Publications by authors named "Chengfeng Xu"

16 Publications

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Comprehensive analysis of tumor microenvironment and identification of an immune signature to predict the prognosis and immunotherapeutic response in lung squamous cell carcinoma.

Ann Transl Med 2021 Apr;9(7):569

Department of Thoracic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.

Background: Tumor mutation burden (TMB) and immune microenvironment are important determinants of prognosis and immunotherapeutic efficacy for cancer patients. The aim of the present study was to develop an immune signature to effectively predict prognosis and immunotherapeutic response in patients with lung squamous cell carcinoma (LUSC).

Methods: TMB and immune microenvironment characteristics were comprehensively analyzed by multi-omics data in LUSC. The immune signature was further constructed and validated in multiple independent datasets by LASSO Cox regression analysis. Next, the value of immune signature in predicting the response of immunotherapy was evaluated. Finally, the possible mechanism of immune signature was also investigated.

Results: A novel immune signature based on 5 genes was constructed and validated to predict the prognosis of LUSC patients. These genes were filamin-C, Rho family GTPase 1, interleukin 4-induced gene-1, transglutaminase 2, and prostaglandin I2 synthase. High-risk patients had significantly poorer survival than low-risk patients. A nomogram was also developed based on the immune signature and tumor stage, which showed good application. Furthermore, we found that the immune signature had a significant correlation with immune checkpoint, microsatellite instability, tumor infiltrating lymphocytes, cytotoxic activity scores, and T-cell-inflamed score, suggesting low-risk patients are more likely to benefit from immunotherapy. Finally, functional enrichment and pathway analyses revealed several significantly enriched immune-related biological processes and metabolic pathways.

Conclusions: In the present study, we developed a novel immune signature that could predict prognosis and immunotherapeutic response in LUSC patients. The results not only help identify LUSC patients with poor survival, but also increase our understanding of the immune microenvironment and immunotherapy in LUSC.
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http://dx.doi.org/10.21037/atm-21-463DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8105790PMC
April 2021

Identification and characterization of hirudin-HN, a new thrombin inhibitor, from the salivary glands of .

PeerJ 2019 30;7:e7716. Epub 2019 Sep 30.

Institute of Chinese Medicinal Materials, Nanjing Agricultural University, Nanjing, China.

Transcriptome sequencing data (6.5 Gb) of the salivary glands of the haematophagous leech was obtained by using the BGIseq-500 platform. After identification and analysis, one transcript (Unigene5370) was annotated to hirudin HV3 from with an -value of 1e-29 and was named hirudin-HN. This transcript was a new thrombin inhibitor gene belonging to the proteinase inhibitor I14 (hirudin) family. Hirudin-HN, with a 270-bp cDNA, encodes an 89-aa protein containing a 20-aa signal peptide. The mature hirudin-HN protein contains the typical structural characteristics of hirudin, e.g., three conserved disulfide bonds and the PKP and DFxxIP motifs. Proteins ( and ) were obtained via prokaryotic expression, and the mature hirudin-HN protein was shown to have anticoagulant activity and thrombin affinity by using the chromogenic substrate S2238 and surface plasmon resonance (SPR) interaction analysis, respectively. The N-terminal structure of the mature hirudin-HN protein was shown to be important for anticoagulant activity by comparing the activity and thrombin affinity of and . The abundances of Hirudin-HN mRNA and protein were higher in the salivary glands of starving animals than in those of feeding or fed leeches. These results provided a foundation for further study on the structure-function relationship of hirudin-HN with thrombin.
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http://dx.doi.org/10.7717/peerj.7716DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6776071PMC
September 2019

Species identification of Bombyx mori and Antheraea pernyi silk via immunology and proteomics.

Sci Rep 2019 06 28;9(1):9381. Epub 2019 Jun 28.

Key Laboratory of Advanced Textile Materials and Manufacturing Technology, Ministry of Education, Zhejiang Sci-Tech University, Hangzhou, 310018, China.

In recent years, increasing attention has been paid to the origin, transmission and communication of silk. However, this is still an unsolved mystery in archaeology. The identification of silk-producing species, especially silk produced by Bombyx mori (B. mori) and Antheraea pernyi (A. pernyi), is of key significance to address this challenge. In this study, two innovative methods, i.e. immunology and proteomics, were proposed and successfully established for the species identification of silks. ELISAs result demonstrated that the two prepared antibodies exhibited high sensitivity and specificity in distinguishing B. mori and A. pernyi silk. No cross-reactivity with each other was observed. Moreover, biomarkers were obtained for Bombyx and Antheraea through proteomic analysis. It was also confirmed that the biomarkers were suitable for identifying the species that produced a given silk sample. Compared with conventional methods for distinguishing silk species, immunological and proteomics techniques used in tandem can provide intact information and have the potential to provide accurate and reliable information for species identification.
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http://dx.doi.org/10.1038/s41598-019-45698-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6599025PMC
June 2019

Gene cloning and expression of a partial sequence of Hirudomacin, an antimicrobial protein that is increased in leech (Hirudo nipponica Whitman) after a blood meal.

Comp Biochem Physiol B Biochem Mol Biol 2019 May 19;231:75-86. Epub 2019 Feb 19.

Institute of Chinese Medicinal Materials, Nanjing Agricultural University, Nanjing 210095, China.

The novel antimicrobial gene Hirudomacin (Hmc), with a 249-bp cDNA, encodes a mature protein of 61 amino acids and a 22-amino acid signal peptide. Hmc exhibits the highest similarity, at 90.1%, with macin family members found in the salivary gland of the leech Hirudo nipponica Whitman. A mature Hmc protein concentration of 219 μg/mL was detected using the Bradford method. The mature Hmc protein is 6862.82 Da and contains 8 cysteine residues. Antimicrobial assays showed a minimum bactericidal concentration and 50% lethal dose of 1.56 μg/mL and 0.78 μg/mL, respectively, for Staphylococcus aureus and 0.39 μg/mL and 0.195 μg/mL, respectively, for Bacillus subtilis. Transmission electron microscopy revealed membrane integrity disruption in S. aureus and B. subtilis, which resulted in bacterial lysis. The level of Hmc mRNA in the salivary gland during three blood meal stages indicated a remarkable trend of increase (P < .05), and western blotting demonstrated that among the three blood meal stages, expression of the mature Hmc protein was highest in both the salivary gland and intestine at the fed stage (P < .05). Immunofluorescence further showed the mature Hmc protein to be localized outside the cell nucleus, with the signal intensity in the salivary gland peaking at the fed stage (P < .05). In conclusion, the mature Hmc protein exhibits broad-spectrum antimicrobial effects against gram-positive and gram-negative bacteria, and a blood meal upregulates Hmc gene and protein expression in H. nipponica.
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http://dx.doi.org/10.1016/j.cbpb.2019.02.005DOI Listing
May 2019

Species Identification of Silks from Bombyx mori, Eri Silkworm and Chestnut Silkworm Using Western Blot and Proteomics Analyses.

Anal Sci 2019 Feb 28;35(2):175-180. Epub 2018 Sep 28.

Key Laboratory of Advanced Textile Materials and Manufacturing Technology, Ministry of Education, Zhejiang Sci-Tech University.

Species identification is of key significance for exploring the origin and transmission of ancient silks. In this study, two novel methods, i.e. western blot (WB) and proteomics analyses, were proposed and established to identify the differences between silks from Bombyx mori (B. mori) and two other distinctive species (Eri silkworm and Chestnut silkworm). Three diagnostic antibodies, a polyclonal anti-silk fibroin (anti-SF) antibody (pAb), a polyclonal anti-SF-specific peptide antibody (pAsb), and a monoclonal anti-SF antibody (mAb) were designed and prepared to distinguish silk species using the antibody-based WB technique. Proteomics analysis by liquid chromatography-tandem mass spectrometry was performed to further identify silk species at the protein level. WB results indicated that the three antibodies showed high specificity and affinity and could discern B. mori silk from Eri and Chestnut silks. Biomarkers for each SF were obtained using proteomics analysis, and they have the potential to serve as standards for identifying silk species. Thus, combining WB and proteomics analyses with conventional methods can provide more accurate silk information and may be suitable for identifying other proteinaceous materials in archaeological field.
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http://dx.doi.org/10.2116/analsci.18P314DOI Listing
February 2019

Psoralen induced cell cycle arrest by modulating Wnt/β-catenin pathway in breast cancer cells.

Sci Rep 2018 09 18;8(1):14001. Epub 2018 Sep 18.

Department of Thyroid and Breast Surgery, Binzhou Medical University Hospital, Binzhou, Shandong, 256603, P. R. China.

Psoralen could inhibit the proliferation of human breast cancer cells, however, the molecular mechanism was unclear. We evaluated the anti-proliferative effects of psoralen by MTT, plate colony formation assay and cell cycle analysis in MCF-7 and MDA-MB-231 cells. The effects of psoralen on activation of Wnt/β-catenin and the related target genes were examined by quantitative real-time PCR, western blotting and cell immunofluorescence. The tumor growth was conducted in BALB/c nude mice and the pathological changes of heart, liver and kidney were also observed. Our results demonstrate that psoralen significantly inhibited cell proliferation by inducing G0/G1 phase arrest in MCF-7 cells and G2/M phase arrest in MDA-MB-231 cells. The expression of Fra-1 was reduced and Axin2 was promoted both in MCF-7 and MDA-MB-231 cells after psoralen treatment. The cytoplasmic accumulation and nuclear translocation of β-catenin were significantly reduced by psoralen. Psoralen increased the levels of phospho-(Y142) β-catenin, while decreased the expression of total β-catenin and its downstream target Fra-1 in vitro and vivo. Moreover, psoralen didn't cause any significant toxicity at the effective concentration. Overall, our results might provide theoretical basis for clinical application of psoralen in breast cancer.
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http://dx.doi.org/10.1038/s41598-018-32438-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6143618PMC
September 2018

Development of an Enzyme-Linked Immunosorbent Assay and Gold-Labelled Immunochromatographic Strip Assay for the Detection of Ancient Wool.

J Anal Methods Chem 2018 5;2018:2641624. Epub 2018 Jun 5.

Institute of Textile Conservation, Zhejiang Sci-Tech University, Hangzhou 310018, China.

The identification of ancient wool is of great importance in archaeology. Despite lots of meaningful information can be achieved by conventional detection methods, that is, light and electron microscopy, spectroscopy, and chromatography, the efficacy is likely to be limited in the detection of ancient samples with contamination or severe degradation. In this work, an immunoassay was proposed and performed for the identification of ancient wool. First, a specific antibody, which has the benefits of low cost, easy operation, and extensive applicability, was developed directly through immunizing rabbits with complete antigen (keratin). Then, an enzyme-linked immunosorbent assay (ELISA) and a colloidal gold-labelled immunochromatographic strip (ICS) were developed to qualitatively identify the corresponding protein in ancient wool samples unearthed from Kazakhstan and China. The anti-keratin antibody exhibited high sensitivity and specificity for the identification of modern and ancient wool. The limit of detection (LOD) of the ELISA method was 10 ng/mL, and no cross-reactions with other interfering antigens have been noted. It is concluded that the immunoassays are reliable methods for the identification of ancient wool.
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http://dx.doi.org/10.1155/2018/2641624DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6008753PMC
June 2018

Exosomes play an important role in the process of psoralen reverse multidrug resistance of breast cancer.

J Exp Clin Cancer Res 2016 12 1;35(1):186. Epub 2016 Dec 1.

Department of Thyroid and Breast Surgery, The Affiliated Hospital of Binzhou Medical University, 522 Yellow Three Road, Binzhou, Shandong, 256603, People's Republic of China.

Background: Release of exosomes have been shown to play critical roles in drug resistance by delivering cargo. Targeting the transfer of exosomes from resistant cells to sensitive cells may be an approach to overcome some cases of drug resistance.

Method: In this study, we investigated the potential role of exosomes in the process of psoralen reverse multidrug resistance of MCF-7/ADR cells. Exosomes were isolated by differential centrifugation of culture media from MCF-7/ADR cells (ADR/exo) and MCF-7 parental cells (S/exo). Exosomes were characterized by morphology, exosomal markers and size distribution. The ability of ADR/exo to transfer multidrug resistance was assessed by MTT and real-time quantitative PCR. The different formation and secretion of exosomes were detected by immunofluorescence and transmission electron microscopy. Then we performed comparative transcriptomic analysis using RNA-Seq technology and real-time quantitative PCR to better understand the gene expression regulation in exosmes formation and release after psoralen treatment.

Results: Our data showed that exosomes derived from MCF-7/ADR cells were able to promote active sequestration of drugs and could induce a drug resistance phenotype by transferring drug-resistance-related gene MDR-1 and P-glycoprotein protein. Psoralen could reduce the formation and secretion of exosomes to overcome drug resistance. There were 21 differentially expressed genes. Gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most significantly expressed genes were linked to PPAR and P53 signaling pathways which were related to exosomes formation, secretion and cargo sorting.

Conclusions: Psoralen can affect the exosomes and induce the reduction of resistance transmission via exosomes might through PPAR and P53 signaling pathways, which might provide a novel strategy for breast cancer resistance to chemotherapy in the future.
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http://dx.doi.org/10.1186/s13046-016-0468-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5131502PMC
December 2016

Psoralen reverses the P-glycoprotein-mediated multidrug resistance in human breast cancer MCF-7/ADR cells.

Mol Med Rep 2016 Jun 8;13(6):4745-50. Epub 2016 Apr 8.

Department of Thyroid and Breast Surgery, The Affiliated Hospital of Binzhou University of Medicine, Binzhou, Shandong 256603, P.R. China.

The resistance of cancer to chemotherapeutic agents is a major obstacle during chemotherapy. Clinical multidrug resistance (MDR) is commonly mediated by membrane drug efflux pumps, including ATP‑binding cassette subfamily B member 1, also termed P-glycoprotein (P-gp). P-gp is a membrane transporter encoded by the MDR1 gene. The current study aimed to investigate the impact of psoralen on the expression and function of P‑gp. The 10% inhibitory concentration (IC10) of psoralen, and its capacity to reduce MDR in adriamycin (ADR)‑resistant MCF‑7/ADR cells were determined using MTT assay. The ability of psoralen to modulate the transport activity of P‑gp in MCF‑7/ADR cells was evaluated by measuring the accumulation and efflux of rhodamine 123 (Rh 123) and adriamycin with flow cytometry. The present study evaluated the mRNA level of MDR1 in MCF‑7 and MCF‑7/ADR cells treated with psoralen using reverse transcription-quantitative polymerase chain reaction. The protein expression level of P‑gp was examined by western blot analysis. The current study demonstrated that the IC10 of psoralen in MCF‑7/ADR cells was 8 µg/ml. At 8 µg/ml, psoralen reduced MDR and the sensitivity of the MCF‑7/ADR cells to ADR compared with untreated cells. Additionally, psoralen significantly increased the intracellular accumulation of ADR and Rh 123. However, the IC10 of psoralen did not affect the protein expression levels of P‑gp or mRNA levels of MDR1 (P>0.05). Psoralen reduces MDR by inhibiting the efflux function of P‑gp, which may be important for increasing the efficiency of chemotherapy and improving the clinical protocols aiming to reverse P-gp-mediated MDR.
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http://dx.doi.org/10.3892/mmr.2016.5098DOI Listing
June 2016

The extracellular matrix protein matrilin-2 induces post-burn inflammatory responses as an endogenous danger signal.

Inflamm Res 2015 Oct 14;64(10):833-9. Epub 2015 Aug 14.

Department of Burn and Plastic Surgery, The First Affiliated Hospital of PLA General Hospital, 51 Fucheng Road, Beijing, 100048, China.

Objective And Design: This prospective experimental study aims to investigate whether matrilin-2 is released from burn injury and induces post-burn inflammatory responses as an endogenous danger signal.

Subjects: Fifteen burn patients, 15 volunteers, 12 matrilin-2-deficient mice, 36 C57BL/6 mice and raw 264.7 cells.

Methods: Matrilin-2 levels were detected by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction. The inflammatory cytokines production in Matn2 deficient mice and wide type mice were detected by ELISA. Macrophages were activated by recombinant mouse MATN2 with or without adding anti-Toll-like receptor (TLR) 4 antibody. Student's t test and one-way analysis of variance were used for statistical analysis.

Results: The matrilin-2 levels in serum of burned patients were drastically elevated as compared to those of healthy controls. The matrilin-2 levels in burned mice were significantly increased than those of non-burned controls, whereas the matrilin-2 mRNA expression was not significantly changed after burn. In addition, Matn2 deficient mice showed remarkably less inflammatory cytokines production and less neutrophil infiltration in lung. Exogenous MATN2 induced potent expression of proinflammatory cytokines production in macrophages, which was inhibited by anti-TLR4 antibody.

Conclusion: Matrilin-2 induces post-burn inflammatory responses as an endogenous danger signal, partly through a TLR4-mediated mechanism.
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http://dx.doi.org/10.1007/s00011-015-0867-0DOI Listing
October 2015

Apelin inhibits the activation of the nucleotide-binding domain and the leucine-rich, repeat-containing family, pyrin-containing 3 (NLRP3) inflammasome and ameliorates insulin resistance in severely burned rats.

Surgery 2015 Jun 25;157(6):1142-52. Epub 2015 Mar 25.

Department of Burn and Plastic Surgery, The First Affiliated Hospital of PLA General Hospital, Beijing, China.

Background: Hyperglycemia with insulin resistance remains a challenging problem in severely burned patients. Recent studies indicated the involvement of the nucleotide-binding domain and the leucine-rich, repeat-containing family, pyrin-containing 3 (NLRP3) inflammasome in insulin resistance and a beneficial role of apelin in insulin resistance. Our aim was to investigate whether apelin inhibits the activation of the NLRP3 inflammasome and ameliorates insulin resistance in severely burned rats.

Methods: Male Wistar rats were subjected to a full-thickness burn injury comprising 40% of the total body surface area and were randomized to receive apelin, N(G)-methyl-L-arginine acetate salt (L-NMMA), and apelin plus treatments with L-NMMA. The following outcome measurements were assessed: apelin/APJ mRNA expression in white adipose tissue (WAT) and muscles, plasma apelin level, and activation of the NLRP3 inflammasome in WAT, Interleukin-1 β, interleukin-6, tumor necrosis factor-α, and monocyte chemoattractant protein-1 levels in plasma, insulin resistance, survival rates, and endothelial nitric oxide synthase phosphorylation in soleus muscles.

Results: Severe burn induced a decreased expression of apelin/APJ mRNA in soleus muscles and a decrease in plasma apelin levels. Burn injury with apelin treatment restored plasma apelin level, inhibited NLRP3 inflammasome activity in WAT, and decreased inflammatory cytokine levels in plasma. Rats treated with apelin also showed improved insulin sensitivity and decreased mortality, accompanied by a remarkable induction of endothelial nitric oxide synthase phosphorylation in soleus muscle. Furthermore, the aforementioned effects of apelin were inhibited in part by treatment with L-NMMA.

Conclusion: Apelin inhibits the activation of NLRP3 inflammasome, attenuates systemic inflammatory response, ameliorates insulin resistance, and promotes survival after severe burn, in part through an endothelial nitric oxide synthase-dependent pathway.
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http://dx.doi.org/10.1016/j.surg.2015.01.011DOI Listing
June 2015

Antibiotic resistance and an in vitro biofilm model.

Burns 2012 Feb 12;38(1):141; author reply 141-2. Epub 2011 Nov 12.

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http://dx.doi.org/10.1016/j.burns.2011.03.025DOI Listing
February 2012

Bone marrow transplantation combined with mesenchymal stem cells induces immune tolerance without cytotoxic conditioning.

J Surg Res 2011 Nov 13;171(1):e123-31. Epub 2011 Jul 13.

Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi Province, China.

Background: Transplantation of allogeneic donor bone marrow (BM) into sufficiently conditioned recipients is an effective approach to induce immune tolerance as assessed by mixed chimerism. However, this approach is hampered by the lack of feasible protocols devoid of cytoreductive conditioning. We investigated whether mixed chimerism could be established by intra-bone marrow-bone marrow transplantation (IBM-BMT) combined with bone marrow-derived mesenchymal stem cells (BMSCs) treatment without additional cytoreductive conditioning.

Materials And Methods: The recipient mice (C57BL/6) accepted BMSCs from donor mice (Balb/c) through daily tail vein injection for 4 d followed by IBM-BMT immediately. Full-thickness skin grafts from donor mice as well as from the third party mice (ICR) were transplanted to the dorsum of the recipient mice after the combined IBM-BMT with BMSCs treatment. The immune tolerance was assessed by the survival time of skin allografts. The establishment of mixed chimerism and cytokine expression profile in recipient peripheral blood were determined by flow cytometry and enzyme-linked immunosorbent assay, respectively.

Results: IBM-BMT combined with BMSCs treatment led to stable mixed chimerism and donor-specific skin graft tolerance. The flow cytometry analysis revealed that recipient mice developed 20%-25% chimerism levels among the myeloid lineage. The skin allografts survived more than 1 y and the hair re-grew normally on the grafts. Cytokine profile showed that IBM-BMT combined with BMSCs treatment prolonged humoral tolerance in recipient chimeras.

Conclusions: Our results demonstrate that donor specific immune tolerance can be effectively induced by IBM-BMT combined with BMSCs treatment without any additional cytoreductive recipient treatment. This approach provides a promising allograft transplantation strategy when the donor bone marrow is available.
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http://dx.doi.org/10.1016/j.jss.2011.06.020DOI Listing
November 2011

Concerns about stability and accuracy of a burn model.

Burns 2011 Nov 6;37(7):1272; author reply 1272-3. Epub 2011 Aug 6.

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http://dx.doi.org/10.1016/j.burns.2011.06.011DOI Listing
November 2011

Aggregation properties of a novel class of amphiphilic cationic polyelectrolytes containing gemini surfactant segments.

J Phys Chem B 2010 Oct;114(39):12448-54

School of Chemical and Environmental Engineering, China University of Mining and Technology (Beijing), Beijing, 100083, People's Republic of China.

A novel class of amphiphilic cationic polyelectrolytes, poly(A-co-G)s, comprising of gemini type surfactant segment 1,3-bis(N,N-dimethyl-N-dodecylammonium)-2-propylacrylate dibromide (G) and acryloyloxyethyl trimethyl ammonium chloride (A), were synthesized. Their aggregation properties were investigated by employing fluorescence spectroscopy, dynamic light scattering, transmission electron microscopy, and ζ-potential measurements. For comparison, a series of polyelectrolytes containing a traditional single alkyl chain surfactant unit (acryloyloxyethyl-N,N-dimethyl-N-dodecylammonium bromide (D)), poly(A-co-D)s, were also synthesized and investigated. It was found that the critical aggregation concentration (cac) of poly(A-co-G)s is much lower than that of poly(A-co-D)s. The huge interpolymer aggregates (with a hydrodynamic radius of >450 nm) occur in poly(A-co-G)s aqueous solution, and the size of aggregates increases with the increase of the molar content of the gemini-type surfmer segment and the concentration of the copolymer. The size of aggregates in poly(A-co-D)s aqueous solution is much smaller than poly(A-co-G)s, which also increases with the increase of the molar content of the single alkyl chain surfmer segment and the concentration of the copolymer. The results of aggregation number and charge density of aggregate in poly(A-co-G)s and poly(A-co-D)s indicate that the copolymers have a strong tendency toward interpolymer aggregation and the aggregates in poly(A-co-G)s are much more compact than those of poly(A-co-D)s. These results are interpreted in terms of the synergistic effects of double hydrophobic chains on the gemini surfactant unit.
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http://dx.doi.org/10.1021/jp102685wDOI Listing
October 2010

Rotavirus vaccine and severe diarrhea in African infants.

N Engl J Med 2010 Apr;362(17):1637; author reply 1638

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April 2010
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