throughout the other regions of China from east to west. These findings clearly suggest 4 or 5 separate introductions of B. anthracis into or out of China, with 3 possibly involving the routes defined as the Silk Road. Figure 2 Geographical distribution of B. anthracis isolates in China. This distribution is based on 12 canSNP genotypes described in Figure 1 and the analysis of 191 isolates from China; also see [5]. The red routes include the western City of Kashi in Xinjiang Province, the main crossroads into China and around the Taklimakan Desert leading into the eastern Chinese provinces.