A method for multiparametric statistical quantification of the heterogeneity of free Na+ concentration by 19F MR spectroscopy: Proof of principle in silico and in vitro

Sodium(I) (Na⁺) is one of the most important cations in mammalian tissues. Since Na⁺ plays a key role in basic cell function, noninvasive methods for measuring intracellular concentrations of free sodium ions in biological tissue have been developed on the basis of ¹⁹F NMR spectroscopy. However, intracellular Na⁺ levels are often not uniform throughout a tissue volume (or voxel) being measured. In such cases, [Na⁺] heterogeneity is not reflected in results obtained by the classical technique, and may even result in biased average values. For this reason, we have designed an approach for quantifying [Na⁺] heterogeneity. First, the ¹⁹F MRS resonance from FCrown‐1 serving as a “Na⁺ probe” is transformed into a [Na⁺] curve. Then the digital points of the resulting [Na⁺] profile are used to construct a histogram with specially developed algorithms. From each [Na⁺] histogram, at least eight quantitative parameters describing the underlying statistical [Na⁺] distribution were computed: weighted median, weighted mean, standard deviation, range, mode(s), kurtosis, skewness, and entropy. In addition to our new paradigm, we present a first validation based on (i) computer simulations and (ii) experimentally obtained ¹⁹F MR spectra of model solutions. This basic proof of principle warrants future in vivo experiments, in particular because of its ability to provide quantitative information complementary to that made available by commonly used ²³Na MRI: (i) multiparametric statistical characterization of [Na⁺] distributions; (ii) total [Na⁺] heterogeneity analysis not intrinsically limited by the size of any MRI voxels; and (iii) analysis of unequivocally intracellular [Na⁺], as opposed to measurement of a combination of intra‐ and extracellular [Na⁺].     

A protocol for quantifying cardiogenic oscillations in dynamic 129Xe gas exchange spectroscopy: The effects of idiopathic pulmonary fibrosis

The spectral parameters of hyperpolarized ¹²⁹Xe exchanging between airspaces, interstitial barrier, and red blood cells (RBCs) are sensitive to pulmonary pathophysiology. This study sought to evaluate whether the dynamics of ¹²⁹Xe spectroscopy provide additional insight, with particular focus on quantifying cardiogenic oscillations in the RBC resonance. ¹²⁹Xe spectra were dynamically acquired in eight healthy volunteers and nine subjects with idiopathic pulmonary fibrosis (IPF). ¹²⁹Xe FIDs were collected every 20 ms (T_E = 0.932 ms, 512 points, dwell time = 32 μs, flip angle ≈ 20°) during a 16 s breathing maneuver. The FIDs were pre‐processed using the spectral improvement by Fourier thresholding technique (SIFT) and fit in the time domain to determine the airspace, interstitial barrier, and RBC spectral parameters. The RBC and gas resonances were fit to a Lorentzian lineshape, while the barrier was fit to a Voigt lineshape to account for its greater structural heterogeneity. For each complex resonance the amplitude, chemical shift, linewidth(s), and phase were calculated. The time‐averaged spectra confirmed that the RBC to barrier amplitude ratio (RBC:barrier ratio) and RBC chemical shift are both reduced in IPF subjects. Their temporal dynamics showed that all three ¹²⁹Xe resonances are affected by the breathing maneuver. Most notably, several RBC spectral parameters exhibited prominent oscillations at the cardiac frequency, and their peak‐to‐peak variation differed between IPF subjects and healthy volunteers. In the IPF cohort, oscillations were more prominent in the RBC amplitude (16.8 ± 5.2 versus 9.7 ± 2.9%; P = 0.008), chemical shift (0.43 ± 0.33 versus 0.083 ± 0.05 ppm; P < 0.001), and phase (7.7 ± 5.6 versus 1.4 ± 0.8°; P < 0.001). Dynamic ¹²⁹Xe spectroscopy is a simple and sensitive tool that probes the temporal variability of gas exchange and may prove useful in discerning the underlying causes of its impairment.     

Feasibility of precise and reliable glucose quantification in human whole blood samples by 1 tesla benchtop NMR

The standard procedure for blood glucose measurements is enzymatic testing. This method is cheap, but requires small samples of open blood with direct contact to the test medium. In principle, NMR provides non‐contact analysis of body fluids, but high‐field spectrometers are expensive and cannot be easily utilized under clinical conditions. Low‐field NMR systems with permanent magnets are becoming increasingly smaller and more affordable. The studies presented here aim at exploring the capabilities of low‐field NMR for measuring glucose concentrations in whole blood. For this purpose, a modern 1 T benchtop NMR spectrometer was used. Challenges arise from broad spectral lines, the glucose peak locations close to the water signal, low SNR and the interference with signals from other blood components. Whole blood as a sample comprises even more boundary conditions: crucial for reliable results are avoiding the separation of plasma and cells by gravitation and reliable reference values. First, the accuracy of glucose levels measured by NMR was tested using aqueous glucose solutions and commercially available bovine plasma. Then, 117 blood samples from oral glucose tolerance testing were measured with minimal preparation by simple pulse‐acquire NMR experiments. The analysis itself is the key to achieve high precision, so several approaches were investigated: peak integration, orthogonal projection to latent structure analysis and support vector machine regression. Correlations between results from the NMR spectra and the routine laboratory automated analyzer revealed an RMSE of 7.90 mg/dL for the best model. 91.5% of the model output lies within the limits of the German Medical Association guidelines, which require the glucose measurement to be within 11% of the reference method. It is concluded that spectral quantification of glucose in whole blood samples by high‐quality NMR spectrometers operating at 1 T is feasible with sufficient accuracy.     

乳腺癌组织内微血管分布的定量研究

用ＦⅧ相关抗原免疫组化染色定量观察４８例乳腺浸润癌。结果表明：有腋窝淋巴结转移组血管密度为每ｍｍ２１３５．４±４７．８个，无腋窝淋巴结转移组血管密度为每ｍｍ２９６．１±３１．４个，两组间的差异有显著意义（Ｐ＜０．０１）。同时血管的分布在癌组织旁每ｍｍ２１４２．４±４９．５个，在癌肿中央为每ｍｍ２９３．５±２８．６个，两组差异亦具有显著意义。结果揭示乳腺癌组织内血管生成与肿瘤的生长及腋窝淋巴结的转移关系密切。     

Quantitative analysis of HBV DNA level and HBeAg titer in hepatitis B surface antigen positive mothers and their babies: HBeAg passage through the placenta and the rate of decay in babies

It is well documented that perinatal transmission is the major cause of chronic HBV infection in China. However, the mechanisms of HBV perinatal transmission are not defined clearly. It is not known whether hepatitis B e antigen can cross the human placenta, and the rate of HBeAg decay in babies with and without HBV breakthrough has not been studied. In this study, HBV serological markers were investigated in 95 hepatitis B surface antigen positive pregnant women. These markers were also studied in the babies at birth and at the age of 6 months and 12 months. The data show that 7.4% (7/95) children were infected with HBV during the first year after birth despite receiving passive-active immunoprophylaxis with hepatitis B immune globulin and hepatitis B vaccine. The surface gene fragment of HBV DNA was cloned and sequenced following PCR amplification in 7 cases of HBsAg positive babies and their mothers. All babies had the same sequences as their mothers, although two babies also had sequences that would produce an amino acid substitution within the "a" determinant. Furthermore, we measured HBeAg titers and HBV DNA levels by using Abbott AxSYM system and LightCycler-based real-time fluorescence quantitative PCR in 54 mother-infant pairs. Thirty-three mothers were HBeAg positive, and 21 mothers were HBeAg negative. Seventy percent (23/33) of neonates from HBeAg-positive mothers were HBeAg positive at birth compared with 0% (0/21) of neonates from HBeAg negative mothers. HBeAg was present at higher titer in the birth sera of the babies with HBV breakthrough than in babies without breakthrough. HBeAg was cleared from the serum in all 19 babies without breakthrough. In 17 of these 19 babies, the HBeAg was cleared within 6 months, and in two babies clearance took 12 months. The mean serum HBV DNA level in the mothers of the 4 infants with HBV breakthrough was significantly higher than in the mothers of babies who did not become infected. In conclusion, this data suggests that HBeAg can cross the human placenta, and disappears from serum within 6 months in most babies. HBV DNA levels in hepatitis B carrier mothers are associated with the failure of HBIG and vaccine immunization, and the additional influence of transmitted HBeAg cannot be excluded.     

牡丹皮不同炮制品UPLC指纹图谱及多成分含量测定研究

目的 建立牡丹皮及其炮制品的超高效液相(UPLC)指纹图谱,并测定没食子酸、5-羟甲基糠醛(5-HMF)、对羟基苯甲酸、氧化芍药苷、芍药苷、苯甲酸、1,2,3,4,6-O-五没食子酰葡萄糖、丹皮酚等8个指标成分的质量分数,为牡丹皮药材质量标准的提高提供依据.方法 采用Zorbax Eclipse Plus-C18(2.1 mm×100 mm,1.8 μm)色谱柱,以质量分数0.1%甲酸水溶液-质量分数0.05%甲酸甲醇为流动相进行梯度洗脱,流速为0.2 mL/min,柱温为30 ℃,建立牡丹皮及其炮制品的UPLC指纹图谱,同时对以上8个成分进行测定.结果 建立了生丹皮、炒丹皮及丹皮炭3种药材的UPLC指纹图谱,以丹皮酚为参照峰,生丹皮和炒丹皮各标定了17个共有峰,丹皮炭标定了8个共有峰,除1批丹皮炭相似度为0.857外,样品与对照指纹图谱的相似度均大于0.944;除了苯甲酸的质量分数无明显变化外,其余化合物在牡丹皮炮制前后均有不同程度的增加或减少.结论 建立的UPLC指纹图谱及含量测定方法均具有较好的重现性,且分析时间短,方便快速,二者结合可直观地反映生丹皮、炒丹皮及丹皮炭3种药材之间成分的差异.     

Psychosocial factors affecting psychosomatic symptoms in Japanese schoolchildren

BACKGROUND: In a previous study we reported the prevalence of psychosomatic symptoms in 1619 Osaka elementary schoolchildren using our original general health questionnaire assessed by their parents. We found that psychosomatic symptoms were increasing with age. This study was designed to investigate psychosocial factors responsible for increasing psychosomatic symptoms with age. METHODS: We calculated a correlation coefficient between the physical complaints score (PCS) and these psychosocial variables using quantitation I of multiple regression analysis separately in the older (10-12 years, n = 860) and the younger age group (7-9 years, n = 759). RESULTS: A stronger relationship between PCS and psychosocial problems was found in the older (r = 0.719, P < 0.0001) than in the younger age group (r = 0.570, P < 0.0001). Further analysis demonstrated that difficulties in school performance and increasing psychological conflicts with human relationships were major causes of somatic complaints in the older age group. In the younger age group, however, immaturity of social skills seems to be a primary problem. In addition, poor parental interaction and the playing of computer games were found to increase somatic complaints in children. CONCLUSION: Children in the older age reflect the closer bio-psycho-socio interaction compared with their younger counterparts. This may give rise to a high incidence of psychosomatic disorders and school refusal in Japanese children. We emphasize that psychological support by parents in daily life is necessary to reduce psychosomatic symptoms in children.     

定量检测细胞凋亡的ELISA法

细胞凋亡普遍存在于胚胎发生及个体发育的多种生理和病理过程中,本文介绍了细胞凋亡,定量检测ＢＬＩＳＡ法的原理,方法及有关注意事项。     

对正常及大细胞性改变的肝细胞、肝癌和癌旁改变肝细胞ICM-DNA定量研究

目的探索正常肝细胞、大细胞性改变肝细胞、肝癌和癌旁改变肝细胞的DNA含量和形态特点.方法采用DNA组织化学染色和图像细胞分析术.结果10例癌旁改变(PC)组DNA指数(1.99±0.34)分别高于4例正常肝细胞(NH)组(0.98±0.12)和8例肝细胞的大细胞性改变(LCC)组(1.46±0.19)(P＜0.01),DNA倍体分布与NH、LCC和肝细胞癌(HCC)组存在差异(P＜0.05),核面积和核周长(74.0±52.3,63.9±33.9)低于26例HCC组(95.6±95.2,76.3±62.9)(P＜0.01).PC组中80%(8/10)为非整倍体DNA直方图表型,高于HCC组的53%(14/26),而多倍体占20%(2/10)低于HCC组26.92%(7/26)(P＜0.05).PC肝细胞核异型,核质浓缩,胞质嗜酸性或嗜碱性与HCC有过渡,置换性生长方式又与HCC相同.结论呈片块状分布或腺瘤样排列的PC肝细胞,其异常DI、非整倍体分布和DNA直方图表型应作为癌前状态.