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Background: Burn wound healing is a complex physiological process that requires complicated regulation by different cells and tissues. Brown adipose tissue (BAT) plays a key role in the hypermetabolic response to severe burns. However, it is unclear whether BAT contributes to burn wound healing., Methods: Mice were divided into two groups: brown adipose tissue removal group (BR group) and control group. Burn wounds were created on the backs of mice (weighing 20-25g), who were exposed to 100°C hot water for 12 seconds using a homemade burn tube, resulting in a burned area measuring 10 mm in diameter. The treatments were applied once a day for 10 days. Full-thickness wound tissue was collected on days 1, 4, 7, 10, and analyzed by immunostaining of CD31，α-SMA+, F4/80 and CD206 (n = 3)., Results: On days 4, 7, and 10, the wound healing rate of the control group was significantly higher than that of the BR group. In the histological analysis, evident inflammatory infiltration, severe collagen denaturation in the BR group. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the IL-17 pathway was enriched and related genes were up-regulated in the heat map. Immunostaining and transcriptional analyses revealed that angiogenesis and fibroblast were enhanced in the control group, fewer CD206-positive M2 macrophages and higher levels of inflammatory infiltration in the BR group., Conclusions: Brown adipose tissue may reduce inflammatory signaling in burn wounds by increasing the IL-17A-HIF1α axis and driving M2 macrophage polarization., (Copyright © 2024 by the American Society of Plastic Surgeons.)

In the fight against oral cancer, innovative methods like Raman spectroscopy and deep learning have become powerful tools, particularly in integral tasks encompassing tumor staging, lymph node staging, and histological grading. These aspects are essential for the development of effective treatment strategies and prognostic assessment. However, it is important to note that most research so far has focused on solutions to one of these problems and has not taken full advantage of the potential wealth of information in the data. To compensate for this shortfall, we conceived a method that combines Raman spectroscopy with deep learning for simultaneous processing of multiple classification tasks, including tumor staging, lymph node staging, and histological grading. To achieve this innovative approach, we collected 1750 Raman spectra from 70 tissue samples, including normal and cancerous tissue samples from 35 patients with oral cancer. In addition, we used a deep neural network architecture to design four distinct multi-task network (MTN) models for intelligent oral cancer diagnosis, named MTN-Alexnet, MTN-Googlenet, MTN-Resnet50, and MTN-Transformer. To determine their effectiveness, we compared these multitask models to each other and to single-task models and traditional machine learning methods. The preliminary experimental results show that our multi-task network model has good performance, among which MTN-Transformer performs best. Specifically, MTN-Transformer has an accuracy of 81.5%, a precision of 82.1%, a sensitivity of 80.2%, and an F1_score of 81.1% in terms of tumor staging. In the field of lymph node staging, the accuracy, precision, sensitivity, and F1_score of MTN-Transformer are 81.3%, 83.0%, 80.1%, and 81.5% respectively. Similarly, for the histological grading classification tasks, the accuracy was 83.0%, the precision 84.3%, the sensitivity 76.7%, and the F1_score 80.2%. This code is available at https://github.com/ISCLab-Bistu/MultiTask-OralRamanSystem.

This study tried to know how self-regulation strategies (SRS) affected the motivation, self-efficacy, willingness to communicate (WTC), and creativity of Chinese EFL learners as they learned English language. To do this, eighty intermediate EFL students were selected and divided into two groups: experimental and control. Four questionnaires were then used to assess the participants' motivation for language acquisition, WTC, self-efficacy, and creativity prior to the intervention. Subsequently, SRS was used to treat the experimental group (EG), whereas a traditional instruction was given to the control group (CG) without the use of SRSs. Four questionnaires were given again as post-tests following a 15-session treatment to gauge how the treatment affected the students' motivation for language acquisition, WTC, self-efficacy, and creativity. Based on the study's results, the EG considerably outstripped the CG on each of the four post-tests. The findings of this research have some implications for researchers, English teachers, EFL learners, and other stakeholders., (© 2024. The Author(s).)

Objective: This article reports two accidents caused by defective Takata airbags ruptured, which led to the deaths of the drivers. This is the first public report on the deaths caused by Takata airbags in China., Methods: Determine the relationship between the driver death and airbag rupture through autopsy indings and vehicle inspection., Results: Due to defects in the design of Takata's inflator, moist air was permitted to slowly enter the inflator, resulting the PSAN slowly degraded physically. The damaged propellant burned more rapidly than intended and overpressurized the inflator's steel housing, causing fragmentation and flying debris at high speed, killing or injuring vehicle occupants., Conclusions: To date, there are still tens of millions of defective Takata airbags that have not been recalled for repair, posing safety risks. This article suggests taking preventive measures to avoid the occurrence of similar accidents.

Background: Immunosenescence occurs as people age, leading to an increased incidence of age-related diseases. The number of senescent T cells also rises with age. T cell senescence and immune response dysfunction can result in a decline in immune function, especially in anti-tumor immune responses. Metformin has been shown to have various beneficial effects on health, such as lowering blood sugar levels, reducing the risk of cancer development, and slowing down the aging process. However, the immunomodulatory effects of metformin on senescent T cells still need to be investigated., Methods: PBMCs isolation from different age population (n = 88); Flow Cytometry is applied to determine the phenotypic characterization of senescent T lymphocytes; intracellular staining is applied to determine the function of senescent T cells; Enzyme-Linked Immunosorbent Assay (ELISA) is employed to test the telomerase concentration. The RNA-seq analysis of gene expression associated with T cell senescence., Results: The middle-aged group had the highest proportion of senescent T cells. We found that metformin could decrease the number of CD8 + senescent T cells. Metformin affects the secretion of SASP, inhibiting the secretion of IFN-γ in CD8 + senescent T cells. Furthermore, metformin treatment restrained the production of the proinflammatory cytokine IL-6 in lymphocytes. Metformin had minimal effects on Granzyme B secretion in senescent T cells, but it promoted the production of TNF-α in senescent T cells. Additionally, metformin increased the concentration of telomerase and the frequency of undifferentiated T cells. The results of RNA-seq showed that metformin promoted the expression of genes related to stemness and telomerase activity, while inhibiting the expression of DNA damage-associated genes., Conclusion: Our findings reveal that metformin could inhibit T cell senescence in terms of cell number, effector function, telomerase content and gene expression in middle-aged individuals, which may serve as a promising approach for preventing age-related diseases in this population., (© 2023. The Author(s).)

In this report, we present a case study of a rare human bacterium, Corynebacterium bovis , which caused an infection in a patient who had undergone autologous fat-based breast augmentation using cryopreserved fat. This infection occurred during a secondary fat grafting procedure. To identify the bacteria causing the infection, we used high-throughput DNA sequencing technology since this bacterium is seldomly reported in human infections. The patient was successfully treated with intravenous imipenem. We also discuss potential factors that may have contributed to this unusual bacterial infection and propose that DNA sequencing can be a useful tool in cases where standard culture techniques fail to identify the causative agent. Additionally, we highlight the importance of further research on the cryopreservation of fat. In summary, this case highlights the possibility of rare bacterial infections occurring after fat grafting procedures and emphasizes the importance of identifying the causative agent through advanced techniques such as DNA sequencing. Further research is needed to improve our understanding of the risks associated with cryopreservation of fat and to identify ways to prevent these types of infections in the future., 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 You, Yao, Gao and Liao.)

Introduction: Bananas are not only an important food crop for developing countries but also a major trading fruit for tropical and semitropical regions, maintaining a huge trade volume. Fusarium wilt of banana (FWB) caused by Fusarium oxysporum f. sp. cubense is becoming a serious challenge to the banana industry globally. Biological control has the potential to offer both effective and sustainable measures for this soil-borne disease., Methods: In order to explore the biocontrol effects of the biological agent Bacillus amyloliquefaciens QST713 strain on banana plants, two cultivars, Brazilian and Yunjiao No. 1, with varied resistance to FWB, were used in greenhouse pot experiments., Results: Results showed that the plant height and pseudostem diameter of banana-susceptible cultivar Brazilian increased by 11.68% and 11.94%, respectively, after QST713 application, while the plant height and pseudostem diameter of resistant cultivar Yunjiao No. 1 increased by 14.87% and 12.51%, respectively. The fresh weight of the two cultivars increased by 20.66% and 36.68%, respectively, indicating that this biological agent has potential effects on plant growth. Analysis of the rhizosphere soil microbial communities of two different cultivars of banana plants showed that TR4 infection and B. amyloliquefaciens QST713 strain application significantly affected the bacterial and fungal diversity of Yunjiao No. 1, but not in the cultivar Brazilian. In addition, TR4 infection and QST713 application changed the bacterial community composition of both banana cultivars, and the fungal community composition of Yunjiao No. 1 also changed significantly. Relevance analysis indicated that the relative richness of Bacillus and Pseudomonas in the rhizosphere of both cultivars increased significantly after QST713 application, which had a good positive correlation with plant height, pseudostem girth, aboveground fresh weight, leaf length, and leaf width., Discussion: Therefore, the outcome of this study suggests that the biological agent QST713 strain has potential application in banana production for promoting plant growth and modification of soil microbial communities, particularly in the TR4-infected field., 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 Tian, Zhang, Zhou, Li, Wang, Yang, Bai, Fan, He and Zheng.)

Background and Objectives: Addictive behaviors are serious factors for mental health and usually increase during public crises. We identified the vulnerable characteristics for bad prognosis of addictive internet use across different periods of the coronavirus disease 2019 (COVID-19) pandemic., Methods: Self-reported questionnaires were delivered in three waves through jdh.com during the outbreak (n = 17,960), remission (n = 15,666), and dynamic zero (n = 12,158) periods of COVID-19 pandemic in China. Internet addiction degree was assessed using the Internet Addiction Test. The different progression groups were divided using a latent class growth model among 1679 longitudinal participants. Risk factors for bad progression were identified by two-step logistic regression., Results: A total of 40.16% of participants reported an increase in the addictive degree of internet use compared with prepandemic. Across different COVID-19 periods, the overall trend of addictive internet use was downward among general Chinese study participants (Mslope = -1.56). Childhood traumatic experiences, deterioration of physical health, depression, and anxiety during remission and dynamic periods were the main risk factors for the bad progression of pandemic-induced addictive internet use., Discussion and Conclusions: Addictive internet use was remitted following relaxed control policies during the COVID-19 pandemic. Negative childhood experiences and bad mental status during the recovery period were harmful to coping with pandemic-related addictive internet use., Scientific Significance: Our findings profiled the general trend of addictive internet use and the vulnerable characteristics of its bad progression across different periods of the first wave of COVID-19 pandemic in China. Our findings provide valuable insights for preventing the long-term adverse effects of negative public events on Internet addiction., (© 2023 The American Academy of Addiction Psychiatry (AAAP).)

This study examined the effect of curcumin on pathological manifestations and clearance of amyloid β peptide (Aβ) in the hippocampus of 8-month-old transgenic APP/PS1 mice with inherent Alzheimer's disease. APP/PS1 mice and the age-matched wild-type controls were subjected to 3 behavioral tests: open field, new object recognition, and Morris water maze. Expression of Aβ, APP, CTF, BACE1, IDE, NEP, and LRP1 proteins in the extracted hippocampal tissue was evaluated by Western blotting. The distribution and the quantity of amyloid plaques and the spread of microglia in the hippocampus were determined by immunofluorescence. The contents of Aβ40 and Aβ42 in the hippocampus were assayed and analyzed on Simoa HD-1 analyzer. The proteins interacting with Aβ in the hippocampus of APP/PS1 mice were detected by co-immunoprecipitation. Curcumin significantly reduced motor hyperactivity in the open-field test, improved short-term recognition memory, spatial learning, and reference memory in APP/PS1 mice. In the hippocampus of APP/PS1 mice, curcumin significantly diminished the elevated Aβ levels and inhibited microglia proliferation. At the same time, curcumin had no effect on Aβ production, extracellular enzymatic hydrolysis, and LRP1-mediated outward transport, but enhanced Aβ clearance by activation of the intracellular ubiquitin-proteasome system and related peripheral mechanisms. Thus, curcumin improves the learning and memory abilities of APP/PS1 mice and reduces the pathological accumulation of Aβ in the brain., (© 2023. Springer Science+Business Media, LLC, part of Springer Nature.)

Fusarium wilt of banana, especially Tropical Race 4 (TR4) is a major factor restricting banana production. Developing a resistant cultivar and inducing plant defenses by elicitor application are currently two of the best options to control this disease. Isotianil is a monocarboxylic acid amide that has been used as a fungicide to control rice blast and could potentially induce systemic acquired resistance in plants. To determine the control effect of elicitor isotianil on TR4 in different resistant cultivars, a greenhouse pot experiment was conducted and its results showed that isotianil could significantly alleviate the symptoms of TR4, provide enhanced disease control on the cultivars 'Baxi' and 'Yunjiao No.1' with control effect 50.14% and 56.14%, respectively. We compared the infection processes in 'Baxi' (susceptible cultivars) and 'Yunjiao No.1' (resistant cultivars) two cultivars inoculated with pathogen TR4. The results showed that TR4 hyphae could rapidly penetrate the cortex into the root vascular bundle for colonization, and the colonization capacity in 'Baxi' was significantly higher than that in 'Yunjiao No.1'. The accumulation of a large number of starch grains was observed in corms cells, and further analysis showed that the starch content in 'Yunjiao No. 1' as resistant cultivar was significantly higher than that in 'Baxi' as susceptible cultivar, and isotianil application could significantly increase the starch content in 'Baxi'. Besides, a mass of tyloses were observed in the roots and corms and these tyloses increased after application with isotianil. Furthermore, the total starch and tyloses contents and the control effect in the corms of 'Yunjiao No.1' was higher than that in the 'Baxi'. Moreover, the expression levels of key genes for plant resistance induction and starch synthesis were analyzed, and the results suggested that these genes were significantly upregulated at different time points after the application of isotianil. These results suggest that there are significant differences between cultivars in response to TR4 invasion and plant reactions with respect to starch accumulation, tyloses formation and the expression of plant resistance induction and starch synthesis related genes. Results also indicate that isotianil application may contribute to disease control by inducing host plant defense against TR4 infection and could be potentially used together with resistant cultivar as integrated approach to manage this destructive disease. Further research under field conditions should be included in the next phases of study., 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 Zhou, He, Tian, Xu, Yang, Liu, Wang, Bai, Li, Li and Zheng.)

Single-atom catalysis efficiently exposes the catalytic sites to reactant molecules while rendering opportunity to investigate the catalysis mechanisms at atomic levels for scientific insights. Here, for the first time, atomically dispersed Co atoms are synthesized as biomimetic "enzymes" to monitor superoxide anions (O 2 •- ), delivering ultraordinary high sensitivity (710.03 μA·μM -1 ·cm -2 ), low detection limit (1.5 nM), and rapid response time (1.2 s), ranking the best among all the reported either bioenzymatic or biomimetic O 2 •- biosensors. The sensor is further successfully employed to real-time monitor O 2 •- released from living cells. Moreover, theoretical calculation and analysis associated with experimental results discover that a mode of end adsorption of the negatively charged O 2 •- on the Co 3+ atom rather than a bridge or/and side adsorption of the two atoms of O 2 •- on two Co 3+ atoms, respectively, plays an important role in the single-atomic catalysis toward O 2 •- oxidation, which not only facilitates faster electron transfer but also offers better selectivity. This work holds great promise for an inexpensive and sensitive atomic biomimetic O 2 •- sensor for bioresearch and clinic diagnosis, while revealing that the adsorption mode plays a critical role in single-atom catalysis for a fundamental insight.

Keloid is a benign skin tumor characterized by its cell hyperproliferative activity, invasion into normal skin, uncontrolled growth, overproduction and deposition of extracellular matrices and high recurrence rate after various therapies. Nintedanib is a receptor tyrosine kinase inhibitor targeting VEGF, PDGF, FGF, and TGF-β receptors with proved efficacy in anti-angiogenesis and in treating various types of cancers. In this study, we investigated the effects of nintedanib on keloid fibroblasts in both in vitro and ex vivo models. Keloid fibroblasts were prepared from 54 keloid scar samples in active stages collected from 49 patients. We found that nintedanib (1-4 μM) dose-dependently suppressed cell proliferation, induced G 0 /G 1 cell cycle arrest, and inhibited migration and invasion of keloid fibroblasts. The drug also significantly inhibited the gene and protein expression of collagen I (COL-1) and III (COL-3), fibronectin (FN), and connective growth factor (CTGF), as well as the gene expression of other pathological factors, such as alpha smooth muscle actin (α-SMA), plasminogen activator inhibitor-1 (PAI-1), FK506-binding protein 10 (FKBP10), and heat shock protein 47 (HSP47) in keloid fibroblasts. Furthermore, nintedanib treatment significantly suppressed the phosphorylation of p38, JNK, ERK, STAT3, and Smad, enhanced endocytosis of various growth factor receptors. Using an ex vivo tissue explant model, we showed that nintedanib significantly suppressed cell proliferation, migration, and collagen production. The drug also significantly disrupted microvessel structure ex vivo. In summary, our results demonstrate that nintedanib is likely to become a potential targeted drug for keloid systemic therapy.

Poly(3,4-ethylenedioxythiophene)/polystyrene sulfonate (PEDOT:PSS) plays an important role in inverted planar perovskite solar cells (IPPSCs) as an efficient hole extraction and transfer layer (HTL). The IPPSCs based on PEDOT:PSS normally display inferior performance with a reduced open-circuit voltage. To address this problem, here sodium citrate-doped PEDOT:PSS is adopted as an effective HTL for improving the performance of IPPSCs. Sodium citrate-doped PEDOT:PSS HTL improves the conversion efficiency of IPPSCs from 15.05% of reference cells to 18.39%. The large increase of the open-circuit voltage ( V OC ) from 1.057 to 1.134 V is the main source for this performance enhancement. With the help of characterization analysis of ultraviolet photoelectron spectroscopy, scanning electron microscopy, electrochemical impedance spectroscopy, etc., the higher work function of the doped PEDOT:PSS film and the uniform crystallinity of the perovskite film on it are disclosed as the reasons for the increased V OC and the consequent performance enhancement.

Background: The clinical outcomes of fat grafting vary and are technique-dependent. Stromal vascular fraction (SVF) gel is a novel, mechanically processed fat product with high concentrations of adipose tissue-derived stem cells and other SVF cells. This study evaluated the volumization and rejuvenation effects of SVF-gel., Objective: This study evaluated the volumization and rejuvenation effects of SVF-gel., Methods: This retrospective, single-center study included 126 patients who underwent SVF-gel grafting and 78 who underwent conventional lipoinjection for various indications from March 2015 to February 2017. Patient satisfaction and secondary surgery rates were evaluated. Samples of transferred SVF-gel were harvested and examined histologically., Results: All patients showed improvements in facial augmentation and contour. Patients in the SVF-gel group experienced mild postoperative swelling and a low secondary surgery rate (10.9%). Assessment of patient-rated satisfaction on a 5-point Likert scale found that 77.3% of patients in the SVF-gel group were satisfied (54.5%) or very satisfied (22.8%) with their outcomes. By comparison, 53.8% of patients who underwent conventional lipoinjection were satisfied (48.7%) or very satisfied (5.1%). Moreover, SVF-gel showed effective antiwrinkle and skin rejuvenation effects. Hematoxylin-eosin staining showed a normal adipose tissue structure in transferred SVF-gel., Conclusion: Stromal vascular fraction gel is effective for both volumization and rejuvenation, and may be superior to conventional lipoinjection for facial recontouring.

Many physical processes such as exciton interfacial dissociation, exciton interfacial recombination, and exciton-electron and exciton-hole interactions coexist at the interface of organic solar cells (OSC). In this study, the direction of free charge generation is defined as the direction from the interface to the side where free charges are left. For a p-n type device, the direction of free electron (hole) generation from exciton dissociation at the donor/accepter (D/A) interface is the same as the subsequent transportation direction under the built-in electric field. However, the direction of free electron (hole) generation from exciton-exciton recombination across the D/A interface is opposite to the direction of free charge transportation. Both free charges generated from exciton interfacial dissociation and recombination are contributed to the photocurrent for a p-n type device. In a device with a heterojunction formed by two n-type materials (here it is defined as an n-n type device), the direction of free electron (hole) generation from exciton recombination across the interface is also the same as the subsequent free charge transportation. At the same time, there are also some free electrons (free holes) generated by exciton interfacial dissociation. The direction of free charge generation from exciton dissociation for this n-n type device is also opposite to the direction of free charge transportation. However, only free charges generated from exciton interfacial recombination are contributed to the photocurrent for an n-n type device. But so far there has been no direct experimental evidence to prove the above theories. In this work, an NPB interfacial layer with a high LUMO was introduced in an n-n type OSC to inhibit the backflow of electrons, which are generated from exciton dissociation at the heterojunction formed by two n-type materials, enhancing the device performance accordingly. This work is conducive to interfacial engineering in an OSC to further improve its performance.

Tissue engineering chamber (TEC) is a technique that could incubate up to 16 folds volume increase of a fat flap. But the mechanism in the silicone chamber was still unknown. The function of exudate in the chamber was noticed recently. We developed a special model called fluid drainage model (FDM) that consisted of a traditional TEC on the back and paired fat flaps without a chamber in the groins. Then we used a silicon tube to dynamically transfer the exudate from dorsal TEC to one of the paired inguinal fat flap and the other inguinal fat flap with a sham tube was set as control. At Week 4, the volume of drainage group reached 8.7 ± 2.3 ml, 576 ± 152% to its original volume whereas the growth ratio of control group was only 130 ± 39%. Similar volume change and histological change were observed within fat flap from TEC model and drainage group. The exudate in the TEC is a heterogeneous cocktail contains cytokines as well as cells. Intriguingly, transferred exudate in the TEC model sustain the ability to incubate large amount of adipose tissue remotely., (Copyright © 2017 John Wiley & Sons, Ltd.)

Background: Hematopoietic stem cells (HSCs) promote fat graft survival by modulating its revascularization. The authors hypothesize that mobilization of HSCs by G-CSF will improve fat graft survival. Hence, we evaluated the effect of different doses of G-CSF on fat grafting., Methods: Male 8-week-old C57 mice received high-dose G-CSF (100 μg/kg), low-dose G-CSF (10 μg/kg), and PBS (control) intraperitoneally for 7 consecutive days right after autologous fat grafting. Grafted fat was harvested at 1, 4, and 12 weeks for examination., Results: The low-dose G-CSF, high-dose G-CSF, and control groups had retention rates of 73.6% ± 3.1%, 51.6% ± 4.4%, and 44.5% ± 4.0%, respectively, at 12 weeks (low-dose G-CSF versus control and low-dose G-CSF versus high-dose G-CSF, both p < 0.05; no significant difference between high-dose G-CSF and control group). Both doses of G-CSF successfully mobilized HSCs into circulation and upregulated the level of blood-derived stem cells in fat grafts, contributing to improved angiogenesis. However, high-dose G-CSF caused a prolonged macrophage infiltration and elevated level of inflammation (IL-6 and TNF-α), which led to severe fibrosis and impaired adipogenesis (downregulated expression of PPAR-γ and CEBP-α)., Conclusions: Low-dose G-CSF treatment successfully improved fat graft survival by mobilizing HSCs and inducing angiogenesis. However, high-dose G-CSF prolonged inflammation and caused severe fibrosis, leading to impaired adipogenesis and poor fat graft survival., (Copyright © 2017. Published by Elsevier Inc.)

Background: Fat grafting experiences a regeneration process from free lipoaspirate to intact adipose tissue. The adipose extracellular matrixes (ECM) provide the structure and biochemical support for surrounding cells; inflammatory cells, like macrophages, regulate the process. Our hypothesis states that transferred fat undergoes ECM remodeling after fat grafting and this process is regulated by macrophage infiltration., Methods: Lipoaspirate was injected subcutaneously into the back of nude mice. The micro-structure of the fat grafts was observed and evaluated using scanning electron microscope (SEM) and collagen I immunohistostaining. The gene transcription level of collagen proteins and the matrix metalloproteinases (MMPs) were assessed by qRT-PCR. Local injection of clodronate-encapsulated liposome was used to evaluate the role of macrophages of fat grafts at different stages in ECM remodeling, depletion of macrophages, at different time points (Week 1 and Week 4)., Results: Results from the SEM analysis showed that liposuction caused severe damage to the ECM structure in freshly aspirated adipose tissue. On Day 1 post-transplantation, the surface of adipocytes was covered with platelets and this secreted fibrin network on the fat grafts. An integral adipose structure was already established with an intact ECM at the end of Week 1. The early depletion of macrophages remarkably hindered ECM reconstruction process by down-regulating the expression of collagen proteins and MMPs. Expression of Collagen I was significantly decreased after depletion of macrophages in both gene and protein levels. Results also showed that the depletion of macrophages at the later stage of fat grafting resulted in less fibrosis and capsule formation., Conclusion: Free fat aspirates undergo a prompt ECM reconstruction process and completed in the first week; this process can be initiated with platelets and mainly modulated by inflammatory cells such as macrophages. It was also observed that prolonged macrophage infiltration contributes to fibrosis and capsule formation in fat grafts., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Purpose: To illustrate the potential for high permittivity materials to be used in decreasing peak local SAR associated with implants when the imaging region is far from the implant., Methods: We performed numerical simulations of a human subject with a pacemaker in a body-sized birdcage coil driven at 128 MHz with and without a thin (5 mm) shell of material of high electric permittivity around the head., Results: For a shell with relative permittivity of 600, the maximum specific energy absorption rate averaged over any 1 g of tissue near the pacemaker was reduced by 73.5% for a given B 1 field strength at the center of the brain., Conclusion: Although further study is required, initial simulations indicate that strategic use of high permittivity materials may broaden the conditions under which patients with certain implants can be imaged safely. Magn Reson Med 78:383-386, 2017. © 2016 International Society for Magnetic Resonance in Medicine., (© 2016 International Society for Magnetic Resonance in Medicine.)

The distribution and function of T lymphocytes in human lung cancer remain limited. In this study, we investigated the properties of human T cell subsets in the blood of non-small cell lung cancer (NSCLC) patients. We found a relatively normal level of CD4+ subsets in the blood of NSCLC patients, but CD8+ effector T cells increased and CD8+ effector memory cells declined compared to the healthy donors. To further analyze their properties, we stimulated the peripheral blood mononuclear cells (PBMCs) of NSCLC patients by mitogens to examine cytokine production. Our data suggest that both CD4+ and CD8+ naïve cells in NSCLC patients significantly reduced IFN-γ and TNF-α production. Additionally, fewer CD8+ effector cells produced IFN-γ and TNF-α in NSCLC patients than in healthy subjects. Moreover, similar results were observed for CD4+ or CD8+ memory cells in NSCLC patients for the production of IFN-γ, TNF-α, and IL-17. Therefore, our results strongly suggest that the function of CD4+ and CD8+ T lymphocytes in NSCLC patients is compromised or dysregulated. The development of vaccines and antitumor immunotherapy may be essential for the treatment of lung cancer patients.

Background: Cell-assisted lipotransfer has been promisingly applied to restore soft-tissue defects in plastic surgery; however, the harvesting of stromal vascular fraction increases morbidity and poses potential safety hazards. The authors investigated whether adding indomethacin, an antiinflammatory proadipogenic drug, to the fat graft at the time of transplantation would enhance the final graft volume compared with cell-assisted lipotransfer., Methods: In vitro, human adipose-derived stem cells were cultured in conditioned growth media supplemented with various doses of indomethacin to investigate adipogenesis and the expression of the adipogenic genes. In vivo, lipoaspirate mixed with stromal vascular fractions or indomethacin was injected into the dorsum of mice. Tissues were harvested at weeks 2, 4, and 12 to evaluate histologic changes., Results: In vitro, polymerase chain reaction analysis revealed that increased up-regulation of adipogenic genes and activation of the peroxisome proliferator-activated receptor-γ pathway. In vivo, the percentage volume of adipocytes in the indomethacin-assisted groups was higher than that in the lipoaspirate-alone (control) group at 12 weeks (p = 0.016), and was equivalent to the volume in the cell-assisted groups (p = 1.000). Indomethacin improved adipose volumes but had no effect on vascularity. A larger number of small adipocytes appeared in the treatment samples than in the controls at 2 weeks (p = 0.044) and 4 weeks (p = 0.021)., Conclusions: Pretreating lipoaspirate with indomethacin enhances the final volume retention of engrafted fat. This result is explained in part by increased adipogenesis and possibly by the inhibition of inflammatory responses.

In plastic and reconstructive field, it would be much beneficial to fabricate an engineered adipose tissue substitute allowing reliable and complete fat tissue regeneration. Tissue engineering chamber (TEC) holds the promise to optimize an adipogenic configuration that is efficacious as well as reproducible. A frequently occurring complication involves the adipose tissue flap encapsulation and, effectively, its shielding, by a thick fibrous membrane, which hinders development into the proliferative stage. The reason for the deposition of the collagen capsule remains unclear. Numerous studies have highlighted that macrophages play a key role in adipogenesis in a TEC model using a silicone chamber enclosing the fat flap with a superficial epigastric pedicle. As a verification of the role of macrophages in capsule formation, we propose the inhibition of transforming growth factor β1 (TGF-β1) synthesis by macrophage populations in the local microenvironment by administrating tranilast into the TEC. We hypothesize that upon reduction of TGF-β1 levels, capsule formation and inhibition of new adipose tissue development will decrease. Furthermore, we propose that a tissue engineering chamber model in which macrophages are closely related to both neo-adipogenesis and capsule formation., (© 2017 International Federation for Cell Biology.)

Aim: The aim of this study was to investigate whether the number of removed lymph nodes was associated with survival of patients with node-negative early cervical cancer and to analyze the prognostic significance of clinical and pathologic features in these patients., Methods: Patients with FIGO stage IA-IIB cervical cancer who underwent radical hysterectomy with lymphadenectomy without receiving preoperative therapy were reviewed retrospectively. Patients were all proved to have lymph-node-negative disease and classified into five groups based on the number of nodes removed. The Kaplan-Meier method and Cox's proportional hazards regression model were used in prognostic analysis., Results: The final dataset included 359 patients: 45 (12.5%) patients had ≤10 nodes removed, 93 (25.9%) had 11-15, 98 (27.3%) had 16-20, 64 (17.8%) had 21-25, and 59 (16.4%) had >25 nodes removed. There was no association between the number of nodes removed and survival of patients with node-negative early cervical cancer (χ 2  = 6.19, P = 0.185). Similarly, subgroup analyses for FIGO stage IB1-IIB also showed that the number of lymph nodes was not significantly related to survival in each stage. Multivariate analyses showed that histology and depth of invasion were independent prognostic factors for survival in these patients., Conclusion: If a standardized lymphadenectomy is performed, the number of lymph nodes removed is not an independent prognostic factor for patients with node-negative early cervical cancer. Our study suggests that there is inconclusive evidence to support survival benefit of complete lymphadenectomy among these patients., (© 2016 Japan Society of Obstetrics and Gynecology.)

In this study, we investigated the effects of remote ischemic preconditioning on post resuscitation cerebral function in a rat model of cardiac arrest and resuscitation. The animals were randomized into six groups: 1) sham operation, 2) lateral ventricle injection and sham operation, 3) cardiac arrest induced by ventricular fibrillation, 4) lateral ventricle injection and cardiac arrest, 5) remote ischemic preconditioning initiated 90min before induction of ventricular fibrillation, and 6) lateral ventricle injection and remote ischemic preconditioning before cardiac arrest. Reagent of Lateral ventricle injection is neuroglobin antisense oligodeoxynucleotides which initiated 24h before sham operation, cardiac arrest or remote ischemic preconditioning. Remote ischemic preconditioning was induced by four cycles of 5min of limb ischemia, followed by 5min of reperfusion. Ventricular fibrillation was induced by current and lasted for 6min. Defibrillation was attempted after 6min of cardiopulmonary resuscitation. The animals were then monitored for 2h and observed for an additionally maximum 70h. Post resuscitation cerebral function was evaluated by neurologic deficit score at 72h after return of spontaneous circulation. Results showed that remote ischemic preconditioning increased neurologic deficit scores. To investigate the neuroprotective effects of remote ischemic preconditioning, we observed neuronal injury at 48 and 72h after return of spontaneous circulation and found that remote ischemic preconditioning significantly decreased the occurrence of neuronal apoptosis and necrosis. To further comprehend mechanism of neuroprotection induced by remote ischemic preconditioning, we found expression of neuroglobin at 24h after return of spontaneous circulation was enhanced. Furthermore, administration of neuroglobin antisense oligodeoxynucleotides before induction of remote ischemic preconditioning showed that the level of neuroglobin was decreased then partly abrogated neuroprotection of remote ischemic preconditioning. These date suggested that neuroglobin involved in neuroprotective effect of remote ischemic preconditioning. In conclusion, remote ischemic preconditioning attenuated post resuscitation cerebral dysfunction and the neuroprotection was mediated partly by high level of neuroglobin in a rat model of cardiac arrest and resuscitation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Purpose: To compare visual quality and quantitative measurements of multislice (MS) breath-hold and 3D respiratory-triggered T1 ρ sequences., Materials and Methods: T1 ρ scans including MS BLOCK T1 ρ, MS HS8 T1 ρ, and 3D BLOCK T1 ρ were performed on 40 healthy volunteers and 17 liver cirrhosis patients on 3.0T magnetic resonance imaging (MRI). The MS breath-hold methods scanned three slices while the 3D method covered 26 slices. Visual scores of image quality, T1 ρ measurements of the three sequences, and T1 ρ values of healthy volunteers and cirrhosis patients were compared using parametric tests., Results: Average visual scores for volunteers and patients of the three sequences were 3.19 and 2.82 for MS BLOCK T1 ρ; 3.71 and 3.59 for MS HS8 T1 ρ; 3.29 and 3.59 for 3D BLOCK T1 ρ, respectively. Difference of visual scores among the three groups was significant (P < 0.001). Differences in T1 ρ values among the three methods were significant (P < 0.001). Difference of T1 ρ between MS BLOCK T1 ρ and 3D BLOCK T1 ρ was not significant (volunteer: P = 0.82, patient: 0.92) while the difference of T1 ρ between MS BLOCK T1 ρ and MS HS8 T1 ρ, MS HS8 T1 ρ and 3D BLOCK T1 ρ was significant in both volunteers and patients (P < 0.01). Differences of T1 ρ between healthy volunteers and patients were all significant in three T1 ρ sequences (P = 0.04, 0.004, 0.03)., Conclusion: Of the tested sequences, the image quality of MS HS8 T1 ρ was best, but 3D BLOCK T1 ρ with respiratory triggering should also be considered as an alternative sequence for clinical diagnosis of liver cirrhosis due to its superior coverage. J. MAGN. RESON. IMAGING 2016;44:906-913., (© 2016 International Society for Magnetic Resonance in Medicine.)

Unlabelled: The aim of this article is to investigate the megakaryopoyesis and thrombopoiesis in preterm infants born to mothers with preeclampsia and the potential effects mediated by soluble fms-like tyrosine kinase 1 (sFlt1) and thrombopoietin (TPO). A perspective case-control study was performed on 26 cord blood of preterm newborns born to mothers with preeclampsia (PE group) and 26 of preterms born to mothers without preeclampsia (control group). Circulating megakaryocyte count and megakaryocyte colony forming units (CFU-MK) were quantified by whole blood infiltration method and plasma clot culture system, respectively. Platelet activation markers, CD62P and CD63, were estimated by flow cytometry. Immunosorbent assays (ELISA) were employed to estimate plasma levels of sFlt1 and TPO of the two groups. When compared to the controls, infants born to mothers with PE had significantly lower peripheral platelet count (PE vs., Controls: 157.9 [44.6] vs. 239.6 [57.5] × 10(9)/l, p < 0.001), circulating MK count (5.8 [1.0] vs. 7.7 [0.9]/ml, p < 0.001) and CFU-MK (14.1 [2.1] vs. 20.1 [2.8]/1 × 10(5) cell, p < 0.001); greater expressions of CD62P (15.5 [2.3] vs. 11.4 [1.9]% platelets, p < 0.001) and CD63 (12.3 [2.4] vs. 9.0 [1.6]% platelets, p < 0.001); increased plasma Flt level (130.1 [8.0] vs. 97.7 [8.7] pg/ml, p < 0.001) and TPO level (129.5 [17.8] vs. 98.9 [11.8] pg/ml, p < 0.001). In PE group, sFlt instead of TPO showed a significantly negative relationship with platelet counts, CFU-MK and circulating MK count, a positive relationship with CD62P, CD63 expressions. In control group, both sFlt and TPO did not show any relationship with these parameters. sFlt played important role in megakaryocytopoesis and platelet homeostasis in preterm infants born to mothers with PE. Its mechanism maybe the effect of impaired megakaryocyte formation and increased platelet activation.

Background: To determine optimal initial age of cardiac iron screening with magnetic resonance imaging (MRI) T2* in patients with thalassemia major (TM)., Methods: We retrospectively reviewed black blood cardiac T2* assessments from 102 TM patients from the ages of 3 to 32 years. Cases of patients under and above 7 years old with detectable cardiac iron overload were analyzed separately. Associations between cardiac T2* and various factors, such as serum ferritin (SF), patient age and hepatic T2*, were assessed using either scatterplots or regression. Images were evaluated by two independent radiologists., Results: With a T2* cut-off value of 20 ms, no patient under 5 years old showed cardiac iron overload. Three of 19 (15.8%) patients under 7 years of age had a cardiac T2* ≤ 20 ms (5.5 to 7 years) but none had ≤10 ms, while 35 of 83 (42.2%) patients above 7 years old had a cardiac T2* ≤ 20 ms (8 to 32 years) and 18 of them ≤10 ms. Cardiac T2* correlated weakly with serum ferritin and liver T2* (r = -0.39 and 0.41, respectively, both P < 0.001), but not with patient age (P > 0.05)., Conclusion: Cardiac iron overload can occur in young TM patients, even as young as 5.5 years old. Assessment of cardiac iron with T2* might need to begin as early as 5 years old if suboptimal chelation therapy is administered., (© 2015 Wiley Periodicals, Inc.)

Introduction: Adipocytes can dedifferentiate into fibroblast-like cells in vitro and thereby acquire proliferation and multipotent capacities to participate in the repair of various organs and tissues. Whether dedifferentiation occurs under physiological or pathological conditions in vivo is unknown., Methods: A tissue expander was placed under the inguinal fat pads of rats and gradually expanded by injection of water. Samples were collected at various time points, and morphological, histological, cytological, ultrastructural, and gene expression analyses were conducted. In a separate experiment, purified green fluorescent protein+ adipocytes were transplanted into C57 mice and collected at various time points. The transplanted adipocytes were assessed by bioluminescence imaging and whole-mount staining., Results: The expanded fat pad was obviously thinner than the untreated fat pad on the opposite side. It was also tougher in texture and with more blood vessels attached. Hematoxylin and eosin staining and transmission electron microscopy indicated there were fewer monolocular adipocytes in the expanded fat pad and the morphology of these cells was altered, most notably their lipid content was discarded. Immunohistochemistry showed that the expanded fat pad contained an increased number of proliferative cells, which may have been derived from adipocytes. Following removal of the tissue expander, many small adipocytes were observed. Bioluminescence imaging suggested that some adipocytes survived when transplanted into an ischemic-hypoxic environment. Whole-mount staining revealed that surviving adipocytes underwent a process similar to adipocyte dedifferentiation in vitro. Monolocular adipocytes became multilocular adipocytes and then fibroblast-like cells., Conclusions: Mature adipocytes may be able to dedifferentiate in vivo, and this may be an adipose tissue self-repair mechanism. The capacity of adipocytes to dedifferentiate into stem cell-like cells may also have a more general role in the regeneration of many tissues, notably in fat grafting.

As a rapid, in-situ analysis method, Field portable X-ray fluorescence spectrometry (FP-XRF) can be widely applied in soil heavy metals analysis field. Whereas, some factors may affect FP-XRF performance and restrict the application. Studies have proved that FP-XRF has poorer performance when the concentration of target element is low, and soil moisture and particle size will affect FP-XRF performance. But few studies have been conducted in depth. This study took an example of Ni, demonstrated the relationship between Ni concentration and FP-XRF performance on accuracy and precision, and gave a critical value. Effects of soil moisture and particle size on accuracy and precision also had been compared. Results show that, FP-XRF performance is related to Ni concentration and the critical value is 400 mg x kg(-1). Relative standard deviation (RSD) and relative uncertainty decrease while the Ni concentration is below 400 mg x kg(-1), hence FP-XRF performance improves with increasing Ni concentration in this range; RSD and relative uncertainty change little while the Ni concentration is above 400 mg x kg(-1), hence FP-XRF performance does not have correlation with Ni concentration any more. For in-situ analysis, the relative uncertainty contributed by soil moisture is 3.77%, and the relative certainty contributed by particle size is 0.56%. Effect of soil moisture is evidently more serious than particle size both on accuracy and precision.

Objectives: The objective of this study is to develop a novel and sensitive method for KRAS codon 12 mutation testing., Design and Methods: We developed a sensitive one-step real-time digestion-and-block TaqMan probe PCR (RTDB-PCR) technique that uses a thermostable endonuclease and a minor groove binder (MGB) blocker to detect KRAS codon 12 mutations. Dilution mimic DNA panels were used to assess the sensitivity of this technique. The RTDB-PCR method was performed and compared with three other methods: PCR sequencing, mutant-enriched PCR sequencing and mutant-enriched PCR-MassArray. A total of 100 formalin-fixed paraffin-embedded (FFPE) metastatic colorectal cancer (mCRC) specimens were also tested by all four methods., Results: The RTDB-PCR was sensitive to as little as 0.01% mutant DNA, significantly higher than other methods. Among the 100 FFPE mCRC specimens examined, 45 tested positive for KRAS codon 12 mutations according to RTDB-PCR, 44 tested positive according to mutant-enriched PCR sequencing and mutant-enriched PCR-MassArray, and only 26 samples tested positive according to PCR sequencing., Conclusions: Compared with mutant-enriched PCR sequencing and mutant-enriched PCR-MassArray, RTDB-PCR is more cost effective, saves time, and is easier to use, making it suitable for the detection of low-level KRAS mutations in the clinic., (Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

Introduction: Glycation products accumulate during aging of slowly renewing tissue, including skin, and are suggested as an important mechanism underlying the skin aging process. Adipose-derived cells are widely used in the clinic to treat ischemic diseases and enhance wound healing. Interestingly, adipose-derived stem cells (ASCs) are also effective in anti-aging therapy, although the mechanism underlying their effects remains unknown. The purpose of the present study was to examine the anti-aging effect of ASCs in a D-galactose-induced aging animal model and to clarify the underlying mechanism., Materials and Methods: Six-week-old nude mice were subcutaneously injected with D-gal daily for 8 weeks. Two weeks after completion of treatment, mice were randomized to receive subcutaneous injections of 106 green fluorescent protein (GFP)-expressing ASCs, aminoguanidine (AG) or phosphate-buffered saline (PBS). Control mice received no treatment. We examined tissue histology and determined the activity of senescence-associated molecular markers such as superoxide dismutase (SOD) and malondialdehyde (MDA)., Results: Transplanted ASCs were detectable for 14 days and their GFP signal disappeared at day 28 after injection. ASCs inhibited advanced glycation end product (AGE) levels in our animal model as well as increased the SOD level and decreased the MDA level, all of which act to reverse the aging phenotype in a similar way to AG, an inhibitor of AGE formation. Furthermore, ASCs released angiogenic factors in vivo such as vascular endothelial growth factor, suggesting a skin trophic effect., Conclusions: These results demonstrate that ASCs may contribute to the regeneration of skin during aging. In addition, the data shows that ASCs provide a functional benefit by glycation suppression, antioxidation, and trophic effects in a mouse model of aging.

The frequent co-existence of arsenic (As) and lead (Pb) necessitates the investigation of clean-up technologies for multi-metal(loid)s. Field survey and hydroponic experiments were conducted to elucidate the co-accumulation of As and Pb in Pteris vittata L. The P. vittata population isolated from a Pb-Zn mine in Yunnan province, China is a potential extractor of As and Pb co-contamination. Hydroponic experiment found that the highest frond As and Pb concentrations in mining population of P. vittata reached 12.2 and 0.99 g kg(-1), respectively. The interaction between As and Pb in P. vittata was further more disclosed. Pb (2 mg L(-1)) improved the frond As concentration by 60 to 150% in mining populations of P. vittata. Micro-X-ray absorption spectroscopy indicated that under the combined exposure of As and Pb, the As content in the rhizoid epidermis increased by about 10-fold, and the As(V) percentage increased in each rhizoid tissue, as compared with that under As exposure alone. The co-absorption of As and Pb on the epidermis and the enhanced transportation of As(V) from epidermis into the rhizoid were suggested to contribute to the increased As accumulation.

Introduction: Adipose tissue transplantation is one of the standard procedures for soft-tissue augmentation, reconstruction, and rejuvenation. However, it is unknown as to how the graft survives after transplantation. We thus seek out to investigate the roles of different cellular components in the survival of graft., Materials & Methods: The ratios of stromal vascular fraction (SVF) cellular components from human adipose tissue were evaluated using flow cytometry. Human liposuction aspirates that were either mixed with marked SVF cells or PBS were transplanted into nude mice. The graft was harvested and stained on days 1,4,7 and 14. The inflammation level of both SVF group and Fat-only group were also evaluated., Results: Flow cytometric analysis showed SVF cells mainly contained blood-derived cells, adipose-derived stromal cells (ASCs), and endothelial cells. Our study revealed that most cells are susceptible to death after transplantation, although CD34+ ASCs can remain viable for 14 days. Notably, we found that ASCs migrated to the peripheral edge of the graft. Moreover, the RT-PCR and the immuno-fluorescence examination revealed that although the SVF did not reduce the number of infiltrating immune cells (macrophages) in the transplant, it does have an immunoregulatory function of up-regulating the expression of CD163 and CD206 and down-regulating that of IL-1β, IL-6., Conclusions: Our study suggests that the survival of adipose tissue after nonvascularized adipose transplantation may be due to the ASCs in SVF cells. Additionally, the immunoregulatory function of SVF cells may be indirectly contributing to the remolding of adipose transplant, which may lead to SVF-enriched adipose transplantation.

In order to disclose soil pollution caused by lead (Pb) smeltery and its human health risks, this study investigated Pb concentrations in farmland soil, hair and blood of residents surrounding a Pb smeltery in Henan Province, and discussed the rationality of estimation of the health protection zone from the Pb smeltery. It was found that the Pb concentrations in blood of children living in both M and Y villages exceeded the international Pb poisoning diagnostic criteria. The highest Pb concentration in blood was 491 microg x L(-1), with the percentages of mild, medium and severe Pb poisoning reaching 52.5%, 42.5% and 5.0%, respectively. Pb concentrations in hair of children living in Y village were in excess of the related standard, with the highest being 156 mg x kg(-1), and the average value 2.9 times of that in hair of adults. In terms of Pb in soil, Pb concentrations in 66.7% of the topsoils (0-20 cm) around the smeltery exceeded Grade II (350 mg x kg(-1), pH > 7.5) of the National Soil Environmental Quality Standard (GB 15618-1995), with the highest reaching up to 1687 mg x kg(-1). The severe soil pollution may have played a role in children's health issues in the villages surrounding the smeltery. Therefore, more attention should be paid to the scientific estimation of health protection zone from Pb smeltery and the remediation of heavy-metal contaminated soil in the surrounding areas.

Optimum perfusion may be the key to the endurance, and hence survival, of autologous adipose tissue transportation. Stromal vascular fraction (SVF) cell therapy can greatly improve the survival of fat grafts by enhancing angiogenesis. However, SVF cells are poorly retained in later stages of SVF-assisted adipose tissue transplant. Therefore, it hardly defines the angiogenic effect through long-term transportation. Adipose tissue suffers from acute hypoxia in the early stage of transportation, leading to the recruitment of macrophages. M2 macrophages enhance angiogenesis in adipose transplantation by acting as an angiogenic signal source, promoting tip cell migration and assisting tip cell fusion. Furthermore, the angiogenic and anti-inflammatory micro-environment in the graft created by M2 macrophages may stimulate the transformation of infiltrating macrophages to M2 macrophages. These M2 macrophages may enhance the long-term retention of graft through angiogenesis. Based on these observations, we postulate that the long-term angiogenic effect of SVF cells may be achieved through the facilitation of the M2 macrophages., (© 2013 International Federation for Cell Biology.)

Background: Accelerated rejection is a type of refractory rejection. Animal models of accelerated rejection are widely employed in research on transplant immunity., Methods: We divided 36 C57BL/6 mice into six groups that underwent heart transplant. To select the ideal number of splenocytes for the presensitization to induce accelerated rejection, were transferred 0.5 × 10(7), 1 × 10(7), 5 × 10(7), or 10 × 10(7) donor splenocytes 7 d before transplantation. We confirmed successful presensitization by increases in donor-reactive antibodies. We performed 12 additional heart transplants in the accelerated rejection group and the control groups for a histological examination, immunohistochemical staining for C3d, and a splenocyte test using flow cytometry., Results: The transfer of 5 × 10(7) donor splenocytes effectively and efficiently induced an accelerated rejection in the BALB/c→C57BL/6 heart transplant, with an allograft survival time that was decreased from 7.4 ± 0.5 d to 3.5 ± 0.8 d compared with the allogenic controls (P < 0.05, log-rank test). An analysis of this model indicated that compared with acute rejection, the number of donor-reactive antibodies was significantly increased, and the proportions of effector memory CD8(+) T cells and plasmacytes in the spleen were significantly increased (7.81% ± 1.2% versus 2.96% ± 1.0%, P = 0.006; 1.27% ± 0.13% versus 0.71% ± 0.22%, P = 0.018, respectively). We found the histological characteristics of both cellular and humoral rejection in the accelerated rejection model., Conclusions: Presensitization via the transfer of donor splenocytes facilitates the establishment of an accelerated rejection model. Our findings with this model indicate that humoral rejection and cellular rejection are coexistent, and that the proportions of effector memory CD8(+) T cells and plasmacytes in the spleen increase significantly during accelerated rejection., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Objective: To study proteins correlated with the mechanical properties of engineered cartilage by screening significantly changed proteins during cartilage formation by comparative proteomic analysis., Methods: Human chondrocyte, cultured and expanded, were seeded onto a polyglycolic acid/polylactic acid (PGA/PLA) scaffolds. After 4 weeks of culture in vitro, the constructs were divided into three groups. There were 6 specimens in each group. For the regular in vitro culture group (A), the constructs were kept in culture at the original condition for an additional 6 weeks. For in vivo groups, the constructs were implanted subcutaneously into nude mice for either 6 weeks (B) or 12 weeks (C). All specimens were harvested for gross observation, average wet weight and volume measurement, histology, immunohistochemistry and biomechanics to evaluate the results. Meanwhile, comparative proteomic analysis was performed for each group, and those proteins involved in extracellular matrix with at least 2 folds up-regulation were chosen for further exploration. The correlations between Young's modulus and the relative content of the selected proteins were analyzed by Pearson correlation coefficient., Results: All these samples in the three groups eventually formed hyaline-like cartilage structure. Specimens in C and B groups were similar with adult articular cartilage in appearance, and had multiple mature lacuna in histology. However, those specimens in A group had loose texture with irregular hypertrophy lacuna. Specimens implanted for 12 weeks in vivo had better wet weight (372.5 +/- 35.4) mg and Young's modulus (8.68 +/- 2.65) MPa than those cultured in vivo for 6 weeks (346 +/- 34.5) mg, (3.25 +/- 1.24) MPa (P < 0.01). In group A, they were (184.4 +/- 12.28) mg and (0.7 +/- 0.23) MPa. This study had detected 44 proteins in ECM by comparative proteomic analysis, then chosing the greatest ratio of 6 up-regulation proteins compared between C and A groups. The correlation results indicated the content of Decorin, Chondroadherin and Fibromodulin were linear correlation with the mechanical properties of engineered cartilage (P < 0.05)., Conclusions: Comparative proteomic analysis could provide large scale information of associated proteins, making it profit for advanced research on the relationship between extracellular matrix and mechanical properties of engineered cartilage by combination with tissue reconstruction techniques.

The present study aimed to investigate the feasibility of isolating adipose-derived stem cells (ADSCs) by selecting cells that express the surface receptor CD105. Surface antigen expression of the unsorted cells was undertaken using FACS analysis. Primary adipose-derived cells were isolated. The second passage cells were incubated with anti-CD105 magnetic beads, and separated using a magnetic separator. Cell growth and colony formation was determined by counting and Giemsa staining, respectively. Cells also underwent histological immunohistochemical, and RT-PCR analyses to determine their chondrogenic, adipogenic and osteogenic potential. Increased cell proliferation and colony formation was observed in CD105-positive (CD105⁺) as compared to the CD105-negative (CD105⁻) cells (P<0.001). Following induction, the expression of type II collagen and the number of calcium deposits and lipid droplets in the CD105⁺ ADCs were markedly higher than in the CD105⁻ ADCs. Furthermore, increased alkaline phosphatase (AKP), leptin and PPARγ2 mRNA expression was detected in the CD105⁺ ADCs (P<0.01). Isolation of CD105⁺ ADSCs by MACS was feasible. Thus, CD105 can be used as a relatively specific marker for the selection of ADSCs. Although the chondrogenic, adipogenic and osteogenic potential of these cells is suggestive of their potential for use in tissue engineering treatments, further in vivo studies are necessary.

Purpose: To assess the time to disease progression (TTP), long-term survival benefit and safety of patients with unresectable hepatocellular carcinoma (HCC) treated with computed tomography (CT)-guided radiofrequency ablation (RFA) with transarterial chemoembolization chemoembolization (TACE)., Methods: This study was approved by the institutional review board. We reviewed the records of patients with intermediate and advanced HCC treated with CT-guided RFA with TACE between January 2000 and December 2009. Median TTP, overall survival (OS) and hepatic function were analyzed with the Kaplan-Meier method and log-rank tests., Results: One hundred and twenty-two patients (112 men and 10 women, mean age 53 years, range 18-86 years) were included in the study. The median follow-up time was 42 months (range 6-89 months), TTP was 6.8 months, the median OS was 31 months, and the 1-, 3-, and 5-year OS were 88.5%, 41.0%, and 10.7%. The results of the univariate analysis revealed that intrahepatic lesion, AJCC stage, and Child-Pugh stage were predictors of OS (P<0.01). In the multivariate analysis, the AJCC stage system showed a statistically significant difference for prognosis. Procedure-related death was 0.21% (1/470) within 1 month, and a statistical difference was found between the TACE and RFA of liver decompensation and Child-Pugh stage (P<0.05)., Conclusions: The survival probabilities of OS increased with CT-guided RFA with TACE, as observed in randomized studies from Europe and Asia. The longest TTP was observed for the intermediate stage HCC. The procedures were well tolerated with acceptable minor and major complications in unresectable HCC patients., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

To study the treatment outcomes of brain-only metastases from small-cell lung cancer (SCLC) at initial diagnose treated by chemotherapy with concurrent brain and thoracic radiotherapy (RT). From Jan 2004 to Jan 2009, 36 treatment-naïve SCLC patients with brain-only metastases in Sun yat-sen University were enrolled. Treatment contained initial EP chemotherapy with concurrent whole-brain radiotherapy (WBRT). EP regimen consisted of etoposide 100 mg/m(2) IV d1-3, cisplatin 80 mg/m(2) IV d1, repeated every 3 weeks. WBRT with total dose of 30 Gy in 10 fractions was started within 1 week from the beginning of chemotherapy followed by thoracic RT including 2 Gy once daily to a total dose of 60 Gy. Treatment responses were evaluated after 3 cycles of chemotherapy. EP regimen was given totally 6 cycles for no tumor progression. Thirty-four patients were evaluable. All of the 20 CNS symptomatic patients experienced symptoms relief. Objective responses in the brain and primary thoracic lesions were observed in 26 (76.5%, 16CR + 10PR) and 29 (85.3%, 23CR + 6PR) patients, respectively. The median survival time (MST) was 19.2 months, and the 1-and 2-year overall survival rates (OS) were 70.6 and 29.4%, respectively, in all patients. Patients with CR response had the longest MST of 21.9 months and 1-and 2-year OS of 93.8 and 43.8%, respectively. Treatment toxicity profiles were acceptable. The treatment strategy of concurrent chemotherapy with brain and thoracic RT might achieve promising survival outcomes comparable to limited-stage SCLC in initially diagnosed SCLC with brain-only metastases.

Acellular cartilage sheets (ACSs) have been used as scaffolds for engineering cartilage with mature chondrocytes. In this study we investigated whether ACSs possess a chondrogenic induction activity that may benefit cartilage engineering with multipotent stem cells. Bone marrow-derived mesenchymal stem cells (BMSCs) isolated from newborn pigs were expanded in vitro and seeded on ACSs that were then stacked layer-by-layer to form BMSC-ACS constructs. Cells seeded on polyglycolic acid/polylactic acid (PGA/PLA) scaffolds served as a control. After 4 weeks of culture with or without additional chondrogenic factors, constructs were subcutaneously implanted into nude mice for another 4 weeks. Cartilage-like tissues were formed after 4 weeks of culture. However, formation of cartilage with a typical lacunar structure was only observed in induced groups. RT-PCR showed that aggrecan, COMP, type II collagen and Sox9 were expressed in all groups except the non-induced BMSC-PGA/PLA group. At 4 weeks post-implantation, cartilage formation was achieved in the induced BMSC-ACS group and partial cartilage formation was achieved in the non-induced BMSC-ACS group, confirmed by safranin O staining, toluidine blue staining and type II collagen immunostaining. In addition, enzyme-linked immunosorbent assay demonstrated the presence of transforming growth factor-β1, insulin-like growth factor-1 and bone morphogenic protein-2 in ACSs. These results indicate that ACSs possess a chondrogenic induction activity that promotes BMSC differentiation., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Induced pluripotent stem (iPS) cells are a potential cell source for regenerative medicine. However, the tumorigenicity of iPS cells is a big concern for clinical application. In addition to the genetic manipulation of the reprogramming process and the greater risk of tumor formation, it is unclear whether iPS cells with normal development potential are still tumorigenic. Here, we investigated 3 mouse iPS cell lines, including one line that is able to generate full-term mice via tetraploid blastocyst complementation. We found that a small number of undifferentiated iPS cells could be steadily isolated and expanded after long-term differentiation of cells in vitro or in vivo. The residual undifferentiated iPS cells could be expanded and redifferentiated, and undifferentiated pluripotent stem cells could again be isolated after further rounds of differentiation, suggesting that residual undifferentiated iPS cells could not be eliminated by extended cell differentiation. The residual undifferentiated cells could form teratomas in vivo, indicating that they are a potential tumorigenic risk during transplantation. These findings prompt us to reconsider the strategies for solving the tumorigenic problem of iPS cells, not only focusing on improving the reprogramming process.

Objective: To explore the feasibility of in vitro chondrogenesis by co-culture of chondrocytes and adipose-derived stromal cells (ADSCs) so as to confirm the hypothesis that chondrocytes can provide chondrogenic microenvironment to induce chondrogenic differentiation of ADSCs., Methods: Human ADSCs and porcine auricular chondrocytes were in vitro expanded respectively and then were mixed at the ratio of 7:3 (ADSCs: chondrocytes). 200 microl mixed cells (5.0 x 10(7)/ml) were seeded onto a polyglycolic acid/polylactic acid (PGA/PLA) scaffold, 8 mm in diameter and 2 mm in thickness, as co-culture group. Chondrocytes and ADSCs with the same cell number were seeded respectively onto the scaffold as positive control group and negative control group. 200 microl chondrocytes (1.5 x 10(7)/ml) were seeded as low concentration chondrocyte group. There were 6 specimens in each group. All specimens were harvested after in vitro culture for 8 weeks in DMEM plus 10% FBS. Gross observation, histology, immunohistochemistry, wet weight measurement and glycosaminoglycan (GAG) quantification were used to evaluate the results. Multiple-sample t-test statistics analysis was done to compare the difference of wet weight and glycosaminoglycan(GAG) content between the groups., Results: Cells in all groups had fine adhesion to the scaffold and could secrete extracellular matrix. In co-culture group and positive control group, cell-scaffold constructs could maintain the original size and shape during in vitro culture. At 8 weeks, cartilage-like tissue formed in gross appearance and histological features, and abundant type II collagen could be detected by immunohistochemistry. Wet weight and glycosaminoglycan(GAG) content of co-culture group were respectively (174 +/- 12) mg and (7.6 +/- 0.4) mg. There were respectively 75% (P < 0.01) and 79% (P<0.01) of those of positive control group. In negative control group, however, constructs shrunk gradually without mature cartilage lacuna in histology. In low concentration chondrocyte group, constructs also shrunk obviously with small amount of cartilage formation at the edge area of the construct, and wet weight was (85 +/- 5) mg, which was 37% (P<0.01) of that of positive control group., Conclusions: Chondrocytes can provide chondrogenic microenvironment to induce chondrogenic differentiation of ADSCs and thus promote the in vitro chondrogenesis of ADSCs.

Human parthenogenetic embryonic stem cells (hpESCs) established from artificially activated oocytes have a wider immune-matching ability because of their homozygosity in the major histocompatibility complex alleles. Whether these cells possess the differentiation capacity similar to regular human embryonic stem cells (hESCs) derived from fertilized eggs is unclear. The aims of this study were to determine whether hpESCs could be differentiated into multipotent mesenchymal stem cell (MSC)-like cells in vitro and then compare these cells with those derived from hESCs. MSC-like cells were obtained from both hpESCs and hESCs, which exhibited similar cell surface marker expression profiles. Further analyses revealed that cells derived from hpESCs possessed stronger osteogenic but weaker adipogenic potentials compared with cells derived from hESCs. This is the first work that demonstrates the differentiation of hpESCs into multipotent MSC-like cells. These hpESCs could be a potential source for cell-based therapies.

Aims: Single-incision laparoscopic procedures are reported to be accessible comprehensively for abdominal surgeries. Herein, we report 1 case of partial hepatectomy in which the single-incision laparoscopic surgery (SILS) technique or the laparoendoscopic single-site surgery was conducted., Case: One 53-year-old woman diagnosed with hepatic multicystis (the largest one had a diameter of 5 cm) underwent single-incision laparoscopic partial hepatectomy on January 1, 2010., Results: The entire procedure was completed in 105 minutes without any complications. The patient went out of bed for mobilization 8 hours after surgery and was discharged on the fifth postoperative day., Conclusion: Single-incision laparoscopic procedures are available for many abdominal surgeries, whereas trials reported to perform partial hepatectomy using SILS are fewer. Therefore, our success in the case of partial hepatectomy by SILS provides another optional approach to liver surgeries.

Objective: To investigate the influence of in vivo or vitro microenvironment on the mechanical properties and histological structure of tissue engineered cartilage, and to provide the appropriate parameters for cartilage construct in vitro., Methods: Human fetal articular chondrocytes were cultured and expanded in vitro, the passage 2 chondrocytes were seeded at the density of 6 x 10(7) cells/cm3 to cylindric dimensional scaffolds made by polyglycolic acid (PGA) and polylactic acid (PLA). These constructs were cultured in vitro for 4 weeks. After 4 weeks, part of samples were implanted subcutaneously into nude mice for 6 and 12 weeks, the others continued to be cultured in vitro. All specimens were harvested after 6 and 12 weeks, and evaluated by gross observation, histology, histochemistry, ultrastructure and mechanical test., Results: All specimens in vivo and vitro eventually formed good shape hyaline cartilage. The constructs in vivo group was white color with smooth surface, and had better mechanical properties than those in vitro, by TEM we can observe the thick and striated collagen fibers in regularly arranged collagen fibril which was similar to adult articular cartilage. The constructs in vitro group was yellow color with coarse surface, the appearance and ultrastructure was similar to fetal articular cartilage. Specimens implanted for 12 weeks in vivo had better compressive modulus(38.28 +/- 3.95) MPa and collagen diameter (41.58 +/- 2.78) nm than those cultured in vitro at the same time (4.12 +/- 0.63) MPa, (15.83 +/- 1.70) nm (P < 0. 01)., Conclusions: The structure and function of human tissue engineered cartilage became mature gradually from vitro to vivo, thick and striated collagen fibrils net similar to adult articular cartilage can be formed in constructs of vivo group,increased collagen cross-linking might be the reason that their mechanical properties been greatly improved.

Acellular cartilage can provide a native extracellular matrix for cartilage engineering. However, it is difficult for cells to migrate into acellular cartilage because of its non-porous structure. The aim of this study is to establish a sandwich model for engineering cartilage with acellular cartilage sheets and chondrocytes. Cartilage from adult pig ear was cut into a circular cylinder with a diameter of approximately 6 mm and freeze-sectioned at thicknesses of 10 μm and 30 μm. The sheets were then decellularized and lyophilized. Chondrocytes isolated from newborn pig ear were expanded for 2 passages. The acellular sheets and chondrocytes were then stacked layer-by-layer, in a sandwich model, and cultured in dishes. After 4 weeks of cultivation, the constructs were then either maintained in culture for another 12 weeks or implanted subcutaneously in nude mouse. Histological analysis showed that cells were completely removed from cartilage sheets after decellularization. By re-seeding cells and stacking 20 layers of sheets together, a cylinder-shaped cell sheet was achieved. Cartilage-like tissues formed after 4 weeks of culture. Histological analyses showed the formation of cartilage with a typical lacunar structure. Cartilage formation was more efficient with 10 μm-thick sheets than with 30 μm sheets. Mature cartilage was achieved after 12 weeks of implantation, which was demonstrated by histology and confirmed by Safranin O, Toluidine blue and anti-type II collagen antibody staining. Furthermore, we achieved cartilage with a designed shape by pre-shaping the sheets prior to implantation. These results indicate that the sandwich model could be a useful model for engineering cartilage in vitro and in vivo., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Objective: To investigate the feasibility of in vitro proliferation of rat Leydig cells by modifying the cell culture system., Methods: Leydig cells were isolated from three-week-old rats by a procedure combining collagenase dispersion, stainless steel mesh infiltration and differential adhesion. The isolated cells were cultured in DMEM/F12 and modified media for stem cell proliferation, and the proliferation of the cultured cells was evaluated by cell counting and MTP test. The expression of 3beta-HSD in the cultured cells was detected by immunohistochemistry and flow cytometry, and testosterone productivity in the isolated Leydig cells with or without hCG stimulation was determined at 2 hours and 4 days after cell isolation., Results: The Leydig cells cultured in the modified media proliferated actively, with a doubling time of (2.26 +/- .31) days, as compared with (16.32 +/- 2.14) days for those cultured in the traditional media (P <0.05). The 3beta-HSD positive rate in the cultured cells was (554.3 +/- 7.1)% after 2 hours and (93.6 +/- 4.6)% after 4 days of culture. All the proliferated cells exhibited testosterone productivity, and their testosterone secretion was significantly upregulated by hCG stimulation (P <0.05)., Conclusion: Leydig cells isolated by differential adhesion proliferate actively in the modified culture media.