The Association of Brain Insulin Resistance with Anesthesia/Surgery-Induced Cognitive Deterioration Is Female-Specific in 5XFAD Transgenic Mice.

Background: Our previous studies indicated that anesthesia/surgery could aggravate cognitive impairment and tau pathology in female 5XFAD transgenic (Tg) mice. However, it is unknown whether there are sex differences in the susceptibility of developing postoperative cognitive dysfunction in 5XFAD Tg mice. Objective: In this study, we aim to determine whether anesthesia/surgery can have different effects on female and male 5XFAD Tg mice, and to explore the underpinning mechanisms. Methods: The mice received abdominal surgery under isoflurane anesthesia. Morris water maze was used to assess the cognitive function. Hippocampal levels of p-tau (AT8), p-IRS1 (Ser612), IRS1, p-GSK3ß (Tyr216), and p-GSK3ß (Ser9) at postoperative day 1 were evaluated by western blot assays. Results: Anesthesia/surgery exaggerated cognitive impairment and tau pathology in female, but not male 5XFAD Tg mice. The anesthesia/surgery led to elevated hippocampus protein levels of p-IRS1 (Ser612)/IRS1 ratio and p-GSK3ß (Tyr216) and reduced hippocampus protein levels of p-GSK3ß (Ser9) in female, but not male 5XFAD Tg mice. Conclusions: This study demonstrated that female 5XFAD Tg mice were more susceptible to anesthesia/surgery-induced cognitive deterioration and tau pathology aggravation, potentially due to female-specific brain insulin resistance.

[Role of COX-2/PGE2/EP4 Axis-induced Macrophage Functional Activation 
in NSCLC Development].

BACKGROUND: Tumor microenvironment (TME) is one of the important factors in tumorigenesis and progression, in which tumor-associated macrophages (TAMs) play an important role in non-small cell lung cancer (NSCLC) progression. However, the mechanism of TAMs in NSCLC progression remains unclear, so this study aimed to investigate the role of TAMs in NSCLC progression and to find potential therapeutic targets. METHODS: Gene Expression Profiling Interactive Analysis (GEPIA) database was used to analyze the expression of prostaglandin E2 receptor 4 (EP4) mRNA in NSCLC and normal lung tissues; the protein expression levels of cyclooxygenase-2 (COX-2), EP4, cluster of differentiation 86 (CD86), CD163 and CD31 were detected by immunohistochemistry (IHC) in 120 NSCLC tissues and 24 paracancerous tissues specimens. The nude mouse lung adenocarcinoma cell A549 and macrophage RAW264.7 co-transplanted tumor model was established. And the samples were collected by gavage with EP4 inhibitor E7046, and then stained with hematoxylin-eosin (HE), IHC, and immunofluorescence (IF), and then detected by Western blot for the epithelial mesenchymal transformation (EMT) of the tumor tissues of the nude mice in each group. Western blot was used to detect the expressions of EMT related protiens in each group of nude mice; full-length transcriptome sequencing was used to screen the key genes causing liver metastasis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed. RESULTS: EP4 mRNA expression level in NSCLC tissues was generally lower than that in normal lung tissues (P<0.05); COX-2, EP4, CD163, CD31 proteins were differentially expressed in NSCLC tissues and adjacent tissues, and differences were observed in many clinicopathological parameters of NSCLC patients; RAW264.7 shortened the latency period of tumorigenesis of A549 and promoted the proliferation of tumors and liver metastasis of tumors, and E7046 could reduce tumor cell proliferation activity, tumor tissue vascular density and M2-type macrophage infiltration in nude mice; IF staining showed that macrophages were mainly distributed around the metastatic foci of tumors; Western blot results showed that compared with A549 alone transplantation group, the relative expression of E-cadherin protein in tumor tissues of mice in A549 and RAW264.7 co-transplantation group was significantly decreased, and the difference was statistically significant (P<0.05), while the relative expression of N-cadherin protein was up-regulated, but the difference was not statistically significant (P>0.05); the main pathways enriched in the differential genes of the full-length transcriptome were the PI3K-AKT and MAPK signaling pathways. CONCLUSIONS: During NSCLC development, the COX-2/PGE2/EP4 axis may promote tumor progression by inducing macrophage functional activation, and EP4 may be a potential new target for tumor immunotherapy. This study provides new perspectives and ideas for in-depth exploration of the mechanisms of NSCLC development, as well as a theoretical basis for the development of new therapeutic strategies for NSCLC.

Identification of molecular subtypes and a prognostic signature based on m6A/m5C/m1A-related genes in lung adenocarcinoma.

Lung cancer, specifically the histological subtype lung adenocarcinoma (LUAD), has the highest global occurrence and fatality rate. Extensive research has indicated that RNA alterations encompassing m6A, m5C, and m1A contribute actively to tumorigenesis, drug resistance, and immunotherapy responses in LUAD. Nevertheless, the absence of a dependable predictive model based on m6A/m5C/m1A-associated genes hinders accurately predicting the prognosis of patients diagnosed with LUAD. In this study, we collected patient data from The Cancer Genome Atlas (TCGA) and identified genes related to m6A/m5C/m1A modifications using the GeneCards database. The "ConsensusClusterPlus" R package was used to produce molecular subtypes by utilizing genes relevant to m6A/m5C/m1A identified through differential expression and univariate Cox analyses. An independent prognostic factor was identified by constructing a prognostic signature comprising six genes (SNHG12, PABPC1, IGF2BP1, FOXM1, CBFA2T3, and CASC8). Poor overall survival and elevated expression of human leukocyte antigens and immune checkpoints were correlated with higher risk scores. We examined the associations between the sets of genes regulated by m6A/m5C/m1A and the risk model, as well as the immune cell infiltration, using algorithms such as ESTIMATE, CIBERSORT, TIMER, ssGSEA, and exclusion (TIDE). Moreover, we compared tumor stemness indices (TSIs) by considering the molecular subtypes related to m6A/m5C/m1A and risk signatures. Analyses were performed based on the risk signature, including stratification, somatic mutation analysis, nomogram construction, chemotherapeutic response prediction, and small-molecule drug prediction. In summary, we developed a prognostic signature consisting of six genes that have the potential for prognostication in patients with LUAD and the design of personalized treatments that could provide new versions of personalized management for these patients.

Case of clear-cell oncocytoma of parotid gland and literature review.

Oncocytoma is a benign tumor of the salivary gland. Its incidence is very low and very seldom documen-ted in literature. Clear-cell dominant oncocytoma is even less common. The tumor's clinical symptoms and imaging results are nonspecific, so distinguishing other salivary gland tumors (such as oncocytic carcinoma) from clear-cell renal carcinoma is difficult, possibly leading to misdiagnosis and maltreatment. Here, a case of clear-cell dominant oncocytoma was presented, and the relevant literature was evaluated to investigate the diagnosis and management of clear-cell dominant oncocytoma.

Structural Priors Guided Network for the Corneal Endothelial Cell Segmentation.

The segmentation of blurred cell boundaries in cornea endothelium microscope images is challenging, which affects the clinical parameter estimation accuracy. Existing deep learning methods only consider pixel-wise classification accuracy and lack of utilization of cell structure knowledge. Therefore, the segmentation of the blurred cell boundary is discontinuous. This paper proposes a structural prior guided network (SPG-Net) for corneal endothelium cell segmentation. We first employ a hybrid transformer convolution backbone to capture more global context. Then, we use Feature Enhancement (FE) module to improve the representation ability of features and Local Affinity-based Feature Fusion (LAFF) module to propagate structural information among hierarchical features. Finally, we introduce the joint loss based on cross entropy and structure similarity index measure (SSIM) to supervise the training process under pixel and structure levels. We compare the SPG-Net with various state-of-the-art methods on four corneal endothelial datasets. The experiment results suggest that the SPG-Net can alleviate the problem of discontinuous cell boundary segmentation and balance the pixel-wise accuracy and structure preservation. We also evaluate the agreement of parameter estimation between ground truth and the prediction of SPG-Net. The statistical analysis results show a good agreement and correlation.

N,S-Doped hollow carbon nanosheet-encapsulated Co9S8 nanoparticles as a highly efficient bifunctional electrocatalyst for rechargeable zinc-air batteries.

The development of bifunctional electrocatalysts based on non-noble metals for the oxygen reduction/evolution reactions (ORR/OER) that have rationally designed structures and inexpensive components is of practical significance for the commercialization of rechargeable zinc-air batteries. Here, we report the rational synthesis of Co9S8 nanoparticles embedded in N,S co-doped hollow carbon nanosheets (Co9S8/NSC) as highly efficient oxygen electrocatalysts. The catalyst is formed when a Co-based zeolitic imidazolate framework (ZIF-67), grown on a Zn-based ZIF (ZIF-8) template, is partially vulcanized following thioacetamide (TAA) and thermal treatment. The resulting catalyst, Co9S8/NSC-3, shows satisfactory bifunctional electrocatalytic activity in 0.1 M KOH, in which the half-wave potential (E1/2) for the ORR is 0.82 V and the overpotential for the OER at 10 mA cm-2 is just 350 mV. Furthermore, as the air electrode material in a practical demonstration of a rechargeable liquid zinc-air battery, Co9S8/NSC-3 exhibits promising battery performance with a high specific capacity of 804 mA h g-1 and a pleasing charge/discharge cyclability of over 140 h at 10 mA cm-2. The satisfactory activity of Co9S8/NSC-3 can be attributed to the synergistic effect of the Co9S8 nanoparticles with the N,S-doped hollow carbon nanosheet structure, resulting in an effective electrochemically active surface with fully exposed active sites that give fast catalytic kinetics.

ZnS modified N, S dual-doped interconnected porous carbon derived from dye sludge waste as high-efficient ORR/OER catalyst for rechargeable zinc-air battery.

Facile and rational design of high-efficient oxygen reduction reaction (ORR)/oxygen evolution reaction (OER) bifunctional electrocatalysts is significant for rechargeable Zinc-air batteries. In this study, ZnS modified N, S dual-doped interconnected porous carbon (ZnS/NSC) derived from the dye sludge waste is successfully fabricated via a facile ZnCl2-assisted pyrolysis process. The effect of ZnCl2 and carbonization temperature on the microstructure and electrocatalytic performance is systematically investigated. By virtue of the synergistic effect between ZnS nanoparticles and N, S dual-doped porous carbon network, the obtained catalyst ZnS/NSC calcined at 1000 °C exhibits a decent bifunctional electrocatalytic performance with potential gap (ΔE=EOER,10-EORR,1/2) of 0.76 V comparable with commercial electrocatalysts (Pt/C and RuO2). In addition, a rechargeable zinc-air battery employed ZnS/NSC-1000 as the air cathode also displays the favorable electrochemical performance, in which the power density is 125 mW cm-2, the specific capacity is 763.27 mAh g-1 and the cycling stability at 10 mA cm-2 is more than 85 h, indicating a promising application prospect in rechargeable Zinc-air batteries.

Stress Granules Modulate SYK to Cause Tau-Associated Neurocognitive Deterioration in 5XFAD Mouse After Anesthesia and Surgery.

BACKGROUND: Alzheimer's disease (AD) is the most common type of dementia. However, no curative therapy has been found effective to slow down the process of AD. It is reported that anesthesia and surgery will induce neurocognitive deterioration in AD, but the mechanism is not quite clear. In this study, we aim to compare the cognitive impairment between 5XFAD transgenic (Tg) mice and its littermate (LM) after isoflurane anesthesia and surgery to clarify the specific impacts of anesthesia and surgery on individuals with AD and to explore the mechanisms. METHODS: We performed abdominal surgery in cognitively impaired, 4-month-old female 5XFAD mice and LM control mice. Isoflurane anesthesia (1.4%) was induced and maintained over 2 h. Open field and fear conditioning tests were conducted on 1, 3 and 7 days after anesthesia and surgery. The total distance, velocity and freezing time were the major outcomes. P-tau (AT8), tau oligomers (T22), stress granules (SGs), the SYK tyrosine kinase and p-SYK in the hippocampus at postoperative day 1 were evaluated by Western Blot assays. The colocalization of SGs, SYK, p-SYK, and neurons in the hippocampus section was assessed using qualitative immunofluorescence. RESULTS: In the open field test, no difference between the distance moved and the velocity of LM mice and 5XFAD Tg mice were found on day 1 after anesthesia and surgery. 5XFAD Tg mice exhibited reduced freezing time of fear conditioning context test on postoperative day 3, but not on day 7; the LM mice showed no changes in FCTs. Furthermore, p-tau, tau oligomers, SGs, SYK and p-SYK were evident in the hippocampus region of 5XFAD Tg mice on a postoperative day 1. In addition, SGs, SYK, p-SYK were colocalized with hippocampus neurons, as shown by immunofluorescence. CONCLUSION: This study demonstrates that anesthesia and surgery may induce tau-associated neurocognitive deterioration in individuals with AD. The mechanism under it may be associated with SGs and the tyrosine kinase, SYK. After anesthesia and surgery, in 5XFAD Tg mice, SGs were formed and SYK was phosphorylated, which may contribute to the phosphorylation of tau protein. This study provided hints that individuals with AD may be more vulnerable to anesthesia and surgery.

Amarogentin Inhibits Liver Cancer Cell Angiogenesis after Insufficient Radiofrequency Ablation via Affecting Stemness and the p53-Dependent VEGFA/Dll4/Notch1 Pathway.

BACKGROUND: Whether and how amarogentin suppresses the angiogenesis effect in liver cancer cells after insufficient radiofrequency ablation (iRFA) are still poorly studied. METHODS: The number of liver cancer stem cells (LCSCs) and the level of vascular endothelial growth factor A (VEGFA) were assessed in liver cancer tissue after iRFA. Then, CD133-positive cells were detected in iRFA models of HepG2 and Huh7 cell lines treated with amarogentin. Tube formation assays were applied to observe the antiangiogenesis effects of amarogentin. In addition, the angiogenesis-related molecules p53, delta-like ligand 4 (Dll4), and Notch1 were detected in the iRFA cells and mouse models treated with amarogentin. RESULTS: The mRNA and protein expression levels of CD133 and VEGFA were significantly higher in the residual liver cancer tissue than in the liver cancer tissues treated by hepatectomy. Amarogentin then markedly decreased the percentage of CD133-positive cells in the iRFA model in both HepG2 and Huh7 cell lines. The number of tubules formed by human umbilical vein endothelial cells (HUVECs) was significantly decreased by amarogentin. Inversely, the antiangiogenesis effect of amarogentin was counteracted after p53 silencing in the iRFA cell models. CONCLUSION: Amarogentin prevents the malignant transformation of liver cancer after iRFA via affecting stemness and the p53-dependent VEGFA/Dll4/Notch1 pathway to inhibit cancer cell angiogenesis.

Glycyrrhetinic acid alleviates hepatic inflammation injury in viral hepatitis disease via a HMGB1-TLR4 signaling pathway.

Various human disorders are cured by the use of licorice, a key ingredient of herbal remedies. Glycyrrhizic acid (GL), a triterpenoid glycoside, is the aqueous extract from licorice root. Glycyrrhetinic acid (GA) has been reported to be a major bioactive hydrolysis product of GL and has been regarded as an anti-inflammatory agent for the treatment of a variety of inflammatory diseases, including hepatitis. However, the mechanism by which GA inhibits viral hepatic inflammatory injury is not completely understood. In this study, we found that, by consecutively treating mice with a traditional herbal recipe, licorice plays an important role in the detoxification of mice. We also employed a murine hepatitis virus (MHV) infection model to illustrate that GA treatment inhibited activation of hepatic inflammatory responses by blocking high-mobility group box 1 (HMGB1) cytokine activity. Furthermore, decreased HMGB1 levels and downstream signaling triggered by injection of a neutralizing HMGB1 antibody or TLR4 gene deficiency, also significantly protected against MHV-induced severe hepatic injury. Thus, our findings characterize GA as a hepatoprotective therapy agent in hepatic infectious disease not only by suppressing HMGB1 release and blocking HMGB1 cytokine activity, but also via an underlying viral-induced HMGB1-TLR4 immunological regulation axis that occurs during the cytokine storm. The present study provides a new therapy strategy for the treatment of acute viral hepatitis in the clinical setting.

Flocculant-Assisted Synthesis of Graphene-Like Carbon Nanosheets for Oxygen Reduction Reaction and Supercapacitor.

The rational treatment of hazardous textile sludge is critical and challenging for the environment and a sustainable future. Here, a water-soluble chitosan derivative was synthesized and used as an effective flocculant in removal of reactive dye from aqueous solution. Employing these chitosan-containing textile sludges as precursors, graphene-like carbon nanosheets were synthesized through simple one-step carbonization with the use of Fe (III) salt as graphitization catalyst. It was found that the resultant graphene-like carbon nanosheets material at thickness near 3.2 nm (NSC-Fe-2) showed a high graphitization degree, high specific surface area, and excellent bifunctional electrochemical performance. As-prepared NSC-Fe-2 catalyst exhibited excellent oxygen reduction reaction (ORR) activity (onset potential 1.05 V) and a much better methanol tolerance than that of commercial Pt/C (onset potential 0.98 V) in an alkaline medium. Additionally, as electrode materials for supercapacitors, NSC-Fe-2 also displayed an outstanding specific capacitance of 195 F g-1 at 1 A g-1 and superior cycling stability (loss of 3.4% after 2500 cycles). The good electrochemical properties of the as-prepared NSC-Fe materials could be attributed to the ultrathin graphene-like nanosheets structure and synergistic effects from codoping of iron and nitrogen. This work develops a simple but effective strategy for direct conversion of textile sewage sludge to value-added graphene-like carbon, which is considered as a promising alternative to fulfill the requirements of environment and energy.

Asymmetric Polymersomes from an Oil-in-Oil Emulsion: A Computer Simulation Study.

Asymmetric vesicles are valuable for understanding and mimicking cell and practical biomedicine applications. Recently, a very straightforward methodology for fabricating asymmetric polymersome was developed by Lodge's group through the coassembly of polystyrene-b-poly(ethylene oxide) (PS-b-PEO) and polybutadiene-b-poly(ethylene oxide) (PB-b-PEO) block copolymers at the interface of a polystyrene/polybutadiene/chloroform (PS/PB/CHCl3) emulsion. However, the in-depth microscopic mechanism for the formation of asymmetric polymersomes remains unclear. To address this issue, in this article, the coassembly process for the formation of the asymmetric polymersomes in Asano's experimental system was systematically investigated by employing a dissipative particle dynamics (DPD) simulation. Our results definitely demonstrate the formation of the asymmetric polymersomes such as that in the experiments and that the bilayer formed through the folding and crossing of the PEO blocks. Besides, from the microscopic view, three stages can be discerned in the formation process: (1) the formation of micelles, (2) the micelle diffusion to the interface, and (3) the micelle rearrangement at the interface to form an asymmetric polymersome. Meanwhile, the incompatibility among PS, PB, and PEO is proven to be the main driving force for asymmetric polymersome formation. Moreover, the effects of the order of addition of copolymers and the volume fraction of PEO blocks on the structure of the asymmetric polymersomes are also investigated. We find that the formation process is affected severely by the order of addition, and adding PS-b-PEO first can make the asymmetric bilayer more perfect. Not only that, but perfect asymmetric polymersomes can be formed only when the volume fraction of PEO (fPEO) is greater than 0.55. We believe the present work can extend the knowledge of the self-assembly of asymmetric polymersomes, especially with respect to the self-assembly mechanism.

Amarogentin Induces Apoptosis of Liver Cancer Cells via Upregulation of p53 and Downregulation of Human Telomerase Reverse Transcriptase in Mice.

BACKGROUND AND OBJECTIVE: Amarogentin has been reported to have a preventive effect on liver cancer via inducing cancer cell apoptosis. We attempted to elucidate the roles of p53-associated apoptosis pathways in the chemopreventive mechanism of amarogentin. The findings of this study will facilitate the development of a novel supplementary strategy for the treatment of liver cancer. MATERIALS AND METHODS: The purity of amarogentin was assessed by high-performance liquid chromatography. The inhibitory ratios of the liver cell lines were determined using a Cell Counting Kit-8 following treatment with a gradient concentration of amarogentin. Cell apoptosis was detected by flow cytometry using annexin V-fluorescein isothiocyanate/propidium iodide kits. The gene and protein expression of p53-associated molecules, such as Akt, human telomerase reverse transcriptase, RelA, and p38, was detected by real-time quantitative polymerase chain reaction, Western blotting, and immunohistochemical staining in liver cancer cells and mouse tumor tissues after treatment with amarogentin. RESULTS: The inhibitory effect of amarogentin on cell proliferation was more obvious in liver cancer cells, and amarogentin was more likely to induce the apoptosis of liver cancer cells than that of normal liver cells. The gene and protein expression levels of Akt, RelA, and human telomerase reverse transcriptase were markedly higher in the control group than in the preventive group and treatment groups. Only the expression of human telomerase reverse transcriptase was downregulated, accompanied by the upregulation of p53. CONCLUSION: The results of our study suggest that amarogentin promotes apoptosis of liver cancer cells by the upregulation of p53 and downregulation of human telomerase reverse transcriptase and prevents the malignant transformation of these cells.

Choledochoduodenal fistula in Mainland China: a review of epidemiology, etiology, diagnosis and management.

PURPOSE: Choledochoduodenal fistula (CDF) is an extremely rare condition even in the most populous nations. However, diagnostic tools are inadequate for the young surgeon to be made aware of such a rare condition before surgery. Hence, basic understanding of the epidemiology, etiology, and management for this unusual but discoverable condition are necessary and essential. METHODS: The exclusive case reports of CDF, which were published from 1983 to 2014 concerning mainland Chinese people, were performed to review the epidemiology, etiology, and management. RESULTS: A total of 728 cases were incorporated into this review among 48 papers. More than half of the CDF cases were female (416) with an average age of 57.3 years. CDF was usually caused by cholelithiasis (573 of 728). Epigastric pain (589 of 728) and cholangitis (395 of 728) were the most common symptoms of CDF. CDF was usually detected and confirmed by endoscopic retrograde cholangiopancreatography (ERCP) (475 of 728) in Mainland China. The fistulas larger than 1 cm (82 of 654) were recommended for surgical biliary reconstruction. Fistulas between 0.5 cm and 1.0 cm (467 of 654) which were followed frequently by cholangitis attacks also required surgery; the rest were recommended to have stone removal and/or the application of an effective biliary drainage. Fistulas less than 0.5 cm (105 of 654) were usually received conservative therapy. CONCLUSION: CDF should be considered in differential diagnosis of recurrent epigastric pain and cholangitis. A possible ERCP should be arranged to investigate carefully. Depending on the size of fistula and clinical presentation, different programs for CDF are indicated, ranging from drug therapy to choledochojejunostomy.

Application of radiofrequency ablation for splenic preservation.

BACKGROUND: Traditional splenic preservation surgery involves considerable difficulties, high risks, and numerous postoperative complications. In this study, we applied radiofrequency ablation (RFA) to splenic preservation and explored its clinical value. METHODS: The clinical data of 129 patients with traumatic splenic rupture who received surgery in our hospital from September 2008-June 2013 were retrospectively analyzed. According to the operation methods, these patients were divided into three groups: 35 patients received splenic preservation surgery with RFA-assisted technique (RFA + suture repair group), 78 patients received splenic preservation surgery without RFA (traditional operation group), and 46 patients received splenectomy (splenectomy group). Preoperative, intraoperative, and postoperative-related parameters of the former two groups were compared. The postoperative complications and immunologic parameters of patients with preserved spleens were compared with those of patients who underwent splenectomy. RESULT: In the RFA + suture repair group, 34 cases successfully underwent splenic preservation surgery. Meanwhile, 49 cases successfully underwent spleen preservation surgery in the traditional operation group. RFA + suture repair group had shorter mean operation time (79 ± 22 versus 119 ± 26 min, P < 0.05), less bleeding during surgery (115 ± 67 versus 235 ± 155 mL, P < 0.05), and less intraoperative transfusion (14% versus 36%, P < 0.05). The postoperative bleeding and hospital-stay duration were remarkably lower than those in the traditional operation group (100 ± 52 versus 219 ± 93 mL and 7.1 ± 1.4 d versus 11.7 ± 2.8 d, respectively, P < 0.05). The spleen-preserving patients showed better results than the splenectomy group did for some parameters related to complications and immunology. CONCLUSIONS: Compared with traditional splenic preservation, RFA is simple and feasible, and it can greatly benefit the spleen preservation operation.

Foxp3 overexpression decreases sensitivity to chemotherapy in mouse Lewis lung cancer cells.

Chemotherapy is the main strategy for the treatment of lung cancer. However, sensitivity to chemotherapy, one of the main factors affecting the survival rate of patients with lung cancer, is extremely poor. Forkhead box P3 (Foxp3) is the key regulatory gene for the development and function of CD4+CD25+ regulatory T cells (Tregs). Increased levels of Tregs and Foxp3 expression in the peripheral blood and tumour specimens of cancer patients are associated with tumour progression and poor prognosis. In addition, certain studies have suggested that Tregs may be resistant to conventional chemotherapy and thus, enhance tumour immune evasion. Previous studies have demonstrated that Foxp3 is also expressed within tumour cells and that it may mimic the function of Tregs. Currently, the correlation between the tumour cell expression of Foxp3 and sensitivity to chemotherapy is unclear. Therefore, it was hypothesised that Foxp3 causes resistance to chemotherapeutic agents in lung cancer cells and that it may consequently promote the progression of lung cancer. In the current study, the expression of Foxp3 in mouse Lewis lung cancer (LLC) cells was detected using RT-PCR and immunocytochemistry. The overexpression of Foxp3, which was accomplished by the transient transfection of recombinant pcDNA3.1-Foxp3 or empty plasmids into LLC cells, was confirmed by RT-PCR and western blot analysis. The inhibition of cell proliferation was measured using MTT assay. The expression of multidrug resistance protein 1 (mdr1) mRNA and its protein product, P-glycoprotein (P-gp), were detected by RT-PCR and flow cytometry, respectively. The results revealed that Foxp3 was expressed by LLC cells. The inhibitory rate of cell proliferation in Foxp3-overexpressing LLC cells compared with those transfected with an empty plasmid was significantly decreased following adriamycin (ADM) and mitomycin C (MMC) treatment. The IC50 values of ADM and MMC in Foxp3-overexpressing LLC cells were increased. The expression levels of mdr1 mRNA and P-gp were significantly upregulated in Foxp3 overexpressing LLC cells. These results suggest that Foxp3 reduces the sensitivity of LLC cells to ADM and MMC, thus promoting tumour progression, by upregulating the expression of mdr1 mRNA and P-gp.

Foxp3 expression in A549 cells is regulated by Toll-like receptor 4 through nuclear factor-κB.

Foxp3 is a master regulator of the development and function of CD4+CD25+ regulatory T cells (Tregs). Previous studies have reported that Foxp3 is also expressed in tumour cells and promotes tumour immune evasion. However, the regulation of the expression of Foxp3 in tumour cells remains unclear. Toll-like receptor 4 (TLR4), a member of the pattern recognition receptor family, is also expressed in tumour cells. Previous studies have found that the TLR4 signaling pathway is involved in tumour immune evasion in lung cancer cells, and that the transcription factor, nuclear factor-κB (NF-κB), plays a key role in the TLR4 signaling pathway. Moreover, recent studies found that NF-κB promotes the transcription of Foxp3. We hypothesised that TLR4 may also be involved in the regulation of Foxp3 in A549 cells through NF-κB. Therefore, we examined the effect of TLR4 and NF-κB on the expression of Foxp3 in the A549 lung cancer cell line. The results showed that Foxp3 and TLR4 are expressed in A549 cells; the expression of Foxp3 increased after lipopolysaccharide (LPS) stimulation and decreased after blocking the TLR4 signaling pathway. In addition, the expression of NF-κB (p65) increased after LPS stimulation and the expression of Foxp3 decreased after blocking NF-κB. These results suggest that TLR4 is involved in the regulation of Foxp3 in A549 cells through NF-κB.

Long-term immunogenicity after single and booster dose of a live attenuated hepatitis A vaccine: results from 8-year follow-up.

Live, attenuated hepatitis A vaccines are used widely in China but there is uncertainty regarding the persistence of vaccine-induced anti-HAV antibodies after single dose and booster dose administrated at month 12. A large scale clinical trial to evaluate the live, attenuated hepatitis A vaccine was conducted in Hebei province between 1996 and 1999. Five years after the trials, children in single dose and booster dose groups were bled and followed. Seventy two percent (61/85) of children who received a single trial dose had detectable anti-HAV antibodies for 96 months (GMC at 96 months: 89.0 mIU/mL). In the booster group 98% (48/49) children remained anti-HAV positive with GMC of 262.8 mIU/mL at month 96. The reinjection with live attenuated HAV vaccine can elicit a booster effect. Results from single dose group seems not to support the need for booster doses of live attenuated hepatitis A vaccine in immunocompetent individuals regarding the persisting anti-HAV and anamnestic response of a single dose vaccine. Continued monitoring of anti-HAV antibodies is needed for a rational hepatitis A immunization strategy in China.