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Immune checkpoint blockade (ICB) therapies have achieved significant breakthroughs in cancer treatment over the past decade. However, ICB is largely ineffective in desert-type gastric cancer (GC) due to intrinsic tumor heterogeneity and a highly immunosuppressive tumor microenvironment (TME). Transforming tumors from immunosuppressive to immunostimulatory is a potential approach to enhance ICB response. Here, we developed a chromosomal instability (CIN) subtype GC mouse model with an immunoactive TME and a stem cell-originated mouse-derived allograft (MDA) model with an immunosuppressed TME to investigate mechanisms regulating the tumor immunophenotype and uncover therapeutic strategies to remodel the TME. Blocking β-catenin signaling attenuated the immunochemotherapeutic resistance of MDA tumors. The tyrosine kinase inhibitor apatinib reprogrammed the TME by increasing CD8+ T cells and IGHA+ plasma cells infiltration and decreasing M2 macrophages, but apatinib also induced PD-L1 and CD80 expression in both human and mouse desert-type tumors. Oxaliplatin decreased the apatinib-induced expression of immune checkpoints and enhanced the antitumor efficacy of immunotherapy. A prospective clinical trial (NCT04195828) demonstrated that a neoadjuvant regimen of apatinib plus ICB and chemotherapy was effective in patients with desert-type GC. Collectively, these findings identify potential drug targets for immune desert-type GC driven by β-catenin signaling.

p-Phenylenediamines (PPDs) are widely used as antioxidants in numerous rubber products to prevent or delay oxidation and corrosion. However, their derived quinones (PPD-Qs), generated through reactions with ozone, are ubiquitous in the environment and raise significant health and toxicity concerns. This review summarizes the current state of knowledge on environmental distribution and fate, human exposure, and biological toxicity of PPDs and PPD-Qs, and makes recommendations for future research directions. Although PPDs and PPD-Qs have been monitored in a variety of environmental matrices, studies on soil, sediment, and organisms remain limited. This shortcoming hinders our understanding of their distribution patterns and migration mechanisms in these specific environments. These contaminants can enter the human body through various exposure routes, but toxicological studies have not yielded sufficient results to derive risk thresholds for the assessment of human health. Most studies examining biological and toxicological effects have focused on acute exposure scenarios, which do not accurately reflect the long-term interactions that occur in natural settings. The toxic effects of PPDs and PPD-Qs on zebrafish, nematodes, and mammals include neurobehavioral changes, reproductive dysfunction, and digestive damage, which are linked to mitochondrial stress, DNA adduct formation, and disrupted lipid metabolism, respectively. However, the underlying toxicological mechanisms remain poorly understood. Future research should prioritize the investigation of the impacts of PPDs and PPD-Qs on various organizational levels within biota to provide a scientific basis for developing effective risk management measures., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Objective: Acute cerebral infarction is a common complication of intracranial tuberculosis (TB), causing irreversible damage to brain tissue and significantly affecting patient prognosis. This study aims to explore the risk factors associated with acute cerebral infarction in patients with intracranial tuberculosis., Methods: We retrospectively analyzed data from eligible intracranial TB patients treated at our hospital between January 2020 and March 2023. Based on MRI findings, patients were categorized into a cerebral infarction group and a non-infarction group. Clinical data, cerebrospinal fluid (CSF) examinations, and imaging features (such as hydrocephalus, cerebral arteritis, and meningeal thickening) were compared between the two groups. Binary logistic regression analysis was used to identify risk factors for acute cerebral infarction in patients with intracranial TB., Results: A total of 102 patients were included, with 24 in the cerebral infarction group and 78 in the non-infarction group. Male patients accounted for 87.5% in the infarction group and 58.3% in the non-infarction group. Patients with a Glasgow Coma Scale (GCS) score of 3-10 accounted for 45.8% in the infarction group compared to 15.4% in the non-infarction group. The incidence of hydrocephalus, cerebral arteritis, and meningeal thickening was significantly higher in the infarction group (37.5, 54.2, and 79.2%, respectively) compared to the non-infarction group (6.4, 6.4, and 43.6%, respectively) ( p  < 0.05). The parenchymal type of intracranial TB was less frequent in the infarction group (20.8%) than in the non-infarction group (56.4%), while the mixed type was more frequent in the infarction group (62.5%) compared to the non-infarction group (26.9%) ( p  < 0.05). Patients with meningeal thickening involving the cisterns and basal cisterns had a higher risk of cerebral infarction ( p  < 0.05). Multivariate binary logistic regression analysis revealed that male sex (OR = 13.56; 95% CI 1.25-38.30) and cerebral arteritis (OR = 19.32; 95% CI 0.94-37.64) were independent risk factors for cerebral infarction in intracranial TB patients., Conclusion: Male sex and the presence of cerebral arteritis are independent risk factors for acute cerebral infarction in patients with intracranial tuberculosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Huang, Qiu, Wang, He and Wang.)

Background: Nasopharyngeal carcinoma (NPC) is a malignant tumor associated with Epstein-Barr virus (EBV) infection. Chemoradiotherapy is the mainstream treatment for locally advanced NPC, and chemotherapeutic drugs are an indispensable part of NPC treatment. However, the toxic side-effects of chemotherapy drugs limit their therapeutic value, and new chemotherapy drugs are urgently needed for NPC. Silvestrol, an emerging natural plant anticancer molecule, has shown promising antitumor activity in breast cancer, melanoma, liver cancer, and other tumor types by promoting apoptosis in cancer cells to a greater extent than in normal cells. However, the effects of silvestrol on NPC and its possible molecular mechanisms have yet to be fully explored., Methods: Cell counting kit-8 (CCK-8), cell scratch, flow cytometry, 5-ethynyl-2'-deoxyuridine (EdU), and Western blot (WB) assays were used to evaluate the effects of silvestrol on the cell viability, cell cycle, apoptosis, and migration of NPC cells. RNA sequencing (RNA-Seq) was used to study the effect of extracellular signal-regulated kinase (ERK) inhibitors on the cell transcriptome, and immunohistochemistry (IHC) to assess protein expression levels in patient specimens., Results: Silvestrol inhibited cell migration and DNA replication of NPC cells, while promoting the expression of cleaved caspase-3, apoptosis, and cell cycle arrest. Furthermore, silvestrol altered the level of ERK phosphorylation. The ERK-targeted inhibitor LY3214996 attenuated silvestrol-mediated inhibition of NPC cell proliferation but not migration. Analysis of RNA-Seq data and WB were used to identify and validate the downstream regulatory targets of silvestrol. Expression of GADD45A, RAP1A, and hexokinase-II (HK2) proteins was inhibited by silvestrol and LY3214996. Finally, IHC revealed that GADD45A, RAP1A, and HK2 protein expression was more abundant in cancer tissues than in non-tumor tissues., Conclusions: Silvestrol inhibits the proliferation of NPC cells by targeting ERK phosphorylation. However, the inhibition of NPC cell migration by silvestrol was independent of the Raf-MEK-ERK pathway. RAP1A, HK2, and GADD45A may be potential targets for the action of silvestrol., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Chemical transfer across the air-water interface is one of the most important geochemical processes of global significance. Quantifying such a process has remained extremely challenging due to the lack of suitable technologies to measure chemical diffusion across the air-water microlayer. Herein, we present a fluorescence optical system capable of visualizing the formation of the air-water microlayer with a spatial resolution of 10 μm and quantifying air-water diffusion fluxes using pyrene as a target chemical. We show for the first time that the air-water microlayer is composed of the surface microlayer in water (∼290 ± 40 μm) and a diffusion layer in air (∼350 ± 40 μm) with 1 μg L -1 of pyrene. The diffusion flux of pyrene across the air-water interface is derived from its high-resolution concentration profile without any pre-emptive assumption, which is 2 orders of magnitude lower than those from the conventional method. This system can be expanded to visualize diffusion dynamics of other fluorescent chemicals across the air-water interface and provides a powerful tool for furthering our understanding of air-water mass transfer of organic chemicals related to their global cycling.

Background: The heterogeneous crosstalk between tumor cells and other cells in their microenvironment means a notable difference in clinical outcomes of head and neck squamous cell carcinoma (HNSCC). CD8+ T cells and macrophages are effector factors of the immune system, which have direct killing and phagocytosis effects on tumor cells. How the evolution of their role in the tumor microenvironment influences patients clinically remains a mystery. This study aims to investigate the complex communication networks in the HNSCC tumor immune microenvironment, elucidate the interactions between immune cells and tumors, and establish prognostic risk model., Methods: 20 HNSCC samples single-cell rna sequencing (scRNA-seq) data and bulk rna-seq data were derived from public databases. The "cellchat" R package was used to identify cell-to-cell communication networks and prognostic related genes, and then cell-cell communication (ccc) molecular subtypes were constructed by unsupervised clustering. Kaplan-Meier(K-M) survival analysis, clinical characteristics analysis, immune microenvironment analysis, immune cell infiltration analysis and CD8+T cell differentiation correlation analysis were performed. Finally, the ccc gene signature including APP, ALCAM, IL6, IL10 and CD6 was constructed based on univariate Cox analysis and multivariate Cox regression. Kaplan-Meier analysis and time-dependent receiver operating characteristic (ROC) analysis were used to evaluate the model in the train group and the validation group, respectively., Results: With CD8+T cells from naive to exhaustion state, significantly decreased expression of protective factor (CD6 gene) is associated with poorer prognosis in patients with HNSCC. The role of macrophages in the tumor microenvironment has been identified as tumor-associated macrophage (TAM), which can promote tumor proliferation and help tumor cells provide more nutrients and channels to facilitate tumor cell invasion and metastasis. In addition, based on the strength of all ccc in the tumor microenvironment, we identified five prognostic ccc gene signatures (cccgs), which were identified as independent prognostic factors by univariate and multivariate analysis. The predictive power of cccgs was well demonstrated in different clinical groups in train and test cohorts., Conclusion: Our study highlights the propensity for crosstalk between tumors and other cells and developed a novel signature on the basis of a strong association gene for cell communication that has a powerful ability to predict prognosis and immunotherapy response in patients with HNSCC. This may provide some guidance for developing diagnostic biomarkers for risk stratification and therapeutic targets for new therapeutic strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Wang, Sun, Cao, Hu, Qian, Jiang, Jiang, Zhou, Qiu and Wang.)

Background: We report on a case of Vibrio vulnificus (V. vulnificus) detected by metagenomics next-generation sequencing (mNGS) in a 53-year-old male patient with polymicrobial gas gangrene and successful treatment by surgery. This report raises awareness among dermatologists that when a patient is clinically suspected of a special type of pathogenic infection, the mNGS method should be preferred to identify the patient's pathogen infection as soon as possible and then take effective treatment in time to save patients' lives., Case Summary: A 53-year-old male who worked in the aquatic market complained of redness and swelling of the lower limbs, blisters and ulcers with fever for 3 d. We used mNGS to test the pathogens in ulcer secretions. The results were returned in 24 h and indicated: V. vulnificus , Fusobacterium necrophorum , Staphylococcus haemolyticus , Staphylococcus aureus , Streptococcus dysgalactiae and Klebsiella aerogenes . This patient was diagnosed with V. vulnificus infection. The emergency operation was performed immediately under combined lumbar and epidural anesthesia: Left leg expansion and exploration (August 10, 2021). After surgery, we continued to use piperacillin sodium tazobactam sodium 4.5 g every 8 h and levofloxacin 0.5 g for anti-infection treatment. The patient underwent further surgery under lumbar anesthesia on August 17, 2021 and August 31, 2021: Left leg deactivation and skin grafting, negative pressure closed drainage and right thigh skin removal. After treatment, the transplanted flap survived., Conclusion: We could confirm the diagnosis of Vibrio vulnificus infection within 24 h through mNGS detection and then immediately performed emergency surgery., Competing Interests: Conflict-of-interest statement: All the authors report having no relevant conflicts of interest for this article., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Despite ample evidence on spreading of e-waste derived hazardous materials, riverine transport of organic contaminants from e-waste recycling zones to surrounding areas has not been evaluated. To address this issue, passive and grab sampling methods were used to assess sediment-water diffusion and horizontal transport of polybrominated diphenyl ethers (PBDEs) and organophosphorus flame retardant (OPFRs) at upstream and downstream sites of two rivers in a typical e-waste recycling zone. Sediment acted as a source of BDE-17 with fluxes of 0.007-0.04 ng m -2 d -1 at all sampling sites. BDE-47 and BDE-99 reached equilibrium between overlying water and sediment porewater. Sediment interconverted from a sink at the upstream site to a source of OPFRs at the downstream site with a flux varying between -7.3 and 234 ng m -2 d -1 . The amounts of OPFRs (11-45 g d -1 ) via horizontal riverine transport were greater than those of PBDEs (0.68-2 g d -1 ). The vertical sediment-water diffusion of PBDEs and OPFRs was not significant compared to horizontal riverine transport. The annual riverine outputs of PBDEs and OPFRs from the downstream sites were 250-330 g and 12,000-16,500 g, respectively, indicating the significance of riverine transport of organic contaminants from e-waste recycling zones to surrounding areas., (Copyright © 2022. Published by Elsevier Ltd.)

Cutaneous squamous cell carcinoma (CSCC) is one of the most common types of skin cancer in humans worldwide. The identification and characterization of cancer-associated transmembrane proteins are important for understanding the molecular biology of CSCC. The aim of the present study was to evaluate the expression pattern of transmembrane protein 40 (TMEM40) in CSCC and its clinical significance. The underlying mechanisms were also examined. Reverse transcription-quantitative PCR, western blot and immunohistochemistry analysis were used to determine the relative expression of TMEM40 in CSCC cell lines and clinical tissue samples. The effect of TMEM40 gene silencing on cell proliferation was also evaluated using Cell Counting Kit-8 assays. Wound healing assays, flow cytometry and Transwell assays were used to explore the migration, cell cycle distribution/apoptosis and invasion of CSCC cells following TMEM40 silencing, respectively. In the present study, increased TMEM40 expression was observed in CSCC tissue samples, compared with normal skin, and TMEM40 expression was associated with large tumor size in patients with CSCC. In vitro functional assays indicated that TMEM40 was involved in the regulation of A431 and SCL1 cell growth through its effects on the cell cycle and apoptosis. Silencing TMEM40 in A431 and SCL1 cells resulted in cell cycle arrest at the G 0 /G 1 phase and promoted apoptosis. In addition, migration and invasion were significantly inhibited following silencing of TMEM40 expression in CSCC cells. Taken together, the results of the present study indicated that reduced TMEM40 expression could inhibit CSCC development and that TMEM40 may represent a therapeutic target in CSCC., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Yu et al.)

The partition coefficient (K pew ) of an analyte between low-density polyethylene (LDPE) film and water is a critical parameter for measuring freely dissolved concentrations of the analyte with PE passive sampling devices. Measuring log K pew for super hydrophobic organic chemicals (HOCs) have been proven extremely difficult. The present study developed a large volume model for measuring log K pew of super HOCs, i.e., novel halogenated flame retardants (NHFRs). Results showed that the measured log K pew values of selected PAHs and PCBs obtained by the large volume model were in line with those from the co-solvent model and the literature data within less 0.3 log units of difference, while those of NHFRs (6.27-7.34) except for hexachlorocyclopentadienyldibromocyclooctane (HCDBCO) and Decabromodiphenyl ethane (DBDPE) were significantly lower than those (6.51-8.89) from the co-solvent model. A curvilinear relationship was observed between log K pew and log K ow of all target compounds, with the turning point at log K ow  = ∼8.0 in the large volume model, but that was not found for the co-solvent model. These can be attributed to the large molecular volumes (> 450 Å 3 ) for NHFRs, which require high Gibbs free energy to penetrate into the inside structures of LDPE in the large volume model. However, the solvent swelling effects in the co-solvent model needs to be investigated. Therefore, the large volume model is robust to determine the K pew values of super HOCs for facilitating the application of aquatic passive sampling techniques., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

1. Overexpression of P-glycoprotein (P-gp, encoded by MDR1) mediates resistance to multiple immunosuppressors. Several common MDR1 variants (1236C > T, 2677G > T, 3435C > T) impact the efflux activity of P-gp-mediated substrates. We assessed the effect of these polymorphisms on the sensitivity, intracellular accumulation, and efflux of tacrolimus, cyclosporine A, sirolimus and everolimus in transfected LLC-PK1 cells. 2. LLC-PK1 cell lines were transfected with empty vector (pcDNA3.1) and recombinant MDR1 T-T-T , MDR1 C-T-T , MDR1 C-G-T and MDR1 C-G-C vectors, respectively and further screened in the presence of puromycin. The IC 50 values, intracellular accumulation, and apparent permeability ratios of tacrolimus, cyclosporine A, sirolimus and everolimus were evaluated. 3. MDR1 overexpression increased the resistance of LLC-PK1 cells to tacrolimus, cyclosporine A, sirolimus and everolimus. The resistance of cells expressing MDR1 C-G-C wild-type haplotypes to tacrolimus were increased compared to MDR1 T-T-T , MDR1 C-T-T , MDR1 C-G-T variant haplotypes. The efflux ability of P-gp-mediated tacrolimus in cells transfected with MDR1 C-G-C was higher than cells overexpressing MDR1 T-T-T , MDR1 C-T-T , MDR1 C-G-T variant haplotypes. In addition, the resistance of cells expressing MDR1 C-G-C wild-type haplotypes to sirolimus were increased compared to MDR1 C-T-T , MDR1 C-G-T variant haplotypes. The efflux ability of P-gp-mediated sirolimus in cells overexpressing MDR1 C-G-C was higher than cells transfected with MDR1 C-T-T , MDR1 C-G-T variant haplotypes. 4. These findings indicate that wild-type MDR1 exports tacrolimus and sirolimus more efficiency than the MDR1 T-T-T , MDR1 C-T-T , MDR1 C-G-T variant protein. This observation indicates that 1236C > T, 2677G > T, 3435C > T variant haplotypes drastically decrease the efflux ability of P-gp-mediated tacrolimus and sirolimus in a substrate-specific manner.

Purpose: Free flap compromise after internal jugular venous thrombosis (IJVT) occurs fairly infrequently in oral and maxillofacial rehabilitation and reconstruction but could seriously affect the patient's postoperative recovery. The aim of this study was to analyze and discuss the characteristics and management of such flap compromise., Patients and Methods: The authors performed a retrospective case series of 13 patients who underwent reconstruction of oral and maxillofacial defects with anterolateral thigh flaps and developed flap compromise from IJVT from July 2014 through December 2016 at the Second Xiangya Hospital of Central South University (Changsha, China). The clinical features of flap compromise are described, and the precautions and improvement measures are reported., Results: All 13 patients were men with an average age of 50.3 years. Flap compromise caused by IJVT occurred on postoperative days 3 to 7, of which 2 flaps were salvaged (15.4%) and 11 flaps failed completely after immediate urgent re-exploration. Four patients presented with orocutaneous fistula after salvage surgery that healed well after a period of wound dressing., Conclusions: Because of the late occurrence and low salvage rate of flap compromise caused by IJVT, intensive flap monitoring after reconstruction surgery is necessary for patients with free flap transfer., (Copyright © 2018 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

The alterations in P-glycoprotein (P-gp)-mediated transport of antipsychotics due to the ABCB1 1199G>A polymorphism were assessed in the current study. The ABCB1 wt and ABCB1 1199A recombinant cell models were constructed to study the sensitivity, intracellular accumulation, and transepithelial permeability of antipsychotic drugs. ABCB1 1199A recombinant cells had more sensitivity to olanzapine (2.2-fold, p < 0.01), aripiprazole (1.8-fold, p < 0.01), amisulpride (2.3-fold, p < 0.01), and risperidone (3.1-fold, p < 0.01) than ABCB1 wt cells, while the resistance to paliperidone in both recombinant cell models was similar. In addition, the uptake quality of olanzapine, aripiprazole, amisulpride, and risperidone in ABCB1 1199A recombinant cells was greatly decreased compared to ABCB1 wt cells (3.2-fold, p < 0.01; 3.7-fold, p < 0.01; 3.1-fold, p < 0.01; 2.6-fold, and p < 0.01, respectively). Furthermore, apparent permeability values were greatly increased in ABCB1 1199A recombinant cells compared with ABCB1 wt recombinant cells for olanzapine (2.7-fold, p < 0.01), aripiprazole (2.9-fold, p < 0.01), amisulpride (3.4-fold, p < 0.01), and risperidone (4.1-fold, p < 0.01). The influence of ABCB1 1199G>A polymorphism on the transport of P-gp-mediated substrates showed up as drug-specific. Collectively, the ABCB1 1199G>A polymorphism may impact effective antipsychotics concentration in target cells via mediating the agents transport and distribution.

Oscillatory activity of retinal ganglion cell (RGC) has been observed in various species. It was reported such oscillatory activity is raised within large neural network and involved in retinal information coding. In the present research, we found an oscillation-like activity in ON-OFF RGC of bullfrog retina, and studied the mechanisms underlying the ON and OFF activities respectively. Pharmacological experiments revealed that the oscillation-like activity patterns in both ON and OFF pathways were abolished by GABA receptor antagonists, indicating GABAergic inhibition is essential for generating them. At the meantime, such activities in the ON and OFF pathways showed different responses to several other applied drugs. The oscillation-like pattern in the OFF pathway was abolished by glycine receptor antagonist or gap junction blocker, whereas that in the ON pathway was not affected. Furthermore, the blockade of the ON pathway by metabotropic glutamate receptor agonist led to suppression of the oscillation-like pattern in the OFF pathway. These results suggest that the ON pathway has modulatory effect on the oscillation-like activity in the OFF pathway. Therefore, the mechanisms underlying the oscillation-like activities in the ON and OFF pathways are different: the oscillation-like activity in the ON pathway is likely caused by GABAergic amacrine cell network, while that in the OFF pathway needs the contributions of GABAergic and glycinergic amacrine cell network, as well as gap junction connections., Competing Interests: The authors declare that they have no conflict of interest. Ethical standard All experimental procedures were performed according to the humane treatment and use of animals as prescribed by the Association for Research in Vision and Ophthalmology, and were approved by the Ethic Committee, School of Biomedical Engineering, Shanghai Jiao Tong University.

Pine wilt disease (PWD) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, is responsible for devastating epidemics in pine trees in Asia and Europe. Recent studies showed that bacteria carried by the PWN might be involved in PWD. However, the molecular mechanism of the interaction between bacteria and the PWN remained unclear. Now that the whole genome of B. xylophilus (Bursaphelenchus xylophilus) is published, transcriptome analysis is a unique method to study the role played by bacteria in PWN. In this study, the transcriptome of aseptic B. xylophilus, B. xylophilus treated with endobacterium (Stenotrophomonas maltophilia NSPmBx03) and fungus B. xylophilus were sequenced. We found that 61 genes were up-regulated and 830 were down-regulated in B. xylophilus after treatment with the endobacterium; 178 genes were up-regulated and 1122 were down-regulated in fungus B. xylophilus compared with aseptic B. xylophilus. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were used to study the significantly changed biological functions and pathways for these differentially expressed genes. Many pathogenesis-related genes, including glutathinone S-transferase, pectate lyase, ATP-binding cassette transporter and cytochrome P450, were up-regulated after B. xylophilus were treated with the endobacterium. In addition, we found that bacteria enhanced the virulence of PWN. These findings indicate that endobacteria might play an important role in the development and virulence of PWN and will improve our understanding of the regulatory mechanisms involved in the interaction between bacteria and the PWN.

As the causal agent of pine wilt disease (PWD), the pine wood nematode (PWN), Bursaphelenchus xylophilus, causes huge economic losses by devastating pine forests worldwide. The pectate lyase gene is essential for successful invasion of their host plants by plant-parasitic nematodes. To demonstrate the role of pectate lyase gene in the PWD process, RNA interference (RNAi) is used to analyze the function of the pectate lyase 1 gene in B. xylophilus (Bxpel1). The efficiency of RNAi was detected by real-time PCR. The result demonstrated that the quantity of B. xylophilus propagated with control solution treatment was 62 times greater than that soaking in double-stranded RNA (dsRNA) after B. xylophilus inoculation in Botrytis cinerea for the first generation (F1). The number of B. xylophilus soaking in control solution was doubled compared to that soaking in Bxpel1 dsRNA four days after inoculation in Pinus thunbergii. The quantity of B. xylophilus was reduced significantly (p < 0.001) after treatment with dsRNAi compared with that using a control solution treatment. Bxpel1 dsRNAi reduced the migration speed and reproduction of B. xylophilus in pine trees. The pathogenicity to P. thunbergii seedling of B. xylophilus was weaker after soaking in dsRNA solution compared with that after soaking in the control solution. Our results suggest that Bxpel1 gene is a significant pathogenic factor in the PWD process and this basic information may facilitate a better understanding of the molecular mechanism of PWD.

Background. We aimed to investigate the distributive characteristics of SLC22A1 rs594709 and SLC47A1 rs2289669 polymorphisms and their influence on metformin efficacy in Chinese T2DM patients. Methods. The distributions of SLC22A1 rs594709 and SLC47A1 rs2289669 polymorphisms were determined in 267 T2DM patients and 182 healthy subjects. Subsequently, 53 newly diagnosed patients who received metformin monotherapy were recruited to evaluate metformin efficacy. Results. No significant difference was found between T2DM patients and healthy subjects in SLC22A1 rs594709 and SLC47A1 rs2289669 allele frequencies and genotype frequencies. After metformin treatment, SLC22A1 rs594709 GG genotype patients showed a higher increase in FINS (p = 0.015) and decrease in HOMA-IS (p = 0.001) and QUICKI (p = 0.002) than A allele carriers. SLC47A1 rs2289669 GG genotype patients had a higher decrease in TChol (p = 0.030) and LDL-C (p = 0.049) than A allele carriers. Among SLC22A1 rs594709 AA genotype, patients with SLC47A1 rs2289669 AA genotype showed a higher decrease in FBG (p = 0.015), PINS (p = 0.041), and HOMA-IR (p = 0.014) than G allele carriers. However, among SLC22A1 rs594709 G allele carriers, SLC47A1 rs2289669 AA genotype patients showed a higher decrease in TChol (p = 0.013) than G allele carriers. Conclusion. Our data suggest that SLC22A1 rs594709 and SLC47A1 rs2289669 polymorphisms may influence metformin efficacy together in Chinese T2DM patients.

Congenital hemidysplasia with ichthyosiform nevus and limb defects (CHILD) syndrome is an X-linked autosomal dominant disorder characterized by unilateral congenital hemidysplasia with ichthyosiform erythroderma and ipsilateral limb defects caused by a mutation in the gene encoding NAD[P]H steroid dehydrogenase-like protein (NSDHL) at Xq28. The histopathologic hallmark of skin lesions in CHILD syndrome is psoriasiform epidermis with hyperkeratosis and parakeratosis, and its most striking feature affecting the upper dermis is filling of the papillary dermis with foam cells. Here we present the case of a 9-year-old Chinese girl born with the typical clinical features of CHILD syndrome. Histologic and immunohistochemical evaluation of the skin lesions confirmed the diagnosis and led to identification of a heterozygous point mutation in exon 8 of the NSDHL gene. In addition, we provide a literature review of 26 unrelated CHILD syndrome patients from different countries, caused by 20 unique gene mutations occurring throughout the entire NSDHL gene, to promote understanding and provide a more comprehensive description of this unusual disorder., (© 2015 Wiley Periodicals, Inc.)

Objective: To study the effect of Gan-Pi regulatory needling (GPRN) in treating chloasma and its influences on female sex hormones, superoxide dismutase (SOD), lipid peroxide (LPO) and melanocyte-stimulating hormone (MSH)., Methods: Ninety chloasma patients were equally randomized to three groups, the treatment group treated with GPRN, the control group treated with conventional Western medicine and the blank group untreated. Changes in the scores of skin lesion (area and color) and symptom, as well as blood levels of female sex hormones, MSH, SOD and LPO were observed and compared after 3 months of treatment., Results: In the treatment group, the scores of skin lesion area and color were reduced from 2.76 + or - 0.96 and 2.48 + or - 0.78 before treatment to 1.42 + or - 0.42 and 1.03 + or - 0.41 after treatment, respectively, while in the control group they were from 2.78 + or - 1.06 and 2.53 + or - 0.88 to 1.58 + or - 1.23 and 1.28 + or - 0.96, respectively, all showing significant changes (P<0.05); the scores were insignificantly changed in the blank group (P>0.05). At the same time, the score of symptoms in the treatment group significantly improved after treatment (P<0.05), significantly different from that of the other two groups. Comparison of female sex hormones among groups showed no significant differences either before or after treatment. The level of LPO decreased and SOD increased in both the treatment group and the control group significantly (all P<0.05), but significant lowering of MSH was only seen in the treatment group (P<0.05)., Conclusions: GPRN can effectively lessen the size and lighten the color of chloasma, improve the accompanying symptoms in patients and decrease LPO and MSH levels and increase the SOD level, but will not affect the level of the female sex hormones.

Here we show a simplified and improved method to produce large quantities of evenly distributed monolayer cultures that display major characteristics of adipocytes. These cultures are applicable for quantitative analysis for biochemical and molecular events in adipogenesis during development and may provide a useful system for high-throughput drug screening assays of antiobesity drugs. In our method, we treated embryoid bodies (EBs) with all-trans retinoic acid (ATRA) for 3 days, 1 day after they attached to the gelatin-coated culture plates without further transfer. The cells were maintained in insulin and trioiodothyronine (T(3))-containing medium until day 12, when they were dispersed by enzymatic digestion and replated onto multiple culture plates. Two days later, adipocyte induction factors were added for 6 days and examined 6 days later. The amount of lipid droplet-laden adipocytes in the culture reached approximately 80%, with a nearly five-fold increase in GPDH activity. The cells expressed high levels of adipose-specific proteins (adipocyte markers), including PPARgamma2, ALBP, LPL, HSL, perilipin, and DGAT1. The adipocytes are functionally active, as evidenced by their response to lipolytic agents, such as forskolin, Bt2-cAMP, and isoproterenol, with more than 20-fold increases in glycerol release.

Loss of genomic imprinting of insulin-like growth factor II (IGF2) is a hallmark of many human neoplasms. We attempted to correct this aberrant epigenotype by transferring nuclei from human tumor cells that showed loss of IGF2 imprinting into enucleated mouse and human fibroblasts that had maintained normal IGF2 imprinting. After nuclear transfer, the abnormal biallelic expression of IGF2 in tumor nuclei transiently converted to normal monoallelic imprinted expression in the reconstructed diploid cells. In tetraploid hybrid cells, however, normal IGF2 imprinting was permanently restored in the tumor genome. Inhibition of the synthesis of putative trans imprinting factors with cycloheximide led to loss of IGF2 imprinting in normal cultured fibroblasts, suggesting that normal cells produce proteins that act in trans to induce or maintain genomic imprinting. These data demonstrate that an abnormal tumor epigenotype can be corrected by in vitro reprogramming, and suggest that loss of imprinting is associated with the loss of activity of non-CTCF trans imprinting factor(s) that are either inactivated or mutated in tumors.

Gene transcription may be regulated by remote enhancer or insulator regions through chromosome looping. Using a modification of chromosome conformation capture (3C) and fluorescence in situ hybridization, we found that one allele of the insulin-like growth factor 2 (Igf2)/H19 imprinting control region (ICR) on chromosome 7 colocalized with one allele of Wsb1/Nf1 on chromosome 11. Omission of CCCTC-binding factor (CTCF) or deletion of the maternal ICR abrogated this association and altered Wsb1/Nf1 gene expression. These findings demonstrate that CTCF mediates an interchromosomal association, perhaps by directing distant DNA segments to a common transcription factory, and the data provide a model for long-range allele-specific associations between gene regions on different chromosomes that suggest a framework for DNA recombination and RNA trans-splicing.