Association between ADIPOQ gene polymor-phisms and the risk of new-onset diabetes mel-litus after liver transplantation

BACKGROUND:New-onset diabetes after transplantation (NODAT) has become one of the major factors that affect the overall survival and long-term life quality in liver transplan-tation (LT) recipients. Previous studies found that the serum adiponectin concentration of diabetic patients is significantly lower than that of healthy subjects. Adiponectin regulates the blood glucose level by increasing body sensitivityto insulin through various mechanisms. In this study, we aimed to in-vestigate the impact of diabetes related gene polymorphisms on the development of NODAT in liver recipients. METHODS: A total of 256 LT patients in a single-center were selected retrospectively for the study. Genomic DNA was extracted from explanted liver tissues, and tested for twelve diabetes mellitus associated single nucleotide polymorphisms by Sequenom MassARRAY. Modified clinical models in pre-dicting NODAT were established and evaluated.RESULTS: The GG genotype of ADIPOQ rs1501299 gene polymorphism was significantly more frequent in NODAT than non-NODAT LT patients (56% vs 39%,P=0.014). Dom-inant model (GG vs GT+TT,P=0.030) and recessive model (GT+GG vs TT,P=0.005) also confirmed the genotype distri-bution difference between NODAT and non-NODAT groups. Age (OR=1.048,P=0.004), BMI (OR=1.107,P=0.041), and blood tacrolimus level at 1-month LT (OR=1.170,P=0.003) were clinical independent risk factorsof NODAT. Furthermore, rs1501299 could improve the ability of clinical model in predict-ing NODAT (AUROC=0.743,P<0.001). CONCLUSION: ADIPOQ rs1501299 gene polymorphism is associated with an increased risk of NODAT, which should be added to the clinical models in predicting the occurrence of NODAT in LT recipients.     

MCS＋血细胞分离机在治疗性红细胞采集术中的应用

目的探讨MCS＋血细胞分离机应用于治疗性红细胞采集（TE）时的操作要点和临床观察指标。方法采用治疗性自体红细胞采集（TAE）程序，选择随机等量补充补偿液，不加添加剂模式。比较采集前后的红细胞（RBC）、红细胞比积（HCT）、血红蛋白（HGB）、B超等结果，观察治疗效果；分析比较机器显示的预计采后HCT值与实际采后的外周血HCT的差别。结果6例病人采集后外周血血常规（血Rt）示RBC、HCT、HGB均明显下降，与采集前比较差异有统计学意义（P〈0．01），原有脾肿大的患者B超均明显缩小；机器显示的预计采后HCT均比实际采后外周血HCT低，差异有统计学意义（P〈0．01）。结论应用MCS＋血细胞分离机进行他是安全有效的。     

造影血管边缘检测方法研究

本文讨论了一种新的血管造影图像边缘检测方法。这一方法是首先调整正规化滤波器的正规化参数用以平滑血管造影图像，去除低于这参数尺度的细节噪声，再利用小波变换提取血管的边缘。我们的结果表明，在相当大的正规化参数范围内，有效地去除不需要的细节噪声的同时，还能很好的保证提取的血管边缘的精度。与单独使用小波变换或递推正规化滤波器相比，该方法的优点是既弥补了小波变换去除相关噪声差的弱点，又改善了正规化滤波器局部分辨率差的缺点。     

Development and characterization of an SV40 immortalized porcine ameloblast-like cell line

Enamel is secreted as a protein matrix by the ameloblasts. These same cells then control the maturation of the enamel matrix, secreting proteinases that hydrolyze proteins as mineralization progresses, until mature enamel containing less than 1% protein by weight remains. Further understanding of the factors that control ameloblast function and differentiation requires an in vitro cell culture system. In this study, we report immortalization of enamel organ epithelial cells and the selection of a cell line with characteristics of ameloblasts. Porcine enamel organ cells were dissected from unerupted porcine molars, cultured in serum-free medium, and passaged twice. These cells were transfected with an origin-of-replication defective SV40 plasmid by calcium phosphate precipitation, and a cell line with mRNA expression characteristic of ameloblasts was cloned. This cell line (PABSo-E) expressed mRNA for amelogenin, matrix metalloproteinase-20 (enamelysin), and enamel matrix serine proteinase 1 (EMSP1), but not ameloblastin. PABSo-E cells have been passaged more than 55 times, while continuing to maintain characteristics of ameloblasts. These cells will be useful for future studies of ameloblast function.     

年轻子宫内膜癌患者保守治疗新进展

子宫内膜癌在全球范围内呈现出年轻化的发病趋势,严重危害育龄期妇女健康。保守治疗越来越多地用于希望保留生育功能的年轻子宫内膜癌患者,但仍面临着肿瘤复发及耐药等问题。近年来,为避免以上问题及寻找新的保守治疗方式,开展了一系列的研究。该文将有关年轻子宫内膜癌保守治疗的最新研究进行综述,旨在为年轻子宫内膜癌的临床治疗提供参考。     

磁共振检查在妊娠期的应用

妊娠期磁共振(magnetic resonance,MR)检查可以作为超声的有益补充,已广泛应用于非产科、产科母体及胎儿疾病。妊娠期MR检查相对安全,在孕妇或胎儿的获益高于潜在风险时,可以进行MR平扫检查。妊娠期含钆对比剂的使用会增加胎死宫内和新生儿死亡等不良事件的发生风险,因此不推荐妊娠期常规使用含钆对比剂。MR检查的指征和扫描方案在妊娠期可根据需要进行一定调整。MR检查软组织分辨率高、没有电离辐射,在妊娠期可以替代部分辐射性检查用于妊娠合并肿瘤患者。妊娠期母体合并急腹症、瘢痕妊娠和胎盘植入等,以及胎儿疾病,MR检查均可以作为超声的补充,有助于诊断和妊娠期管理。     

Acquisition of the arginine deiminase system benefits epiparasitic Saccharibacteria and their host bacteria in a mammalian niche environment

Saccharibacteria are a group of widespread and genetically diverse ultrasmall bacteria with highly reduced genomes that belong to the Candidate Phyla Radiation. Comparative genomic analyses suggest convergent evolution of key functions enabling the adaptation of environmental Saccharibacteria to mammalian microbiomes. Currently, our understanding of this environment-to-mammal niche transition within Saccharibacteria and their obligate episymbiotic association with host bacteria is limited. Here, we identified a complete arginine deiminase system (ADS), found in further genome streamlined mammal-associated Saccharibacteria but missing in their environmental counterparts, suggesting acquisition during environment-to-mammal niche transition. Using TM7x, the first cultured Saccharibacteria strain from the human oral microbiome and its host bacterium Actinomyces odontolyticus, we experimentally tested the function and impact of the ADS. We demonstrated that by catabolizing arginine and generating adenosine triphosphate, the ADS allows metabolically restrained TM7x to maintain higher viability and infectivity when disassociated from the host bacterium. Furthermore, the ADS protects TM7x and its host bacterium from acid stress, a condition frequently encountered within the human oral cavity due to bacterial metabolism of dietary carbohydrates. Intriguingly, with a restricted host range, TM7x forms obligate associations with Actinomyces spp. lacking the ADS but not those carrying the ADS, suggesting the acquired ADS may also contribute to partner selection for cooperative episymbiosis within a mammalian microbiome. These data present experimental characterization of a mutualistic interaction between TM7x and their host bacteria, and illustrate the benefits of acquiring a novel pathway in the transition of Saccharibacteria to mammalian microbiomes.

The recent discovery of the Candidate Phyla Radiation (CPR), a large monophyletic radiation of phyla and superphyla that includes the group currently referred to as Patescibacteria (1) and accounts for over 26% of microbial diversity, has greatly expanded the bacterial tree of life (2–4). Among the CPR (>73 phyla), Saccharibacteria (formerly known as TM7) represents one of the only three phyla, the other two being uncultivated Candidatus Absconditabacteria and Candidatus Gracilibacteria, whose members are detected in mammalian microbiomes in addition to diverse environmental niches (5, 6). Analyses suggest multiple acquisition events (transfer of bacteria from the environment to mammals that results in stable incorporation into the mammalian microbiome) in the past led to the current wide diversity, with convergent evolution of key functions allowing Saccharibacteria from the environment with already extremely reduced genomes to adapt to mammals (7). There are six main phylogenetic groups (G1 to G6) identified within the Saccharibacteria, with only the G1 group containing both mammalian and environmental representatives. Remarkably low variation and high synteny in genomes between human-associated and groundwater Saccharibacteria have been observed for the G1 lineage, suggesting their more recent acquisition and adaptation from environmental to mammalian hosts (7). A subsequent study further suggested that other mammal-associated CPR species also originated in the environment, with the oral cavity being their most abundant habitat within animal hosts (6). Furthermore, recent studies revealed the habitat-specific difference in gene content among the CPR (5–8). Some of the highly enriched functions among mammal-associated CPR species have been implicated in either enabling use of eukaryotic host-specific resources or conferring tolerance to host-specific stressors (6). Despite these findings, the paucity of cultivated representatives has impeded the experimental evaluation of the key functions acquired by CPR species during their transition from environments to mammals.The first cultivated representative of Saccharibacteria and the CPR group, Nanosynbacter lyticus strain TM7x, was coisolated with its host bacterium Actinomyces odontolyticus strain XH001 (referred to as XH001 in this study) from the human oral cavity (9). TM7x has an ultrasmall cell size (200 to 300 nm), and a reduced genome of only 705 genes with limited de novo biosynthetic capabilities (9, 10). Initial characterization revealed a unique lifestyle of TM7x as an obligate epiparasite grown on the surface of its host bacterium XH001, gaining essential growth requirements at the expense of XH001 under typical laboratory conditions (10, 11). Furthermore, TM7x cells can dissociate from their host bacteria for horizonal transmission when compatible species/strains are present (11). Accumulating evidence indicates that CPR organisms have shared characteristics such as ultrasmall cell sizes, reduced genomes with minimal biosynthetic capabilities, and physical association with prokaryotic host cells, suggesting that symbiosis could be a common lifestyle shared by these diverse while phylogenetically related bacterial groups (4, 8). Cultivation of additional human oral Saccharibacteria (12) has further confirmed the generality of an obligate symbiotic lifestyle and allows the possibility of investigating the specific functions enabling their adaptation to animal-associated habitats and, more broadly, their relationship with their host bacteria.In this study, using TM7x and its host bacterium XH001 as a model system, we experimentally investigated the functions likely acquired during the transition of Saccharibacteria from the environment and adaptation to a mammalian niche. Despite having highly reduced genomes, a complete arginine deiminase system (ADS), an arginine catabolism pathway encoded by a four-gene locus, is found in the majority of mammal/human-associated, but not environmental, Saccharibacteria strains. We show that TM7x encodes an intact and functional ADS, which allows TM7x cells to catabolize arginine, an amino acid readily available within the human oral cavity (13), and generate intermediate and final catabolites including adenosine triphosphate (ATP) and ammonia. Importantly, the ADS maintains the membrane integrity and infectivity of TM7x cells when they are dissociated from their host bacteria, thus facilitating their transmission to other host bacteria and ensuring their propagation. Furthermore, the TM7x-encoded ADS confers a shared benefit to both TM7x and its host bacterium by protecting both partners from acid stress. Intriguingly, TM7x can specifically form a symbiotic relationship with Actinomyces spp. lacking the ADS (including XH001) but not those carrying an ADS pathway, suggesting the ADS in TM7x may be important for TM7x–host bacteria coevolution. Our data presented experimental evidence demonstrating the benefit conferred by an acquired function (e.g., the ADS) in Saccharibacteria species that facilitates their adaptation from the environment to a mammalian niche.     

Acute kidney injury defined by cystatin C may be superior for predicting the outcomes of liver cirrhosis with acute gastrointestinal bleeding

Background & AimsAcute kidney injury (AKI) is conventionally evaluated by a dynamic change of serum creatinine (Scr). Cystatin C (CysC) seems to be a more accurate biomarker for assessing kidney function. This retrospective multicenter study aims to evaluate whether AKI re-defined by CysC can predict the in-hospital outcomes of patients with liver cirrhosis and acute gastrointestinal bleeding.MethodsOverall, 677 cirrhotic patients with acute gastrointestinal bleeding, in whom both Scr and CysC levels were detected at admissions, were screened. eGFR_Scr, eGFR_CysC, and eGFR_Scr-CysC were calculated. MELD-Na score and AKI were re-evaluated by CysC instead of Scr. Odds ratios (ORs) were calculated in the logistic regression analyses. The receiver operating characteristic (ROC) curve analyses were performed.ResultsUnivariate logistic regression analyses demonstrated that baseline Scr and CysC levels, eGFR_Scr, eGFR_CysC, eGFR_Scr-CysC, original MELD-Na score defined by Scr, MELD-Na score re-defined by CysC, and AKI re-defined by CysC, but not conventional AKI defined by Scr, were significantly associated with in-hospital death. ROC analyses showed that baseline CysC level, eGFR_Scr, eGFR_CysC, eGFR_Scr-CysC, original MELD-Na score defined by Scr, and MELD-Na score re-defined by CysC, but not baseline Scr level, could significantly predict the risk of in-hospital death.ConclusionsAKI re-defined by CysC may be superior for predicting the in-hospital mortality of cirrhotic patients with acute gastrointestinal bleeding.     

Intrahepatic Injection of Sodium Pentobarbital as an Alternative to Intraperitoneal Injection for the Euthanasia of Rats (Rattus norvegicus)

The most commonly accepted method of rat euthanasia in North America is intraperitoneal injection of sodium pentobarbital (PB). However, misinjection can occur, and intraperitoneal PB may cause pain and distress. The objective of this study was to test an alternative method of euthanasia: intrahepatic injection of PB. A pilot study was conducted to develop a method of intrahepatic injections (evaluated using CT scans and test injections), followed by a full study comparing intraperitoneal (n = 14) and intrahepatic PB injections (n = 66) in adult rats. Full study outcomes were: 1) time from injection to loss of righting reflex (LORR), 2) time from injection to cessation of heartbeat (CHB), 3) number of failed euthanasia attempts, and 4) confirmation of successful intrahepatic injection or misinjection via necropsy. All injections were performed by a veterinary student. CT revealed that intrahepatic injections were feasible. Times (median [range]) to LORR and CHB were faster after successful intrahepatic injections (LORR, 3 s [1 to 5 s]; CHB, 8 s [2 to 242 s]) than after intraperitoneal injections (LORR, 89.5 s [73 to 110 s], CHB: 284.5 s [237 to 423 s]). The misinjection rate was higher with intrahepatic injections (59%) than with intraperitoneal injections (29%), but intrahepatic misinjection still resulted in fast and successful euthanasia (LORR, 29 s [1 to 96 s]; CHB, 216 s [12 to 330 s]), with the injectate distributed between the intraperitoneal and intrahepatic locations. The number of failed euthanasia attempts with intrahepatic injections was low (n = 2). Intrahepatic injections show potential as an alternative to intraperitoneal injections for rat euthanasia.

Large numbers of laboratory rats are killed after the completion of research projects worldwide. Currently, the only method that is classified as acceptable by both the AVMA and Canadian Council on Animal Care (CCAC) is an overdose of sodium pentobarbital (PB) via either intraperitoneal or intravenous injection.^3,7 Of these, the intraperitoneal route is more commonly used in rats as it is easier and faster to perform. However, intraperitoneal injection has 2 important disadvantages that interfere with whether ‘euthanasia’ (“a good death”) is always achieved. The first disadvantage is an inherent misinjection rate that varies between 6% to 20% for rats.^{5,9,11,12,21,22,27,28,33} The consequences of a misinjection is a delay or failure to achieve loss of consciousness or death.^8,30,32,33 In these cases, the injection may need to be repeated or an alternative killing method applied.²⁰The second disadvantage is the potential for pain and distress. Converging evidence for this drawback includes elevated plasma corticosterone, tachycardia, hyperthermia, expression of immediate early response genes, electroencephalographic changes, visible signs of inflammation and behavioral changes, which have all been associated with intraperitoneal injection of PB.^{4,10,17,19,23,25,26,29} The source of nociception and potential pain is mostly due to alkaline pH (typically 10 to 12) of PB solution.²⁵The CCAC and AVMA euthanasia guidelines acknowledge that intraperitoneal injections may be painful and suggest the addition of a local anesthetic, such as lidocaine. However, neuronal and behavioral studies suggest that the addition of lidocaine does not eliminate nociception and pain.^2,18,29 Furthermore, the amount of lidocaine (or another buffer) that can be added is limited. As the pH descends below approximately 10, the PB precipitates.³² In addition, when misinjections occur, the potential for pain increases as a result of potential delay until loss of consciousness or pain resulting from the site of misinjection.²⁵These concerns indicate an ongoing need to refine the use of intraperitoneal PB or identify alternative methods to achieve euthanasia. A relatively unexplored injection route is intrahepatic injection. The current AVMA euthanasia guidelines describe intrahepatic injections as acceptable only in unconscious or anesthetized animals (with the exception of cats).³ This method has successfully been used in conscious cats,¹⁶ in which successful intrahepatic injection resulted in almost immediate recumbency, but intrahepatic injection of PB for euthanasia remains untested in rats. By contrast, intrahepatic injection in mice recently was shown to be unsuccessful.²¹We first performed a pilot study, with the objectives of: 1) identifying the appropriate injection site and angle and 2) testing the feasibility of the intrahepatic injection methods in rats. These results were applied in the main study, with the objective of investigating whether the intrahepatic injection technique could be an alternative to intraperitoneal injection for euthanasia in rats. We hypothesized that intrahepatic injection would result in a shorter time to loss of consciousness and death as compared with intraperitoneal injection, and would have fewer instances of failure to achieve death.     

寓科研于教学,提高药理学教学质量之刍议

药理学是联系药学和医学的重要桥梁学科,有科学研究参与的药理学教学能使枯燥的药理学课程变得生动有趣,其次科学研究的成果能使学生受到鼓舞,提高学生的学习兴趣。因此,结合科学研究的药理学教学方式有利于克服传统课堂教学存在的问题,提高教学质量;有利于高素质和创新型医药学科研人才的培养。