122例酒精性肝病患者的临床研究

目的探讨酒精性肝病患者的治疗方法与临床疗效。方法回顾性分析我院收治的122例酒精性肝病患者的临床资料,所有患者随机分为2组,实验组61例患者在常规治疗的基础上采用阿托莫兰联合易善复进行治疗,对照组61例患者采用常规治疗的方法进行治疗。治疗结束后观察2组的临床疗效。结果实验组的疗效明显高于对照组,2组差异有统计学意义（P〈0.05）。结论阿托莫兰联合易善复治疗酒精性肝病疗效肯定,能明显改善酒精性肝病患者的临床症状,并促进黄疸的消退,促进肝功能恢复。     

[N‐Methyl‐11C]choline by on‐column reaction: a study on [11C]CH3I incorporation and the residual amount of precursor in the product

[N‐Methyl‐¹¹C]choline has been synthesized at room temperature by the reaction of [¹¹C]CH₃I with 2‐dimethylaminoethanol (DMAE), with the latter directly loaded on a weak cation‐exchange cartridge. Most of the efforts have been directed to reduce the amount of residual precursor in the product's final solution in order to make this tracer more suitable to brain studies. In the process, radiochemical yields and residual DMAE have been placed in relation with both the starting amount of precursor and the rinsing conditions used and compared with the more ‘traditional’ loading of the precursor on either a C18 cartridge or a loop. Comments and indications on the most convenient analytical technique and conditions for quantitative analysis, with particular emphasis on the precursor, are also reported. Under what we believe to be a fair compromise, [¹¹C]CH₃I incorporation yields of ca. 90% were easily achieved with a residual amount of starting material in the 8‐ to 12‐ppm range. Copyright © 2010 John Wiley & Sons, Ltd.     

Investigation of breast cancer using two‐dimensional MRS

Proton (¹H) MRS enables non‐invasive biochemical assay with the potential to characterize malignant, benign and healthy breast tissues. In vitro studies using perchloric acid extracts and ex vivo magic angle spinning spectroscopy of intact biopsy tissues have been used to identify detectable metabolic alterations in breast cancer. The challenges of ¹H MRS in vivo include low sensitivity and significant overlap of resonances due to limited chemical shift dispersion and significant inhomogeneous broadening at most clinical magnetic field strengths. Improvement in spectral resolution can be achieved in vivo and in vitro by recording the MR spectra spread over more than one dimension, thus facilitating unambiguous assignment of metabolite and lipid resonances in breast cancer. This article reviews the recent progress with two‐dimensional MRS of breast cancer in vitro, ex vivo and in vivo. The discussion includes unambiguous detection of saturated and unsaturated fatty acids, as well as choline‐containing groups such as free choline, phosphocholine, glycerophosphocholine and ethanolamines using two‐dimensional MRS. In addition, characterization of invasive ductal carcinomas and healthy fatty/glandular breast tissues non‐invasively using the classification and regression tree (CART) analysis of two‐dimensional MRS data is reviewed. Copyright © 2008 John Wiley & Sons, Ltd.     

Morphological and physiological properties of enhanced green fluorescent protein (EGFP)‐expressing wide‐field amacrine cells in the ChAT‐EGFP mouse line

Mammalian retinas comprise a variety of interneurons, among which amacrine cells represent the largest group, with more than 30 different cell types each exhibiting a rather distinctive morphology and carrying out a unique function in retinal processing. However, many amacrine types have not been studied systematically because, in particular, amacrine cells with large dendritic fields, i.e. wide‐field amacrine cells, have a low abundance and are therefore difficult to target. Here, we used a transgenic mouse line expressing the coding sequence of enhanced green fluorescent protein under the promoter for choline acetyltransferase (ChAT‐EGFP mouse) and characterized a single wide‐field amacrine cell population monostratifying in layer 2/3 of the inner plexiform layer (WA‐S2/3 cell). Somata of WA‐S2/3 cells are located either in the inner nuclear layer or are displaced to the ganglion cell layer and exhibit a low cell density. Using immunohistochemistry, we show that WA‐S2/3 cells are presumably GABAergic but may also release acetylcholine as their somata are weakly positive for ChAT. Two‐photon‐guided patch‐clamp recordings from intact retinas revealed WA‐S2/3 cells to be ON‐OFF cells with a homogenous receptive field even larger than the dendritic field. The large spatial extent of the receptive field is most likely due to the extensive homologous and heterologous coupling among WA‐S2/3 cells and to other amacrine cells, respectively, as indicated by tracer injections. In summary, we have characterized a novel type of GABAergic ON‐OFF wide‐field amacrine cell which is ideally suited to providing long‐range inhibition to ganglion cells due to its strong coupling.     

Maternal choline supplementation differentially alters the basal forebrain cholinergic system of young‐adult Ts65Dn and disomic mice

Down syndrome (DS), trisomy 21, is a multifaceted condition marked by intellectual disability and early presentation of Alzheimer's disease (AD) neuropathological lesions including degeneration of the basal forebrain cholinergic neuron (BFCN) system. Although DS is diagnosable during gestation, there is no treatment option for expectant mothers or DS individuals. Using the Ts65Dn mouse model of DS that displays age‐related degeneration of the BFCN system, we investigated the effects of maternal choline supplementation on the BFCN system in adult Ts65Dn mice and disomic (2N) littermates at 4.3–7.5 months of age. Ts65Dn dams were maintained on a choline‐supplemented diet (5.1 g/kg choline chloride) or a control, unsupplemented diet with adequate amounts of choline (1 g/kg choline chloride) from conception until weaning of offspring; post weaning, offspring were fed the control diet. Mice were transcardially perfused with paraformaldehyde, and brains were sectioned and immunolabeled for choline acetyltransferase (ChAT) or p75‐neurotrophin receptor (p75^NTR). BFCN number and size, the area of the regions, and the intensity of hippocampal labeling were determined. Ts65Dn‐unsupplemented mice displayed region‐ and immunolabel‐dependent increased BFCN number, larger areas, smaller BFCNs, and overall increased hippocampal ChAT intensity compared with 2N unsupplemented mice. These effects were partially normalized by maternal choline supplementation. Taken together, the results suggest a developmental imbalance in the Ts65Dn BFCN system. Early maternal‐diet choline supplementation attenuates some of the genotype‐dependent alterations in the BFCN system, suggesting this naturally occurring nutrient as a treatment option for pregnant mothers with knowledge that their offspring is trisomy 21. J. Comp. Neurol. 522:1390–1410, 2014. © 2013 Wiley Periodicals, Inc.     

Non‐invasive in vivo imaging of early metabolic tumor response to therapies targeting choline metabolism

The cholinic phenotype, characterized by elevated phosphocholine and a high production of total‐choline (tCho)‐containing metabolites, is a metabolic hallmark of cancer. It can be exploited for targeted therapy. Non‐invasive imaging biomarkers are required to evaluate an individual's response to targeted anticancer agents that usually do not rapidly cause tumor shrinkage. Because metabolic changes can manifest at earlier stages of therapy than changes in tumor size, the aim of the current study was to evaluate ¹H‐MRS and diffusion‐weighted MRI (DW‐MRI) as markers of tumor response to the modulation of the choline pathway in mammary tumor xenografts. Inhibition of choline kinase activity was achieved with the direct pharmacological inhibitor H‐89, indirect inhibitor sorafenib and down‐regulation of choline‐kinase α (ChKA) expression using specific short‐hairpin RNA (shRNA). While all three strategies significantly decreased tCho tumor content in vivo, only sorafenib and anti‐ChKA shRNA significantly repressed tumor growth. The increase of apparent‐diffusion‐coefficient of water (ADCw) measured by DW‐MRI, was predictive of the induced necrosis and inhibition of the tumor growth in sorafenib treated mice, while the absence of change in ADC values in H89 treated mice predicted the absence of effect in terms of tumor necrosis and tumor growth. In conclusion, ¹H‐choline spectroscopy can be useful as a pharmacodynamic biomarker for choline targeted agents, while DW‐MRI can be used as an early marker of effective tumor response to choline targeted therapies. DW‐MRI combined to choline spectroscopy may provide a useful non‐invasive marker for the early clinical assessment of tumor response to therapies targeting choline signaling.     

Brain extracellular space during spreading depression and ischemia

The change of extracellular space volume of rat brain cortex during ischemia and cortical spreading depression, CSD (Leāo4) was evaluated by a new method. The cortical surface was imgated with isotonic CSF containing the extracellular markers 50 mM cholin or 50 mM trimethyltris(hydroxymethyl)methyl ammonium ion (N-TRIS), and their extracellular concentrations were monitored by ion-selective microelectrodes. When steady-state for the concentration of these markers was attained, CSD evoked a reversible increase of the concentration of the markers, indicating shrinkage of the interstitial volume of distribution. During ischemia an initial slow rate of concentration increase was observed, followed a few minutes later by a rapid increase concomitant with the sharp rise in extracellular potassium concentration. During CSD and ischemia, the maximal increases of choline and N-TRIS concentration reflected a shrinkage of the extracellular space amounting to about 50% of the initial volume.     

Effects of acetazolamide on medullary extracellular pH and{\text{P}}_{{\text{CO}}_{\text{2}} } and on ventilation in peripherally chemodenervated catsand on ventilation in peripherally chemodenervated cats

Cyclic guanosine monophosphate (GMP) productions by alpha rat atrial natriuretic peptide 1–28 (-rANP), carbamylcholine or sodium nitroprusside were assessed in isolated glomeruli microdissected from collagenase-treated kidneys of 2- to 34-day-old and adult rats. In both young and adult animals, -rANP-stimulated cyclic GMP generation was proportional to the number of glomeruli and was enhanced in a dose-dependent and saturable fashion with increasing -rANP concentrations. The apparent activation constant values were 6.4 nM for 5-day-old and 9.7 nM for adult rats. Maximal doses of either -rANP or rANP 5–28 elicited similar responses in young and adult animals. Clear differences appeared between the developmental patterns of cyclic GMP productions stimulated by either -rANP, carbamylcholine or sodium nitroprusside. The response to -rANP was very large in the youngest rats tested, declined sharply during the suckling period and represented about 1.6 times the adult control level in 34-day-old rats. In contrast, the response to carbamylcholine was low after birth and rose progressively with age up to the adult level at the end of the weaning period, and the response to nitroprusside seemed to be independent of the animal's age.     

Expression of dopamine D2 receptor and choline acetyltransferase mRNA in the dopamine deafferented rat caudate-putamen

Summary In situ hybridization was used to study dopamine D2 receptor (D2R) and choline acetyltransferase (ChAT) mRNA expression in neurons of the rat forebrain, both on control animals and after a unilateral 6-hydroxydopamine (6-OHDA) lesion of midbrain dopamine neurons. D2R mRNA expressing neurons were seen in regions which are known to be heavily innervated by midbrain dopamine fibers such as caudate-putamen, nucleus accumbens and olfactory tubercle. ChAT mRNA expressing neurons were seen in caudate-putamen, nucleus accumbens and septal regions including vertical limb of the diagonal band. In caudate-putamen, approximately 55% of the medium sized neurons, which is the predominating neuronal cell-size in this region, were specifically labeled with the D2R probe. In addition, approximately 95% of the large size neurons in caudate-putamen were specifically labeled with both the D2R and ChAT probes, suggesting that most cholinergic neurons in the caudate-putamen express D2R mRNA. After a unilateral lesion of midbrain dopamine neurons, no change in the level of either D2R or ChAT mRNA were seen in the large size intrinsic cholinergic neurons in caudate-putamen. Similarily, no evidence was obtained for altered levels of D2R mRNA in medium size neurons in medial caudate-putamen, or nucleus accumbens. However, an increase in the number of medium size neurons expressing D2R mRNA was observed in the lateral part of the dopamine deafferented caudateputamen. Thus, it appears that midbrain dopamine deafferentation causes an increase in D2R mRNA expression in a subpopulation of medium size neurons in the lateral caudate-putamen.