The FOXA1 DNA-binding domain structurally mimics the linker histone, H1, and stably binds to nucleosomal DNA, probably through interactions with the core histones, H3 and H4. These characteristics are associated with slow nuclear diffusion, abundant non-specific nucleosomal interactions, and stable binding at some Forkhead recognition motifs followed by nucleosome displacement see more and accessibility of surrounding regulatory DNA to other transcription

factors.[16, 17] Although the critical functions of Th cell master regulator transcription factors TBET and GATA3 have been well established for over a decade,[18-20] mechanistic insights and global, genomic characterization have been recent. How do Th cell master regulator transcription factors function and how extensive is their transcriptional and regulatory footprint? What are their roles in de novo enhancer activation and gene expression? Through what mechanisms do they modulate the activity of the regulatory elements that they bind – as bona fide pioneer factors displacing nucleosomes, through co-operative binding with other factors,

or through binding to previously accessible, poised elements? Early studies demonstrated the sufficiency of over-expressed TBET check details and GATA3 to induce DNase I accessibility and transcription at the interferon-γ (Ifng) and Th2 cytokine loci, respectively, and suggested their role in regulation of chromatin. In some cases this activity was shown to be independent of signals from cytokine receptors and downstream signal transducer and activator of transcription (STAT) factors or despite alternative lineage cytokine stimulation.[18, 19, 21-23] Loss of function studies established a requirement for these factors in Th differentiation in vivo.[20, 24] Importantly, these studies focused exclusively on small sets of signature Th1 and Th2 genes, usually the respective cytokine gene loci, and clearly established the important role of TBET

and GATA3 in their regulation. C-X-C chemokine receptor type 7 (CXCR-7) Subsequently, master regulators were described for Treg (FOXP3) and Th17 (RORγt) cells and shown to be critical for differentiation and acquisition of their respective T-cell lineage transcriptional programmes and phenotypes.[25-29] Their defining roles in CD4 T-cell subset differentiation and requirement for signature gene expression, analogous to classical master regulator transcription factor function, implied that Th master regulator transcription factors act as pioneer factors in the nucleation of de novo enhancer accessibility and activation. Recent studies suggest a model (Figs 1 and 2) that contrasts with this view, in which master regulators have limited footprints and act through collaboration with signal-activated environmental response factors.

Other techniques such as cyanoacrylates, fibrin glues, the Medtronic™ U-Clip®, and laser bonding have low levels of evidence supporting their use. Further research is required to establish any role for these techniques. © 2011 Wiley Periodicals, Inc. Microsurgery, 2012. “
“There is an increasing demand for successful free tissue transfer, with postoperative Selleck MK-2206 monitoring of flaps a key to early salvage. Monitoring methods have ranged from clinical techniques to invasive options, of which two are particularly applicable to buried flaps (Cook-Swartz Doppler probe and microdialysis). The evidence for these options has been represented largely in separate cohort studies,

with no single study comparing these three techniques. We aim to perform this comparison in a single cohort of patients. A prospective, consecutive cohort study comparing clinical monitoring, microdialysis and the implantable Doppler probe was undertaken. In 20 patients receiving 22 flaps, 21 flaps were monitored with microdialysis, 18 flaps with clinical observation, and 21 flaps with the Cook-Swartz Implantable Doppler probe. Exclusion was based on applicability and availability intra-operatively. Efficacy was assessed through PLX 4720 sensitivity, specificity, positive, and negative predictive values. Nineteen of 22 flaps had no suspected anastomotic problems; 3 of 22 flaps were explored for anastomotic

problems, with two salvaged and one lost. The implantable Doppler and microdialysis were found to detect flap statistically earlier than clinical assessment, with microdialysis better at detecting flap compromise: 100% specificity (confidence 4��8C interval 31–100%) when compared to the implantable probe and clinical assessment

(67%: 13–98% and 33%: 2–87%, respectively). Each of the Cook-Swartz Doppler probe, microdialysis and clinical assessment was found suitable for monitoring in free tissue transfer. The implantable Doppler and microdialysis offer the potential for earlier detection of flap compromise. © 2014 Wiley Periodicals, Inc. Microsurgery, 2014. “
“Local or distant metastatic recurrence after therapy is observed in 20–30% of cases of head-and-neck cancer. An unfavorable course may occur after cervical lymph node dissection due to loss of immunoprotective lymph nodes in the head-and-neck region. To overcome this problem, we performed autologous lymph node transplantation from the groin after head-and-neck cancer resection and cervical lymph node dissection. The patient was a 63-year-old man with squamous cell carcinoma in the mesopharyngeal lateral wall. After tumor resection and right cervical lymph node dissection, a lymph node-containing superficial circumflex iliac artery perforator flap was transplanted from the left groin. Pathological examination showed that cancer had invaded the primary tumor tissue stump. Thus, radiotherapy (66 Gy) was performed for the residual tumor from days 28 to 84 after surgery.

In contrast, none of the LPS-treated males developed diabetes (Fig. S1). Initiation of the treatment in NOD females at 12 weeks Dasatinib molecular weight of age, when mononuclear infiltration of Langerhans islets is readily detectable ([48] and not shown), also prevented progression to diabetes (Fig. 1B). However, administration of LPS after positive scoring for diabetes did not revert disease (data not shown). We next tested shorter LPS treatments. A single LPS injection into 7.5-week-old NOD females delayed diabetes onset by an average of 7 weeks but was not sufficient to significantly decrease diabetes incidence (Fig. 1C). Finally,

administration of LPS in 4-week-old female mice for 1 month resulted in 15 weeks delay in diabetes progression as compared with age-matched PBS-injected controls (Fig. 1D). We conclude that LPS is a potent inhibitor of diabetes occurrence in NOD mice.

The finding that continuous exposure to LPS protects 3-deazaneplanocin A in vitro NOD mice from diabetes, even after extensive infiltration of the pancreatic islets, suggests that LPS prevents insulitis progression. Our evidence that interruption of LPS treatment systematically leads to reactivation of disease, and hence diabetes establishment, supports the notion that the LPS effect is transient and it is exerted by maintaining diabetogenic T cells at check. Thereafter, to perform the cellular and functional analysis of LPS-protected NOD females, we chose the robust and long-lasting weekly regimen initiated in 6- to 8-week-old mice (Fig. 1A). It is still not known why few NOD females do not spontaneously progress to diabetes while they all reach Pyruvate dehydrogenase the stage of insulitis. Yet, it is well established that female NOD mice raised in germ-free conditions all develop disease [49]. Therefore, it was conceivable that LPS treatment would mimic an environmental factor of bacterial

origin present, although limited, in our SPF conditions. This reasoning prompted us to compare the two types of disease-free animals, namely LPS-treated and spontaneously protected, in what concerns sub-clinical signs of autoimmunity (Fig. 2A, B). To this aim it was necessary to focus our analysis on rather old animals (5–6 months of age), to increase the odds that the untreated normoglycemic controls were indeed spontaneously protected animals. In a first step, we evaluated whether the protective regimen affected directly the degree of islet infiltration. As expected, the majority of the islets in diabetic females presented severe infiltration; moreover, islet destruction was evident as indicated by a low number of detectable pancreatic islets (data not shown). Strikingly, LPS-treated mice were indistinguishable from age-matched healthy controls, as the majority of islets were devoid of infiltrates (60% and 66%, respectively), while the remaining islets displayed various degrees of infiltration, from mild to severe.

4A). We conclude that a strong passive saturating binding of IgE to basophils occurs in IgE knock-in mice in vivo. The central experiment to demonstrate a function of increased IgE in allergy is the analysis of anaphylaxis. Nutlin-3a datasheet The three genotypes (Fig. 3A) allowed a dissection of IgE versus IgG1 sensitizing capacity in an active anaphylaxis experiment. We used the same protocol for immunizing IgE knock-in mice as explained above (Fig. 3B), followed by an i.v. challenge with 30 μg TNP-OVA to induce systemic anaphylaxis (Fig. 4B). PBS-injected control mice did react with minimal body temperature drop of 0.5°C (Fig. 4B, panel2).

In sensitized IgEwt/ki and IgEki/ki mice, a comparable strong drop in body temperature of 6°C was observed, whereas WT mice reacted with moderate temperature drop of 4°C. It is important to note that the drop in body temperature in the IgE knock-in mice is more sustained compared with that in WT mice (Fig. 4B, panel 1). Surprisingly, only in the group of the

IgEki/ki mice, about 40%, died due to anaphylaxis (Fig. 4C panel 1). IgEki/ki p38 MAPK activity lack IgG1 and express high levels of antigen-specific IgE, yet are more susceptible to anaphylactic shock compared with WT mice, which express high levels of antigen-specific IgG1, but little IgE. Therefore, we suggest that antigen-specific IgE is a more potent inducer of anaphylaxis compared with antigen-specific IgG1. Mannose-binding protein-associated serine protease Importantly, while the IgEki/ki and the IgEwt/ki mice had similar temperature curves, death occurred only in the IgEki/ki mice, arguing for the strongest anaphylactic reaction in the IgEki/ki mice. These results argue against a major role for the alternative pathway of systemic anaphylaxis, which is mediated largely through IgG1 and FcγRIII and basophil activation in our model [8, 9]. In the following experiment, we addressed two questions, namely, whether CD23, the low affinity receptor for IgE, on B cells in conjunction with the IgE knock-in affects the

outcome of systemic anaphylaxis, and if basophil depletion influences IgE-mediated active anaphylaxis. First, we backcrossed the IgE knock-in mice to CD23-deficient mice [23]. No significant effect of a loss of CD23 on anaphylaxis in the IgEwt/wt animals was observed (Fig. 4B panel 2, open squares) when compared with the CD23 competent IgEwt/wt mice (Fig. 4B panel 1, open triangles). Also, no CD23-deficient mice died due to anaphylaxis (Fig. 4C panel 2), similar to wild-type animals (Fig. 4C panel 1). The double-mutant CD23−/− IgE knock-in heterozygous and homozygous mice respond to the anaphylactic challenge with faster and more sustained temperature drop and death (Fig. 4B and C, panels 3 and 4). Again, homozygous CD23−/− IgEki/ki mice display the strongest increase in lethality.

rubrum find more and T. mentagrophytes. Between 1995 and 2000 there were stated small differences in the number of isolated strains of dermatophytes in comparison with the number of examined patients. Since 2006 there has been observed a decrease in number of patients in our hospital with suspected fungal infections, but per cent of positive cultures has remained unchanged in comparison with earlier period. “
“Worldwide prevalence of non-dermatophyte mould onychomycosis has increased in recent years; however, available information on the topic is confusing and oftentimes contradictory, probably due to the small number of reported

cases. The aim of this study was to determine and describe the aetiological agents, as well as the epidemiological and clinical characteristics of non-dermatophyte mould

onychomycosis in a dermatology referral centre in Bogota, Colombia. A cross-sectional descriptive study was conducted between January 2001 and December 2011 among patients who attend the National Institute of Dermatology with a confirmed diagnosis of onychomycosis by non-dermatophytes moulds. There were 317 confirmed cases of non-dermatophyte mould onychomycosis in 196 women and 121 men whose average age was 43 years. Twenty-seven per cent of them had a history of systemic disease. The habit of walking and showering barefoot was PI3K inhibitors in clinical trials the major infection-related factor. Distal and lateral subungual presentation was the most common pattern of clinical presentation. The most frequent non-dermatophyte mould was Neoscytalidium dimidiatum followed by Fusarium spp. No relationship was observed with predisposing factors previously reported in the literature. Clinical features found in this population are indistinguishable from onychomycosis caused by dermatophytes. High

prevalence of N. dimidiatum found here was in contrast to a large number of studies where other Oxymatrine types of moulds predominate. “
“Simultaneous infections with multiple fungi may be misinterpreted as monomicrobial infections by current diagnostics with ramifications for the choice of antimicrobial agents that may impact patient outcomes. The application of molecular methods on tissue samples may be useful to decipher the aetiology of mixed fungal infections. We present a leukaemic patient who died from sepsis due to candidaemia. The postmortem examination documented fungal elements in lung tissue. Fungal DNA was amplified from the lung sample by broad-range PCR assays targeting the 28S ribosomal RNA gene or the internal transcribed spacer 2 (ITS-2). Fluorescence in situ hybridisation (FISH) using differentially labelled fungal probes was applied on the tissue. Sequencing identified the PCR amplicons as Aspergillus fumigatus (28S assay) and Candida tropicalis (ITS-2 assay).

Approval for human research was obtained from both the Human Vismodegib in vitro Investigation Committee at Wayne State University and the Ethics Committee

at the University of Cape Town (UCT) Faculty of Health Sciences. The 107 infants and mothers included in this study of effects on symbolic play are all those for whom complete data were available on the 17 prenatal alcohol exposure, play, and sociodemographic variables examined here. All women who reported drinking during pregnancy were advised to stop or reduce their intake, and all mothers were invited to participate in a home visitor intervention.1 The mother and child were transported by a staff driver and research nurse at 6.5, 12, and 13 months and 5 years selleck products to our laboratory at the UCT Faculty of Health Sciences, where the maternal interviews and neurobehavioral assessments were performed. At 5 years they were also transported to the FASD diagnostic clinic, which was held at a neighborhood church. Each mother was re-interviewed antenatally and at 1-month postpartum regarding her pregnancy alcohol and drug use. Interviews were conducted in Afrikaans or English,

depending on the mother’s preference. Each mother–infant dyad was provided breakfast prior to the assessments and interviews. All infant assessments were conducted and coded by research staff who were blind with respect to maternal alcohol use and group status; all maternal interviews including the Home Observation for Measurement of the Environment (HOME) were conducted by a developmental pediatrician (C. Progesterone D. Molteno) or research staff member who did not observe the infant cognitive or play assessments. In the initial timeline follow-back interview administered at recruitment in the MOU, the mother was asked about her drinking on a day-by-day basis during a typical 2-week period around the time of conception, with recall

linked to specific times of day activities. If her drinking had changed since conception, she was also asked about her drinking during the past 2 weeks and when her drinking had changed. At the follow-up antenatal visit in our laboratory, the mother was asked about her drinking during the previous 2 weeks. If there were any weeks since the recruitment visit when she drank greater quantities, she was asked to report her drinking for those weeks as well. At the 1-month postpartum visit, the mother was asked about her drinking during a typical 2-week period during the latter part of pregnancy, as well as her drinking during any weeks during that period when she drank greater quantities. Volume was recorded for each type of alcohol beverage consumed and converted to oz of AA by using the weights proposed by Bowman, Stein, and Newton (1975; liquor—0.4, beer—0.04, wine—0.2).

A mixture of two strains from

two different lactobacilli species (a combination of a high IL-10- and a GSK-3 beta phosphorylation high IL-12-inducing strain) was included in this study, but no clear synergistic effects were observed when compared with the individual strains. Although synergism is not always observed, some multispecies probiotic mixtures could expand the capacity for immunological modulation beyond that of the individual strains and might be effective in their immunomodulatory activity in selected patients (Timmerman et al., 2007; Niers et al., 2009; Kim et al., 2010; Lavasani et al., 2010). Summarizing the differential cytokine production profiles of the tested strains, it was observed that specific strains

selected on their IL-10-inducing properties, could be further grouped by their cytokine activity profile based on IFN-γ-inducing properties. The first group (represented by strains B1836, B2261, the mixture of B2261 and B633, B633 alone and CBI 118) induced a stronger proinflammatory TNF-α response, had a better inducing capacity of the Th1 compartment and showed a better inhibition of the Th2 cell compartment compared with the other group (represented by strains B1836 and B223), and might therefore be more appropriate candidate probiotics for allergic patients. A remarkable finding was the consistent inhibition of proliferation of hPBMC stimulated for 4 days with αCD3/αCD28 in the presence of all the applied bacterial strains with the most profound and significant inhibition by strain B633 in all seven allergic donors tested. ABT-199 concentration However, addition of strain Oxymatrine B633 did not impair cytokine induction, which strengthens the notion that various aspects of immunomodulation can be unique for a strain and that large species and strain differences exist in effects on inhibiting allergic inflammation, repression of hypersensitivity reactions and clinical symptoms of allergy (Medina et al., 2007; Isolauri & Salminen, 2008; Vissers et al.,

2010). These data support the notion that the probiotic potential of lactic acid bacteria as antiallergic compounds is strain specific and largely variable already in vitro as is also reported upon in vivo use in randomized double-blind, placebo-controlled clinical studies (Isolauri & Salminen, 2008; Kalliomaki et al., 2010). Donors with a documented pollen allergy were recruited outside the pollen season, resulting in a low frequency of allergen-specific T cells that can be as low as 1 per 20 000 cells (Gabrielsson et al., 1997), consequently resulting in a low response to the Bet v 1 allergen. Enrichment of the allergen-specific T cells or the use of allergen-specific T-cell clones would be necessary to study potential modulatory effects of bacterial strains under allergen-specific culture conditions (Ebner et al., 1995; Bohle, 2007).

The renal transport function was assessed by the uptake of para-aminohippurate SB431542 research buy (PAH) mediated organic anion transporters 1 (rOat1) and 3 (rOat3), using renal cortical slices. These two transporters were mainly expressed in renal proximal tubules and play important role for renal secretory process. Results: Comparing to T2DM, CGE supplemented

rats had significantly improved hyper-glycemia, hypertriglyceridemia, and insulin resistance. The uptake of PAH mediated by Oat1 and 3 functions in renal slices was not different among experimental groups. Interestingly, CGE blunted sodium nitroprusside-induced impairment of PAH uptake in T2DM rats. This data also correlated with the levels of NO accumulation in renal cortical tissues. Conclusion: These findings indicated that CGE has anti-diabetic effect and prevents diabetic nephropathy partly through nitrosative stress

selleck inhibitor pathway. HASEGAWA KAZUHIRO, WAKINO SHU, HAYASHI KOICHI, ITOH HIROSHI Department of Nephrology, Keio University, Tokyo, Japan Introduction: Sirtuin 1 (Sirt1), a NAD-dependent deacetylase with positive effects on cellular and whole-body metabolism, is expressed in the renal cortex and medulla. Among various renal cells, we previously reported that proximal tubular Sirt1 plays pivotal roles (Hasegawa K, BBRC 2008, JBC 2010). Sirt1 is also known to have protective effects against diabetic damages in liver or pancreas. However, a correlation between renal Sirt1 and diabetic kidney damages has not been investigated. Therefore, we aim to investigate the role of Sirt1 in diabetic nephropathy (DN). Methods and Results: We found that Sirt1 in proximal tubules (PTs) was downregulated before albuminuria, and, thereafter, Sirt1 in podocytes (Pods) was downregulated in DN mice including both streptozotocin-induced and obese (db/db) mice. Then, we created PT-specific Sirt1 transgenic (Tg) and conditional knockout (CKO) mice to examine the role of PT’s Sirt1. Sirt1 Tg prevented and CKO aggravated VAV2 glomerular changes

and albuminuria that occured in diabetes, respectively. Non-diabetic CKO mice exhibited albuminuria, suggesting that Sirt1 in PTs affects glomerular function. We also observed that reduced PT’s Sirt1 in DN decreased NMN (Nicotinamide Mono Nucleotide, a key intermediate of Sirt1-related nicotinic acid metabolism) led to decreasing Pod’s Sirt1. Reduced Sirt1 increased Claudin-1, a tight junction protein, in Pods by an epigenetic mechanism whereby decreased Pod’s Sirt1 inactivated Dnmt1 leading to reduced CpG methylation of Claudin-1 gene, which contributed to increased Claudin-1 expression and albuminuria. Intriguingly, Claudins are generally known to strengthen the epithelial barrier, but we novely showed that overexpression of Claudin-1 in Pods increased glomerular permeability by activating β-catenin–Snail pathway.

However, we found that SIRPα was rapidly induced on Kupffer cells following L. donovani infection, via a mechanism

involving G-protein-coupled receptors. Thus, we describe a novel amplification pathway affecting cytokine production by hepatic iNKT cells, which may facilitate the breakdown of hepatic tolerance after infection. “
“The transfer of alloreactive regulatory T (aTreg) cells into transplant recipients represents an attractive treatment option to improve long-term graft acceptance. We recently described a protocol for the generation of aTreg cells in mice using a nondepleting anti-CD4 antibody (aCD4). Here, we investigated whether adding TGF-β and retinoic acid (RA) or rapamycin (Rapa) can further improve aTreg-cell generation and function. Murine CD4+ T cells were cultured with allogeneic B cells in the

presence of aCD4 alone, aCD4+TGF-β+RA or aCD4+Rapa. GPCR Compound Library cell assay Addition of TGF-β+RA or Rapa resulted in an increase of CD25+Foxp3+-expressing T cells. Expression of CD40L and production of IFN-γ and IL-17 was abolished in aCD4+TGF-β+RA aTreg cells. Additionally, aCD4+TGF-β+RA aTreg cells showed the highest level of Helios and Neuropilin-1 co-expression. Although CD25+Foxp3+ cells from AG-014699 in vivo all culture conditions displayed complete demethylation of the Treg-specific demethylated region, aCD4+TGF-β+RA Treg cells showed the most stable Foxp3 expression upon restimulation. Consequently, aCD4+TGF-β+RA aTreg Methane monooxygenase cells suppressed effector T-cell differentiation more effectively in comparison to aTreg cells

harvested from all other cultures, and furthermore inhibited acute graft versus host disease and especially skin transplant rejection. Thus, addition of TGF-β+RA seems to be superior over Rapa in stabilising the phenotype and functional capacity of aTreg cells. Regulatory T (Treg) cells play an important role in the suppression of unwanted immune responses after transplantation [1] or after allogeneic stem cell transplantation [2]. Treg cells are essential for maintaining peripheral tolerance and for preventing autoimmune diseases such as systemic lupus erythematosus [3], rheumatoid arthritis [4] or diabetes [5]. Treg cells can be categorised into two groups, natural Treg (nTreg) cells, which develop in the thymus [6], and adaptive Treg cells or so-called induced (iTreg) Treg cells, which develop from CD4+CD25− cells in the periphery. Treg cells are mainly characterised by their expression of CD4 and CD25 [7]. Although both subsets express the fork head transcription factor Foxp3, nTreg cells and iTreg cells differ in DNA methylation pattern of the Foxp3 gene [8]. Furthermore, nTreg cells have been shown to express the Ikaros transcription family member Helios [9], although the selectivity of Helios expression in thymus-derived Treg cells was recently challenged [10].

Lu et al. have suggested that recipient-derived MCs are crucial for Treg-mediated peripheral tolerance [11], indicating that the function of mast cells in suppressing immune responses was related to Tregs. Our study showed that CD4+CD25+ FoxP3+ cells could be induced by BMMCs. This finding may supply a new mechanism suggesting that MCs are crucial for Treg-mediated transplant tolerance [11]. This method may also become a new method for the induction of Tregsin vitro. Our results showed that the highest percentage of Tregs was found in the highest ratio (2:1) of BMMCs to T cells. TGF-β1 expression in BMMCs was determined in our experimental

groups. Jahanyar et al. concluded that mast cell-derived TGF-β may serve as important mediators for Treg activation in allografts [21], and other studies reported that the percentage of Tregs increased with the higher level of added TGF-β1 [22]. Therefore, it seems that the increase of Tregs with a higher ratio of BMMCs HM781-36B price may be related to more BMMCs-derived TGF-β1. Consistent with previous studies, and in order to test whether BMMC-derived TGF-β1 is involved in Carfilzomib chemical structure the generation of Tregs, TGF-β1

neutralizing antibody was added to the co-culture system [4]. The conversion of Tregs was reduced significantly by the TGF-β1 neutralizing antibody, but the TGF-β1 neutralizing antibody could not reverse Treg induction completely. The percentages of Tregs were still higher than control, even with the application of TGF-β1 neutralizing antibody. Whether there were some other mediators derived from BMMCs which also had the potential to induce Tregs is debatable. Metz considered that IL-4 may be related to the suppression function of MC in the immune response [6]. Therefore, IL-4 neutralizing antibody was applied to block the function of IL-4, but there were no significant differences after the application of IL-4 neutralizing antibody.

Although this study did not provide direct evidence for BMMCs as the main source of TGF-β1, it suggests that BMMC-derived TGF-β1 is involved in the regulation of Treg cell generation in vitro. Our experiment concerned mainly the relationship between mast cells and Tregsin vitro. Huang et al. showed that tumour-infiltrating mast cells may promote tumour growth through one way of increasing Treg cells in vivo[23]. Demeclocycline This leads us to conclude that perhaps Tregs can be induced by mast cells in vivo. More studies will be conducted to clarify this phenomenon. In conclusion, our experiments demonstrate that Tregs can be induced by BMMCs in vitro, and secreting TGF-β1 by BMMCs is one of the principal factors for the effect. This finding may provide new evidence that mast cells have the ability to suppress immune responses by way of Treg induction. Furthermore, the study may supply new data for identifying clearly the role of mast cells in immune systems. This work was funded by National Natural Science Foundation of China (no.