BACKGROUND & AIMS: :Magnesium/hydroxyapatite composites were produced by conventional extrusion and their mechanical behavior studied under uniaxial compression at room temperature. The results evidence the capability of the HA for strengthening the Mg material, lowering its microstructural anisotropy and inhibiting deformation twinning. They also reveal that the ECAP processing is effective for improving the grain structure and reducing the crystallographic texture of these composites, giving rise to a significant enhancement of their yield strength and microhardness although the ultimate compressive stress worsens. The analysis of the strain hardening rate of the flow curves demonstrates that the HA addition and the ECAP processing are also effective in inhibiting non-basal dislocation slip.

背景与目标： : 镁/羟基磷灰石复合材料是通过常规挤出生产的，并在室温下在单轴压缩下研究了它们的力学行为。结果证明了HA增强Mg材料，降低其微观结构各向异性并抑制变形孪晶的能力。他们还表明，ECAP工艺可有效改善这些复合材料的晶粒结构并降低其晶体织构，尽管最终的压缩应力会恶化，但会显着提高其屈服强度和显微硬度。对流动曲线的应变硬化率的分析表明，HA的添加和ECAP处理也可以有效抑制非基底位错滑移。

BACKGROUND & AIMS: BACKGROUND:Methylation of CpG sites in genomic DNA plays an important role in gene regulation and especially in gene silencing. We have reported mechanisms of epigenetic regulation for expression of mucins, which are markers of malignancy potential and early detection of human neoplasms. Epigenetic changes in promoter regions appear to be the first step in expression of mucins. Thus, detection of promoter methylation status is important for early diagnosis of cancer, monitoring of tumor behavior, and evaluating the response of tumors to targeted therapy. However, conventional analytical methods for DNA methylation require a large amount of DNA and have low sensitivity. METHODS:Here, we report a modified version of the bisulfite-DGGE (denaturing gradient gel electrophoresis) using a nested PCR approach. We designated this method as methylation specific electrophoresis (MSE). The MSE method is comprised of the following steps: (a) bisulfite treatment of genomic DNA, (b) amplification of the target DNA by a nested PCR approach and (c) applying to DGGE. To examine whether the MSE method is able to analyze DNA methylation of mucin genes in various samples, we apply it to DNA obtained from state cell lines, ethanol-fixed colonic crypts and human pancreatic juices. RESULT:The MSE method greatly decreases the amount of input DNA. The lower detection limit for distinguishing different methylation status is < 0.1% and the detectable minimum amount of DNA is 20 pg, which can be obtained from only a few cells. We also show that MSE can be used for analysis of challenging samples such as human isolated colonic crypts or human pancreatic juices, from which only a small amount of DNA can be extracted. CONCLUSIONS:The MSE method can provide a qualitative information of methylated sequence profile. The MSE method allows sensitive and specific analysis of the DNA methylation pattern of almost any block of multiple CpG sites. The MSE method can be applied to analysis of DNA methylation status in many different clinical samples, and this may facilitate identification of new risk markers.

背景与目标：

BACKGROUND & AIMS: :Current methods for quality control of inactivated influenza vaccines prior to regulatory approval include determining the hemagglutinin (HA) content by single radial immunodiffusion (SRID), verifying neuraminidase (NA) enzymatic activity, and demonstrating that the levels of the contaminant protein ovalbumin are below a set threshold of 1 μg/dose. The SRID assays require the availability of strain-specific reference HA antigens and antibodies, the production of which is a potential rate-limiting step in vaccine development and release, particularly during a pandemic. Immune responses induced by neuraminidase also contribute to protection from infection; however, the amounts of NA antigen in influenza vaccines are currently not quantified or standardized. Here, we report a method for vaccine analysis that yields simultaneous quantification of HA and NA levels much more rapidly than conventional HA quantification techniques, while providing additional valuable information on the total protein content. Enzymatically digested vaccine proteins were analyzed by LC-MS(E), a mass spectrometric technology that allows absolute quantification of analytes, including the HA and NA antigens, other structural influenza proteins and chicken egg proteins associated with the manufacturing process. This method has potential application for increasing the accuracy of reference antigen standards and for validating label claims for HA content in formulated vaccines. It can also be used to monitor NA and chicken egg protein content in order to monitor manufacturing consistency. While this is a useful methodology with potential for broad application, we also discuss herein some of the inherent limitations of this approach and the care and caution that must be taken in its use as a tool for absolute protein quantification. The variations in HA, NA and chicken egg protein concentrations in the vaccines analyzed in this study are indicative of the challenges associated with the current manufacturing and quality control testing procedures.

背景与目标： : 在法规批准之前对灭活流感疫苗进行质量控制的当前方法包括通过单次径向免疫扩散 (SRID) 确定血凝素 (HA) 含量，验证神经氨酸酶 (NA) 的酶活性，并证明污染物的水平卵清蛋白低于设定的阈值1 μ g/剂量。SRID检测需要菌株特异性参考HA抗原和抗体的可用性，其产生是疫苗开发和释放中潜在的限速步骤，尤其是在大流行期间。神经氨酸酶诱导的免疫反应也有助于预防感染; 然而，流感疫苗中NA抗原的量目前尚未量化或标准化。在这里，我们报告了一种疫苗分析方法，该方法比常规HA定量技术更快地同时定量HA和NA水平，同时提供有关总蛋白质含量的其他有价值的信息。酶消化的疫苗蛋白通过lc-ms (E) 进行分析，lc-ms是一种质谱技术，可以对分析物进行绝对定量，包括HA和NA抗原，与制造过程相关的其他结构性流感蛋白和鸡蛋蛋白。该方法在提高参考抗原标准的准确性和验证配方疫苗中HA含量的标签声明方面具有潜在的应用。它也可用于监测NA和鸡蛋蛋白含量，以监测制造一致性。尽管这是一种有用的方法，具有广泛的应用潜力，但我们在此也讨论了该方法的一些固有局限性以及在将其用作绝对蛋白质定量工具时必须采取的谨慎和谨慎。本研究中分析的疫苗中HA，NA和鸡蛋蛋白浓度的变化表明了与当前制造和质量控制测试程序相关的挑战。

BACKGROUND & AIMS: BACKGROUND:Platelets, the smallest human blood cells component, have a key role in the control of haemostasis and thrombosis but they have also been shown to be implicated in a number of different pathological states because of their involvement also in the process of inflammation end its resolution. Their peculiar anucleated morphology render the proteomics an intriguing approach to understand their biology. Given the high impact of platelet in different diseases we have started a systematic investigation of protein repertoire in controlled platelet preparation. MATERIAL AND METHODS:Platelets have been extracted from blood of healthy donors (n=6) collected by venipuncture in Vacutainer. The quality of the preparation was assessed by observation and enumeration in a Bürker chamber with a conventional tissue culture microscope. To characterize human platelets proteome we analysed the pool of purified platelets combining two proteomic approaches: 2-DE separation combined with Mass Spectrometry and nanoscale ultra performances LC-MS(E) shotgun proteomics experiments. RESULTS:The 2D gel analysis leads an average of 1900 protein spots, after the filtering of "noise" and "false positive" spots, over 500 were selected to be eligible for further analysis given their optimal spot quality value. To perform the analysis by ion accounting shotgun proteomic approach, based on nano ultra performance liquid chromatography (nUPLC) coupled to MS(E) processing of continuum LC-MS data, the same pool of samples was subject to liquid phase tryptic digestion and the peptide obtained used for the experiments. All the data obtained were analysed using ProteinLynx GlobalServer v2.3 (PLGS, Waters). Three analytical replicates run were acquire in high/low energy modes and associated to a human protein database returning the identification of 100 distinct genes. Comparative analysis of the Gene Ontology has been performed to evaluate the differential functional representation of the molecular repertoire investigated with these two orthogonal approaches. DISCUSSION:The overall molecular function classification revealed differences between the two proteomic approaches. In particular, we found significant differences in cytoskeletal proteins (19.65% 2-DE versus 45.60 Shotgun) and receptors (0,92% 2-DE versus 6.90% Shotgun).

背景与目标：

BACKGROUND & AIMS: :A novel cleanup technique termed as gas purge-microsyringe extraction (GP-MSE) was evaluated and applied for polycyclic aromatic hydrocarbon (PAH) determination in road dust samples. A total of 68 road dust samples covering almost the entire Shanghai area were analyzed for 16 priority PAHs using gas chromatography-mass spectrometry. The results indicate that the total PAH concentrations over the investigated sites ranged from 1.04μg/g to 134.02μg/g dw with an average of 13.84μg/g. High-molecular-weight compounds (4-6 rings PAHs) were significantly dominant in the total mass of PAHs, and accounted for 77.85% to 93.62%. Diagnostic ratio analysis showed that the road dust PAHs were mainly from the mixture of petroleum and biomass/coal combustions. Principal component analysis in conjunction with multiple linear regression indicated that the two major origins of road dust PAHs were vehicular emissions and biomass/fossil fuel combustions, which contributed 66.7% and 18.8% to the total road dust PAH burden, respectively. The concentration of benzo[a]pyrene equivalent (BaPeq) varied from 0.16μg/g to 24.47μg/g. The six highly carcinogenic PAH species (benz(a)anthracene, benzo(a)pyrene, benzo(b)fluoranthene, benzo(k)fluoranthene, dibenz(a,h)anthracene, and indeno(1,2,3-cd)pyrene) accounted for 98.57% of the total BaPeq concentration. Thus, the toxicity of PAHs in road dust was highly associated with high-molecular-weight compounds.

背景与目标： : 评估了一种称为气体吹扫-微注射器萃取 (gp-mse) 的新型净化技术，并将其用于道路粉尘样品中的多环芳烃 (PAH) 测定。使用气相色谱-质谱法分析了几乎整个上海地区的68个道路粉尘样品中的16个优先PAHs。结果表明，所研究部位的总PAH浓度范围为1.04 μ g/g至134.02 μ g/g dw，平均为13.84 μ g/g。高分子量化合物 (4-6环PAHs) 在PAHs的总质量中占主导地位，占77.85% 至93.62%。诊断比分析表明，道路粉尘PAHs主要来自石油和生物质/煤燃烧的混合物。结合多元线性回归的主成分分析表明，道路粉尘多环芳烃的两个主要来源是车辆排放和生物质/化石燃料燃烧，这分别为道路粉尘多环芳烃总负荷贡献了66.7% 和18.8%。苯并 [a] 芘当量 (BaPeq) 的浓度从0.16 μ g/g变化到24.47 μ g/g。六种高度致癌的PAH物种 (苯并 (a) 蒽，苯并 (a) 芘，苯并 (b) 荧蒽，苯并 (k) 荧蒽，二苯并 (a，h) 蒽和茚并 (1,2，3-cd) 芘占总BaPeq浓度的98.57%。因此，道路粉尘中PAHs的毒性与高分子量化合物高度相关。

BACKGROUND & AIMS: :Male fertility in farm animals is considered as an important economic trait. The phenomenon of spermatogenesis plays a dynamic functional role in determining the viability of sperm and thereby can impact on fertility-driven complications. The process of spermatogenesis is controlled by numerous molecular factors and requires a precisely regulated pattern of gene expression. The role of small noncoding RNAs in altering gene expression has been extensively studied. However, limited information is available apropos their role in yak spermatogenesis. The present study aimed to evaluate the assessment of some significant microRNAs and their expression pattern in the body tissues and sperm of fertile and subfertile yak from Arunachal Pradesh besides identified a novel class of sperm enriched small RNA 'mature-sperm-enriched small RNA' (mse-tsRNA) in Yak spermatozoa. The RNAwas extracted from tissue and sperm using 27 gauge needles and subsequently reverse transcribed into small RNA cDNAs. The PCR positive sperm-predominant miRNAs were validated by quantitative reverse transcriptase PCR (qRT-PCR) for their expression in fertile and subfertile yak. Of the 22 microRNAs, the miRNA19a, miRNA142 and miRNA143 showed higher expression in the subfertile yak, whereas expression of miRNA7d, miRNA23a and miRNA23b were found elevated in the fertile animal. The presence of these small noncoding RNAs in yak sperm and testis indicated the legitimate involvement of their role in yak bull fertility.

背景与目标： : 农场动物的雄性生育能力被认为是重要的经济特征。精子发生现象在决定精子活力方面起着动态功能作用，从而可能影响生育驱动的并发症。精子发生的过程受许多分子因素控制，并且需要精确调节的基因表达模式。小的非编码rna在改变基因表达中的作用已被广泛研究。然而，关于它们在牦牛精子发生中的作用，可获得的信息有限。本研究旨在评估一些重要的microrna及其在阿鲁纳恰尔邦可育和次可育牦牛的身体组织和精子中的表达模式，此外还鉴定了一类新型的精子富集小RNA “成熟精子富集小RNA” (mse-tsrna) 在牦牛精子中。使用27根针头从组织和精子中提取RNA，然后逆转录为小RNA cdna。通过定量逆转录酶PCR (qRT-PCR) 验证了PCR阳性的精子优势mirna在可育和亚育牦牛中的表达。在22个microrna中，miRNA19a，miRNA142和miRNA143在可育的yak中显示出较高的表达，而在可育动物中发现miRNA7d，miRNA23a和miRNA23b的表达升高。牦牛精子和睾丸中这些小的非编码rna的存在表明它们在牦牛生育中的作用是合法的。

BACKGROUND & AIMS: :Complete characterization of microfiltered red-purple pitaya colorant (MRPPC) and its potential applications in foods is described. Using sensorial analysis, products that use carmine or beetroot dye as a food colorant in their formulations were compared. The effect of storage under refrigeration on the microbiological, physicochemical, and chemical changes of MRPPC were evaluated. The results showed that UPLC-ESI-QTOF-MSE was effective for the simultaneous determination of twenty metabolites, putatively identified as carbohydrates, flavonoids, and betalains. The MRPPC was shown to have microbiological and physicochemical stability through twelve weeks of storage, and chemometric analyses efficiently distinguished the metabolic profile in each storage period. Sensory analysis revealed that the MRPPC was useful as a food colorant in yogurt, where it improved color quality without affecting aroma and other characteristics. These results indicate that MRPPC is promising food ingredient as a natural red-purple colorant.

背景与目标： : 描述了微滤红紫火龙果着色剂 (MRPPC) 的完整表征及其在食品中的潜在应用。使用感官分析，比较了在其配方中使用胭脂红或甜菜根染料作为食品着色剂的产品。评估了冷藏储存对MRPPC的微生物，理化和化学变化的影响。结果表明，uplc-esi-QTOF-MSE可有效同时测定20种代谢物，推定为碳水化合物，类黄酮和甜菜碱。MRPPC在储存十二周后显示具有微生物学和理化稳定性，化学计量学分析有效地区分了每个储存期间的代谢特征。感官分析表明，MRPPC可作为酸奶中的食用着色剂，在不影响香气和其他特性的情况下改善了颜色质量。这些结果表明，MRPPC作为天然红紫色着色剂是有希望的食品成分。

BACKGROUND & AIMS: :Let X(0) be an unknown M by N matrix. In matrix recovery, one takes n < MN linear measurements y(1),…,y(n) of X(0), where y(i) = Tr(A(T)iX(0)) and each A(i) is an M by N matrix. A popular approach for matrix recovery is nuclear norm minimization (NNM): solving the convex optimization problem min ||X||*subject to y(i) =Tr(A(T)(i)X) for all 1 ≤ i ≤ n, where || · ||* denotes the nuclear norm, namely, the sum of singular values. Empirical work reveals a phase transition curve, stated in terms of the undersampling fraction δ(n,M,N) = n/(MN), rank fraction ρ=rank(X0)/min {M,N}, and aspect ratio β=M/N. Specifically when the measurement matrices Ai have independent standard Gaussian random entries, a curve δ*(ρ) = δ*(ρ;β) exists such that, if δ > δ*(ρ), NNM typically succeeds for large M,N, whereas if δ < δ*(ρ), it typically fails. An apparently quite different problem is matrix denoising in Gaussian noise, in which an unknown M by N matrix X(0) is to be estimated based on direct noisy measurements Y =X(0) + Z, where the matrix Z has independent and identically distributed Gaussian entries. A popular matrix denoising scheme solves the unconstrained optimization problem min|| Y-X||(2)(F)/2+λ||X||*. When optimally tuned, this scheme achieves the asymptotic minimax mean-squared error M(ρ;β) = lim(M,N → ∞)inf(λ)sup(rank(X) ≤ ρ · M)MSE(X,X(λ)), where M/N → . We report extensive experiments showing that the phase transition δ*(ρ) in the first problem, matrix recovery from Gaussian measurements, coincides with the minimax risk curve M(ρ)=M(ρ;β) in the second problem, matrix denoising in Gaussian noise: δ*(ρ)=M(ρ), for any rank fraction 0 < ρ < 1 (at each common aspect ratio β). Our experiments considered matrices belonging to two constraint classes: real M by N matrices, of various ranks and aspect ratios, and real symmetric positive-semidefinite N by N matrices, of various ranks.

背景与目标： : 设X(0) 为N矩阵的未知M。在矩阵恢复中，对X(0) 进行n δ *(ρ)，则对于大M，N，NNM通常成功，而如果 δ < δ *(ρ)，它通常会失败。一个明显完全不同的问题是高斯噪声中的矩阵去噪，其中将基于直接噪声测量Y = X(0) Z来估计未知的M乘N矩阵X(0)，其中矩阵Z具有独立且相同分布的高斯条目。一种流行的矩阵去噪方案求解无约束优化问题min | | Y-X | |(2)(F)/2 + λ | | X | | *。当进行最佳调整时，该方案实现了渐近极小极大均方误差M(ρ; Β) = lim(M，N → ∞)inf(λ)sup(rank(X) ≤ ρ·M)MSE(X，X(λ))，其中M/N →。我们报告了大量的实验，表明第一个问题中的相变 δ *(ρ)，即从高斯测量中恢复的矩阵，与第二个问题中的最小极大风险曲线M(ρ)= M(ρ; Β) 一致，矩阵去噪高斯噪声: Δ *(ρ)= M(ρ)，对于任何秩分数0 <ρ < 1 (在每个公共纵横比 β 下)。我们的实验考虑了属于两个约束类别的矩阵: 具有不同等级和宽高比的实数M乘N矩阵，以及具有不同等级的实数对称正半定N乘N矩阵。

BACKGROUND & AIMS: :Xueshuantong Lyophilized Powder (XST), consisting of a series of saponins extracted from Panax notoginseng, is widely applied to treat acute cerebral infarction, stroke, and coronary heart disease in China. However, most adverse drug reactions (ADR) in clinic are caused by quality problems of XST. In this study, six batches of certainly abnormal, four batches of possibly abnormal XST, and eight batches of normal XST were obtained from the clinical practice. Their quality fluctuations were identified by ultra-performance liquid chromatography coupled with an electrospray ionization quadrupole time-of-flight mass spectrometry operating in MSE mode (UPLC-QTOF/MSE) and bioassays including antithrombin and proplasmin assay. Fourteen potential components responsible for clinical ADR were identified by UPLC-QTOF/MSE, especially ginsenoside Rg1, Rg3, Rb1 and notoginsenoside R1. In addition, 83.3% (5/6) and 50.0% (3/6) certainly abnormal samples could be identified by UPLC-QTOF/MSE and bioassay, respectively. Interestingly, further integration of the two methods could entirely identify all the certainly abnormal samples and inferred that all the possibly abnormal samples were closely related to their quality fluctuation. It indicates that it is advisable to combine UPLC-QTOF/MSE and bioassay for identifying quality fluctuation of XST, and thus reduce its ADR in clinic.

背景与目标： : 血栓通冻干粉 (XST) 由从三七中提取的一系列皂苷组成，在中国广泛用于治疗急性脑梗死，中风和冠心病。然而，临床上大多数药品不良反应 (ADR) 是由XST的质量问题引起的。在这项研究中，从临床实践中获得了六批肯定异常，四批可能异常的XST和八批正常的XST。通过超高效液相色谱法与以MSE模式 (uplc-qtof/MSE) 运行的电喷雾电离四极杆飞行时间质谱法以及包括抗凝血酶和原纤溶酶测定在内的生物测定法，可以鉴定其质量波动。Uplc-qtof/MSE鉴定出14种可能导致临床ADR的成分，尤其是人参皂苷Rg1，Rg3，Rb1和三七皂苷r1。此外，83.3% (5/6) 和50.0% (3/6) 当然异常样品可以分别通过uplc-qtof/MSE和生物测定来鉴定。有趣的是，两种方法的进一步整合可以完全识别所有肯定的异常样本，并推断所有可能的异常样本与其质量波动密切相关。这表明建议将uplc-qtof/MSE与生物测定相结合以识别XST的质量波动，从而减少其在临床上的不良反应。

BACKGROUND & AIMS: :Characterization of phospholipids (PLs) and phytosterols were determined in Schisandra chinensis (S. chinensis) oil by UPLC-Q/TOF-MSE. The determination process was based on feature fragment information of components generated by the MSE detector. A total of 49 and 39 PLs were identified in S. chinensis oil under negative and positive ion mode, respectively. The classes of PLs included phosphatidic acids (PAs), phosphatidylethanolamines (PEs), phosphatidylglycerols (PGs), phosphatidylinositols (PIs), phosphatidylserines (PSs) and phosphatidylcholines (PCs). The most diverse species of PLs detected were PIs and PCs, accounting for 12 and 14, respectively. The analysis of quantification indicated that PEs and PCs were the most abundant constituents in S. chinensis oil, accounting for 122.85 ± 3.82 and 85.61 ± 2.12 μg/g, respectively. Besides, thirteen kinds of phytosterols were tentatively identified in S. chinensis oil under positive ion mode, among which, conicasterol C was the most abundant component, accounting for 22.02 ± 0.98 μg/g. Brassicasterol, campesterol, secosterol-B and herbasterol were also determined in S. chinensis oil. These results might be meaningful in the quality assessment and function evaluation of S. chinensis oil.

背景与目标： : 用uplc-q/TOF-MSE测定五味子油中磷脂 (PLs) 和植物甾醇的表征。确定过程基于MSE检测器生成的组件的特征片段信息。在负离子和正离子模式下，分别在中华沙门氏菌油中鉴定出49个和39个pls。PLs的类别包括磷脂酸 (PAs)，磷脂酰乙醇胺 (PEs)，磷脂酰甘油 (PGs)，磷脂酰肌醇 (PIs)，磷脂酰丝氨酸 (PSs) 和磷脂酰胆碱 (PCs)。检测到的PLs种类最多的是PIs和PCs，分别占12和14。定量分析表明，PEs和PCs是香豆油中含量最高的成分，分别占122.85   ±   3.82和85.61   ±   2.12  μ g/g。此外，在正离子模式下，初步鉴定了十三种植物甾醇，其中以conicasterol C含量最高，占22.02   ±   0.98  μ g/g。还测定了中华沙门氏菌油中的油菜甾醇，菜油甾醇，甾醇-B和草本甾醇。这些结果可能对香豆油的质量评估和功能评估具有重要意义。

BACKGROUND & AIMS: :Xenotransplantation using pig organs has achieved survival times up to 195 days in pig orthotopic heart transplantation into baboons. Here we demonstrate that in addition to an improved immunosuppressive regimen, non-ischaemic preservation with continuous perfusion and control of post-transplantation growth of the transplant, prevention of transmission of the porcine cytomegalovirus (PCMV) plays an important role in achieving long survival times. For the first time we demonstrate that PCMV transmission in orthotopic pig heart xenotransplantation was associated with a reduced survival time of the transplant and increased levels of IL-6 and TNFα were found in the transplanted baboon. Furthermore, high levels of tPA-PAI-1 complexes were found, suggesting a complete loss of the pro-fibrinolytic properties of the endothelial cells. These data show that PCMV has an important impact on transplant survival and call for elimination of PCMV from donor pigs.

背景与目标： : 使用猪器官的异种移植在猪原位心脏移植到狒狒中获得了长达195天的存活时间。在这里，我们证明，除了改进的免疫抑制方案，持续灌注的非缺血保存和移植后生长的控制外，预防猪巨细胞病毒 (PCMV) 的传播在实现长存活时间中起着重要作用。我们首次证明了原位猪心脏异种移植中PCMV的传播与移植的存活时间减少有关，并且在移植的狒狒中发现了IL-6和tnf α 水平的升高。此外，发现了高水平的tPA-PAI-1复合物，表明内皮细胞的促纤溶特性完全丧失。这些数据表明，PCMV对移植存活具有重要影响，并呼吁从供体猪中消除PCMV。

BACKGROUND & AIMS: BACKGROUND AND PURPOSE:Leptin is important in the regulation of fat mass and body weight. Adipose tissue not only secretes leptin but also serves as a site of action for leptin. This study was designed to examine the relationships among tissue expression of leptin receptors, serum leptin, and body mass index. METHODS:Omental adipose tissue and fasting blood samples were obtained from 57 nondiabetic women who underwent surgery for either myoma of the uterus or ovarian cyst. Tissue RNA was extracted using Trizol reagent and serum leptin concentrations were determined with commercial kits. The leptin receptor isoforms in tissues were quantified using real-time Taqman technology. RESULTS:Three leptin receptor isoforms, Ob-Rb, HuB219.1, and HuB219.3, were found in human omental adipose tissue. The amounts of HuB219.1 and HuB219.3 mRNA relative to that of Ob-Rb were 1314.2 and 16.7, respectively. Higher body mass index was significantly correlated with an increase in serum leptin concentration and a decrease in leptin receptor HuB219.1 isoform in omental fat, even after adjustment for age and menopausal status. There was no direct association between serum leptin concentration and tissue HuB219.1 mRNA level. CONCLUSIONS:HuB219.1 is the major isoform of leptin receptor expressed in human omental adipose tissue. Our findings suggest that the shorter leptin receptor isoforms in human omental adipose tissue might play an important role in body weight control. Further studies on the inter-relationship between leptin concentrations and multiple leptin receptor isoforms are needed to elucidate the exact mechanism of obesity.

背景与目标：

BACKGROUND & AIMS: :Carbonic anhydrase III (CAIII) is a metabolic enzyme and a regulator for intracellular pH. CAIII has been reported with high level expression in slow twitch skeletal muscles. Here we demonstrate that CAIII is expressed in multiple slow and fast twitch muscles of adult mouse independent of the expression of myosin isoforms. Expressing similar fast type of myofilament proteins, CAIII-positive tibial anterior (TA) muscle exhibits higher tolerance to fatigue than that of CAIII-negative fast twitch extensor digitorum longus (EDL) muscle in in situ contractility studies. We further studied the muscles of CAIII knockout (Car3-KO) mice. The loss of CAIII in soleus and TA muscles in Car3-KO mice did not change muscle mass, sarcomere protein isoform contents, and the baseline twitch and tetanic contractility as compared with age-matched wild type (WT) controls. On the other hand, Car3-KO TA muscle showed faster force reduction at the beginning but higher resistance at the end during a fatigue test, followed by slower post fatigue recovery than that of WT TA muscle. Superfused Car3-KO soleus muscle also had faster total force reduction during fatigue test than that of WT soleus. However, it showed a less elevation of resting tension followed by a better post fatigue recovery under acidotic stress. CAIII was detected in neonatal TA and EDL muscle, downregulated during development, and then re-expressed in adult TA but not EDL muscles. The expression of CAIII in Tnnt1-KO myopathy mouse soleus muscle that has diminished slow fiber contents due to the loss of slow troponin T remained high. Car3-KO EDL, TA, and soleus muscles showed no change in the expression of mitochondria biomarker proteins. The data suggest a fiber type independent expression of CAIII with a role in the regulation of intracellular pH in skeletal muscle and may be explored as a target for improving fatigue resistance and for the treatment of TNNT1 myopathies.

背景与目标： 碳酸酐酶III (caii) 是一种代谢酶，是细胞内pH的调节剂。据报道，caii在缓慢抽搐的骨骼肌中具有高水平表达。在这里，我们证明caii在成年小鼠的多个慢和快抽动肌肉中表达，而与肌球蛋白同工型的表达无关。在原位收缩力研究中，caii阳性胫骨前 (TA) 肌表达相似的快速类型的肌丝蛋白，对乏力的耐受性高于caii阴性的快速抽搐指长伸肌 (EDL)。我们进一步研究了caii基因敲除 (Car3-KO) 小鼠的肌肉。与年龄匹配的野生型 (WT) 对照相比，Car3-KO小鼠比目鱼肌和TA肌肉中caii的丧失没有改变肌肉质量，肌节蛋白同工型含量以及基线抽搐和强直收缩力。另一方面，在乏力测试期间，Car3-KO TA肌肉在开始时显示出更快的力降低，但在结束时显示出更高的阻力，随后比WT TA肌肉的疲劳后恢复更慢。与比目鱼肌相比，Car3-KO比目鱼肌在乏力测试期间的总力降低也更快。然而，它显示出较低的静息张力升高，然后在酸中毒压力下乏力后恢复。Caii在新生儿TA和EDL肌肉中检测到，在发育过程中下调，然后在成年TA中重新表达，但在EDL肌肉中不表达。由于肌钙蛋白T的丢失，Tnnt1-KO肌病小鼠比目鱼肌中caii的表达降低了慢纤维含量，但仍很高。Car3-KO EDL，TA和比目鱼肌显示线粒体生物标志物蛋白的表达没有变化。数据表明caii的纤维类型独立表达在骨骼肌细胞内pH的调节中起作用，可以作为改善乏力性和治疗TNNT1肌病的靶标。

BACKGROUND & AIMS: :Chronic cutaneous disease of mice caused by the protozoan parasite Leishmania mexicana requires interleukin-10 (IL-10) and FcγRIII (an activating IgG receptor). Macrophages readily secrete IL-10 in response to IgG-coated amastigotes, making macrophages a prime candidate as the critical source of IL-10. However, indirect evidence suggested that macrophage IL-10 is not essential for chronic disease. I now show directly that mice lacking IL-10 from macrophages and granulocytes still have chronic disease, like wild-type C57BL/6 mice. However, T cell-derived IL-10 is required for chronic disease. CD4-cre IL-10flox/flox mice lack IL-10 from T cells (both CD4+ and CD8+) and heal their L. mexicana lesions, with parasite control. I had previously shown that depletion of CD25+ T cells had no effect on chronic disease, and thus, T cells other than CD25+ regulatory T (Treg) cells should be the important source of IL-10. Given that conventional T cells do not express FcγRs, there is likely to be an indirect pathway by which FcγRIII on some other cell engaged by IgG1-amastigote immune complexes induces IL-10 from T cells. Further work is needed to delineate these pathways.

背景与目标： : 由原生动物寄生虫利什曼原虫引起的小鼠慢性皮肤病需要interleukin-10 (IL-10) 和fc γ riii (激活的IgG受体)。巨噬细胞容易分泌IL-10，以响应IgG包被的amastiges，使巨噬细胞成为作为关键IL-10来源的主要候选者。然而，间接证据表明巨噬细胞IL-10对于慢性疾病不是必需的。我现在直接显示缺乏巨噬细胞和粒细胞IL-10的小鼠仍然有慢性疾病，像野生型C57BL/6小鼠。然而，慢性疾病需要T细胞衍生的IL-10。CD4-cre IL-10flox/flox小鼠缺乏来自T细胞 (CD4和CD8) 的IL-10，并通过寄生虫控制治愈其墨西哥假单胞菌病变。我先前已经证明CD25 T细胞的耗竭对慢性疾病没有影响，因此，除CD25调节性T (Treg) 细胞以外的T细胞应该是IL-10的重要来源。鉴于常规T细胞不表达fc γ rs，可能存在一种间接途径，通过该途径，IgG1-amastigote免疫复合物参与的某些其他细胞上的fc γ riii诱导T细胞IL-10。需要进一步的工作来描述这些途径。

BACKGROUND & AIMS: :Warfarin induced skin necrosis occurs in 0.01-0.1% of warfarin treated patients. The usual presentation is that of painful lesions developing in obese women after the initiation of warfarin treatment. The lesions usually evolve into full thickness skin necrosis within a few days. Although the exact mechanism is not totally clear, low levels of Protein C or S, either functional or inherited, are associated with many of the cases. We report the case of a 17-year-old patient treated with warfarin because of iliofemoral deep vein thrombosis post abortion. The patient developed several huge haemorrhagic blisters on the affected leg. The condition rapidly developed into full thickness skin and fat necrosis. The necrotic lesions were excised and eventually covered with skin graft. The combination of the patient tendency towards hyper-coagulation and the local factors is discussed.

背景与目标： 华法林诱导的皮肤坏死发生在华法林治疗患者的0.01 0.1%。通常的表现是开始使用华法林治疗后肥胖女性出现的疼痛性病变。病变通常在几天内演变为全层皮肤坏死。尽管确切的机制尚不完全清楚，但许多情况下，功能性或遗传性的蛋白质C或S水平较低。我们报告了一例17岁的患者，该患者因流产后的股静脉深静脉血栓形成而接受华法林治疗。患者在患肢上出现了几个巨大的出血性水泡。病情迅速发展为全层皮肤和脂肪坏死。切除坏死性病变，并最终覆盖皮肤移植物。讨论了患者的高凝倾向和局部因素的结合。