Time-resolved quantification of the dynamic extracellular space in the brain during short-lived event: methodology and simulations.

Two macroscopic parameters describe the interstitial diffusion of substances in the extracellular space (ECS) of the brain, the ECS volume fraction α and the diffusion tortuosity λ. Past methods based on sampling the extracellular concentration of a membrane-impermeable ion tracer, such as tetramethylammonium (TMA+), can characterize either the dynamic α(t) alone or the constant α and λ in resting state but never the dynamic α(t) and λ(t) simultaneously in short-lived brain events. In this work, we propose to use a sinusoidal method of TMA+ to provide time-resolved quantification of α(t) and λ(t) in acute brain events. This method iontophoretically injects TMA+ in the brain ECS by a sinusoidal time pattern, samples the resulting TMA+ diffusion waveform at a distance, and analyzes the transient modulations of the amplitude and phase lag of the sampled TMA+ waveform to infer α(t) and λ(t). Applicability of the sinusoidal method was verified through computer simulations of the sinusoidal TMA+ diffusion waveform in cortical spreading depression. Parameter sensitivity analysis identified the sinusoidal frequency and the interelectrode distance as two key operating parameters. Compared with other TMA+-based methods, the sinusoidal method can more accurately capture the dynamic α(t) and λ(t) in acute brain events and is equally applicable to other pathological episodes such as epilepsy, transient ischemic attack, and brain injury. Future improvement of the method should focus on high-fidelity extraction of the waveform amplitude and phase angle. NEW & NOTEWORTHY An iontophoretic sinusoidal method of tetramethylammonium is described to capture the dynamic brain extracellular space volume fraction α and diffusion tortuosity λ. The sinusoidal frequency and interelectrode distance are two key operating parameters affecting the method's accuracy in capturing α(t) and λ(t). High-fidelity extraction of the waveform amplitude and phase lag is critical to successful sinusoidal analyses.

Time-resolved quantification of the dynamic extracellular space in the brain: study of cortical spreading depression.

Extracellular diffusion in the brain is customarily characterized by two parameters, the extracellular space (ECS) volume fraction α and the diffusion tortuosity λ. How these two parameters are temporarily modified and correlated in a physiological/pathological event remains unclear to date. Using tetramethylammonium (TMA+) as an ECS ion tracer in a newly updated iontophoretic sinusoidal method, we studied in this work the dynamic α(t) and λ(t) in rat somatosensory cortex during spreading depression (SD). Temporal variations of α(t) and λ(t), as evoked by SD, were obtained through analyses of the extracellular TMA+ diffusion waveform resulting from a sinusoidally modulated point source. Most of the time, cortical SD induced coordinated α(t) decreases and λ(t) increases. In rare occasions, SD induced sole decreases of α(t) with no changes in λ(t). The independent modulation of α(t) and λ(t) was neither associated with cortical anatomy nor with the specific shape of the SD field potential wave. Changes of α(t) and λ(t) often took place acutely at the onset of SD, followed by a more transient modulation. Compared with the prior iontophoretic methods of TMA+, the sinusoidal method provides time-resolved quantification of α(t) and λ(t) in relative terms but also raises a higher property requirement on the TMA+-selective microelectrode. The sinusoidal method could become a valuable tool in the studies of the dynamic ECS response in various brain events. NEW & NOTEWORTHY An iontophoretic sinusoidal method was applied to study the dynamic changes of two extracellular space parameters, the extracellular volume fraction α(t) and tortuosity λ(t), in the brain during cortical spreading depression. Both parameters showed coordinated (most often) and independent (rarely) modulations in spreading depression. The sinusoidal method is equally applicable to other acute pathological events and a valuable tool to study the functional role of extracellular space in brain events.

Test of the Equivalence Principle with Chiral Masses Using a Rotating Torsion Pendulum.

Here we present a new test of the equivalence principle designed to search for the possible violation of gravitational parity using test bodies with different chiralities. The test bodies are a pair of left- and right-handed quartz crystals, whose gravitational acceleration difference is measured by a rotating torsion pendulum. The result shows that the acceleration difference towards Earth Δa_{left-right}=[-1.7±4.1(stat)±4.4(syst)]×10^{-15} m s^{-2} (1-σ statistical uncertainty), correspondingly the Eötvös parameter η=[-1.2±2.8(stat)±3.0(syst)]×10^{-13}. This is the first reported experimental test of the equivalence principle for chiral masses and opens a new way to the search for the possible parity-violating gravitation.

[Intestinal absorption and metabolism of Puerariae Lobatae Radix decoction by in situ closed-loop method].

Biopharmaceutics classification system of Chinese materia medica (CMMBCS) emphasizes characteristic of the multi-component environment based on the drug solubility and permeability. In this study, the in situ closed-loop method combined with LC-MS technique was utilized to study the intestinal absorption and metabolism of Puerariae Lobatae Radix decoction (PLRD), providing selection basis for intestinal permeability components in CMMBCS. A total of 36 components were identified from PLRD. Among them, 17 components could be detected in the plasma sample, indicating that 17 components could be absorbed into blood, so these 17 components could be used as intestinal permeability evaluation components in CMMBCS. The other 19 components were not detected in the plasma sample, suggesting that they may not be absorbed or metabolized by the gut wall enzymes.

[A correlation research on Chinese medical syndromes of chronic heart failure and various complications].

OBJECTIVE: To study the correlation between various complications of chronic heart failure (CHF) patients and Chinese medical syndromes, thus indicating distribution laws of Chinese medical syndromes in various complications of CHF patients. METHODS: Chinese medical syndrome typing was performed in 630 CHF patients by cross-sectional study of the demographic data, history of present diseases, related information on Chinese medical four diagnostic methods, and the distribution of complications. Logistic regression analysis was used to determine the correlation of various complications of CHF patients and Chinese medical syndromes. RESULTS: In this study, recruited were common complications such as hypertension, diabetes, arrhythmia, hyperlipemia, and cerebral vascular accident, and so on. Main syndromes were sequenced as qi deficiency syndrome, blood stasis syndrome, water retention syndrome, yin deficiency syndrome, phlegm turbid syndrome, yang deficiency syndrome. Results of Logistic regression analysis indicated that correlation existed between common complications and Chinese medical syndromes. In CHF complicated hypertension patients, Logistic regression analysis showed qi deficiency syndrome and yang deficiency syndrome were negatively correlated with hypertension (P < 0.05). In CHF complicated diabetes patients, Logistic regression analysis showed phlegm turbid syndrome and water retention syndrome were positively correlated with diabetes (P < 0.05). In CHF complicated arrhythmia patients, there was no statistical difference in the distribution of each syndrome (P > 0.05). In CHF complicated hyperlipemia patients, Logistic regression analysis showed qi deficiency syndrome and water retention syndrome were negatively correlated with hyperlipemia (P < 0.05), while blood stasis syndrome, yin deficiency syndrome, and phlegm turbid syndrome were positively correlated with hyperlipemia (P < 0.01). In CHF complicated cerebral vascular accident patients, Logistic regression analysis showed qi deficiency syndrome and yang deficiency syndrome were negatively correlated with cerebral vascular accident (P < 0.01, P < 0.05). CONCLUSIONS: There existed certain correlations between complications of CHF and the distribution of main Chinese medical syndromes. It could be used as guidance for treating CHF and its various complications by Chinese medicine and pharmacy.

Transcriptome analysis revealed enrichment pathways and regulation of gene expression associated with somatic embryogenesis in Camellia sinensis.

The high frequency, stable somatic embryo system of tea has still not been established due to the limitations of its own characteristics and therefore severely restricts the genetic research and breeding process of tea plants. In this study, the transcriptome was used to illustrate the mechanisms of gene expression regulation in the somatic embryogenesis of tea plants. The number of DEGs for the (IS intermediate stage)_PS (preliminary stage), ES (embryoid stage)_IS and ES_PS stages were 109, 2848 and 1697, respectively. The enrichment analysis showed that carbohydrate metabolic processes were considerably enriched at the ES_IS stage and performed a key role in somatic embryogenesis, while enhanced light capture in photosystem I could provide the material basis for carbohydrates. The pathway analysis showed that the enriched pathways in IS_PS process were far less than those in ES_IS or ES_PS, and the photosynthesis and photosynthetic antenna protein pathway of DEGs in ES_IS or ES_PS stage were notably enriched and up-regulated. The key photosynthesis and photosynthesis antenna protein pathways and the Lhcb1 gene were discovered in tea plants somatic embryogenesis. These results were of great significance to clarify the mechanism of somatic embryogenesis and the breeding research of tea plants.

[The FTIR spectral characteristics and comparison study of Astragalus menbranceus soil].

To study the genuine soil of Astragalus menbranceus grows, FTIR spectrometry was used, which is accurate, simple and efficient and has high resolution. The genuine soils include six areas in Hunyuan of Shanxi province, three areas in Yingxian of Shanxi province, Fansi of Shanxi province, and Guyang of Inner Mongolia. Different growth years of two to five for each area were also studied. The results show that there are significant differences between Astragalus menbranceus soil FTIR spectrometry and general soil's, between soil of Astragalus menbranceus growth and radix codonopsitis growth, between different soil of Astragalus menbranceus growth, providing useful information for the area chose of Chinese herb cultural and transplantation.

Influence of the tilt error motion of the rotation axis on the test of the equivalence principle with a rotating torsion pendulum.

Improving the precision of current tests of the equivalence principle with a rotating torsion pendulum requires a more complete analysis of systematic effects. Here, we discuss in detail one of the important systematic effects, the influence from the tilt error motion of the rotation axis of a rotary stage, namely, wandering of the instantaneous rotation axis around its average direction. Its influence on the rotating torsion pendulum is modeled phenomenologically, and the parameters in the model are calibrated. It is shown that the influence can contribute a correction of η ≈ 5 × 10-13 to the equivalence-principle violating parameter for a rotary stage whose tilt error motion of interest is about 31 nrad in magnitude. We also show that such an influence can be reduced to the level of η ≈ 1 × 10-14 by means of active compensation of the tilt error motion using a set of piezoelectric actuators placed under the stage stator.

Clinical manifestations and acid alpha-glucosidase mutation characterisation of a cohort of patients with late-onset Pompe disease in eastern China.

BACKGROUND: Pompe disease is a rare, progressive, and life-threatening autosomal recessive disorder. In its late-onset form, the disease is primarily characterised by mild progressive proximal limb and respiratory muscle weakness. Mutations in the acid alpha-glucosidase (GAA) gene cause lysosomal enzyme GAA to be significantly reduced or missing altogether, for which supplementation can be given through enzyme replacement therapy. METHODS: Fourteen patients diagnosed with late-onset Pompe disease (LOPD) in the First Affiliated Hospital of Nanjing Medical University from 2017 to 2021 were enrolled. GAA activity was measured based on enzymatic activity in dried blood spots, and next-generation sequencing was used to detect mutations in the GAA gene. The impacts of novel missense variants were determined by five different prediction algorithms. The structural figures of novel variants and their wide types were processed with PyMOL. RESULTS: The study included 14 patients with LOPD (male-to-female ratio, 1:1) from eastern China. The median age at symptom onset and diagnosis was 15.0 years (7-36 years) and 21.5 years (8-47 years), respectively. The median diagnostic delay from onset was 3.0 years (0-22 years). Proximal muscle weakness was the first prominent symptom in 8 patients, while the other 6 patients experienced respiratory failure, chest congestion and asthma, and scoliosis. The most frequent mutation of the GAA gene was c.2238G>C (p.W746C), which was observed at an allele frequency of 14.3% (4/28) and in 28.6% of patients (4/14). Four novel variants potentially related to the pathogenicity of LOPD were found: c.1299G>C (p.Q433H), c.1409A>G (p.N470S), c.2242delG (p.E748Rfs*16), and c.2832delA (p.E945Sfs*78). CONCLUSIONS: The c.2238G>C (p.W746C) mutation was the most common mutation in 14 patients with LOPD from eastern China. This study has identified four novel variants in patients with LOPD. Predicting the pathogenicity of these novel variants may increase the understanding of the genetic mutation spectrum in LOPD. Our findings may also improve recognition of the characteristics of Chinese patients with LOPD.

[Gel electrophoresis analysis on plasma differential protein in patients with unstable angina of blood-stasis pattern].

OBJECTIVE: To seek the plasma differential proteins in patients with unstable angina of blood-stasis pattern (UA-BSS) for exploring the proteomic specialty in them by way of two-dimensional difference gel electrophoresis (DIGE) detection on plasma of patients and healthy persons. METHODS: Using DIGE and tandem mass spectrometry, comparative proteomic study was conducted on the plasma of 12 UA patients of qi-deficiency and blood-stasis pattern (UA-QBS), 12 UA patients of phlegm-stasis cross-blocking pattern (UA-PSS) and 12 healthy volunteers. RESULTS: Preliminary results showed that Haptoglobin beta chain, DBP, HBB, HBA, Transthyretin, ApoA- I, ApoA-IV were significantly differentially expressed in both patterns, while Haptoglobin alpha1 chain, alpha-1-acid glycoprotein, ApoC-III, ApoA-II, ApoC-II, ApoJ, and Haptoglobin alpha 2 chain were only seen differentially expressed in the UA-PSS patients, alpha1-antitrypsin, Fibrinogen gamma chain, and Fibrin beta were only seen differentially expressed in UA-QBS patients. CONCLUSION: The common proteomics characteristics of patients of QBS and PSS patterns may be correlated with inflammatory reaction and metabolic disturbance (including blood lipid and blood oxygen).

[Edaphic FTIR finger print and the area choice of Chinese herb culture].

UNLABELLED: In order to search for the difference of the soil of Chinese herb growth, and provide useful information for the area choice of Chinese herb culture, 53 soil samples were detected by FTIR. The results showed that all the samples have 8 peaks at 3,621, 3,425, 1,642, 1,435, 1,029, 878, 760 and 690 cm(-1), respectively, but the relative intensity is different. In addition, the peaks at 1,796, 2,982 and 2,874 cm(-1) show difference too. This can be used to distinguish soil in different area. The study also showed that soil in Tianshui and Dingxi of Gansu province are similar to that in Shanxi province. CONCLUSIONS: FTIR finger print of soil can provide useful information for the area choice of Chinese herb culture and transplantation and can be used in the study of soil.

[Study on Chinese herb Astragalus membranceus by FTIR fingerprint].

UNLABELLED: The aim of the study is to find new approach to identifying different growth years and different producing areas of astragalus membranceus simply and rapidly. The FTIR spectrometry, which is accurate, simple and efficient and has high resolution, was used to obtain the sample's FTIR spectrometry. After simple separation and extraction, the drying powder of the sample was detected. In 22 samples, there were 9 common peaks for identifying the authenticity of astragalus membranceus and 5 different peaks. According to the number, shape, and relative intensity of the peak, different growth years and different producing areas of astragalus membranceus can be distinguished. CONCLUSION: It is convenient and fast to identify different growth years and different producing areas of astragalus membranceus by FTIR fingerprint.

[Investigative strategy for research on biological basis of traditional Chinese medicine syndrome: feature selection-based data mining methods].

This paper is devoted to discussing two research patterns of biological basis of traditional Chinese medicine syndrome and presenting a research strategy for data mining methods. It points out that data mining methods which are based on feature selection are better fit for investigating biological basis of traditional Chinese medicine syndrome. Based on such a discussion, the concept of "characteristic pattern" is proposed to bridge the gap between "golden index" and biological basis of traditional Chinese medicine syndrome. This paper presents a novel research avenue for investigating biological basis of traditional Chinese medicine syndrome.

[Diagnosis and treatment rule of traditional Chinese medicine for syndrome factors of chronic congestive heart failure: a study based on Shannon entropy method].

OBJECTIVE: The primary aim of this research is to systematically sort out and analyze available clinical documents for chronic congestive heart failure in traditional Chinese medicine (TCM) and integrated traditional Chinese and Western medicine, and to explore the diagnosis and treatment rule of TCM syndrome factors with data mining method. METHODS: Based on the China National Knowledge Internet (CNKI) and Chinese Biomedical Literature Database (CBM), using "chronic heart failure" or "chronic congestive heart failure" or "chronic cardiac insufficiency" and "traditional Chinese medicine" or "syndrome" or "integrated traditional Chinese and Western medicine" as keywords for literature selection, we used Shannon entropy nonlinear complex system method for the feature extraction after data preprocessing. RESULTS: The statistics showed that yang deficiency, qi deficiency, blood stasis, water retention, yin deficiency, and turbid phlegm were principal TCM syndrome factors; herbs of qi-tonifying, blood-activating, water-draining, phlegm-dispelling, exterior-releasing, interior-warming were main drugs for clinical application. The symptoms and Chinese herbs which intimately connect with TCM syndrome factors were extracted, while their diagnostic contribution degrees were quantified by correlation coefficient. The symptoms which had high scores were general syndromes while the symptoms which had low scores were specific manifestations of a certain patient. Common Chinese herbs corresponding to each syndrome factor were picked out, indirectly reflecting their frequencies in clinical application. CONCLUSION: Based on data mining of the relationship among the four diagnostic methods of TCM, syndrome factors and herbs, this study provided references for differentiation of TCM syndrome and selection of Chinese medicine according to clinical syndrome factors.

[Study on proteomic specificity of unstable angina with qi deficiency and blood stasis syndrome].

OBJECTIVE: To seek the special plasma molecule in unstable angina (UA) patients of qi deficiency and blood stasis syndrome (QDBS) and method for explore the proteomic specificity of the disease. METHODS: LC-MS(E) analysis was performed in UA patients of QDBS or non-QDBS and in healthy persons after the 6 proteins with optimal abundance in plasma being removed by polyclonal antibody affinity column (product of Agilent Co. USA). RESULTS: Actin were found only expressed, and FN, ApoH and ANXA6 were found highly expressed in plasma of patients with UA-QDBS, suggesting they might be the special molecules for the disease. Moreover, as compared with health persons, SAA, CP, MYH11 and C6 showed high expression, and the 6 proteins, e.g. A1BG, ApoA4, GSN, HBB, HBD and TF, showed low expression in the plasma of UA-QDBS patients. CONCLUSION: UA-QDBS might belong to a kind inflammatory reaction. There are simultaneous existence of myocardial injury, blood coagulation factor abnormality, lipid metabolic disorder and oxygen transport obstacle in patients of UA-QDBS, they influence and interact mutually. The newly discovered differential proteins might provide clues for studying or discovering new protein targets of angina relieving drugs. The new technique of label free quantitative proteomics is an efficient method for bio-marker research of diseases and syndromes.

[Differentially expressed protein study on unstable angina blood-stasis syndrome by fluorescence labelling method].

Fluorescence labelling method was used seeking differentially expressed plasma proteins of unstable angina blood-stasis syndrome. Three fluorescence labelling dyes (Cy2, Cy3 and Cy5) were used on two classes of species (plasma of 12 patients with unstable angina blood-stasis syndrome and 12 healthy volunteers). Two dimensional gel electrophoresis was carried out, and three different fluorescence scans were performed. By analyzing the graphs and mass spectrometry, the differentially expressed plasma proteins of unstable angina were obtained. Blood-stasis syndrome. Results show that fibrinogen beta chain, fibrinogen gamma chain, alpha1-antitrypsin, haptoglobin beta chain and haptoglobin alpha2 chain were significantly highly expressed in the plasma of unstable angina blood-stasis syndrome patient, while ApoA-IV, ApoA-I and transthyretin were decreased in the plasma of unstable angina blood-stasis syndrome patient. These identified proteins could be divided into two categories according to their functions: (1) Acute phase reactive protein; (2) Apolipoprotein. In conclusion, fibrinogen beta chain, fibrinogen gamma chain, alpha1-antitrypsin, transthyretinc, haptoglobin beta chain, ApoA-IV, ApoA-I and haptoglobin alpha2 chain are differentially expressed in the patients with unstable angina blood-stasis syndrome and control group, some differentially expressed proteins were correlated with inflammatory reaction or lipid metabolic disorder, and these proteins could provide clues to the study and discovery of new protein targets for antianginal drugs. The fluorescence labelling method is suitable for discovery of differentially expressed proteins.

Loss accounting of environmental pollution within Pearl River Delta region, South China.

Economic development puts pressure on environment through air, water and land degradation, which in turn brings real costs to the economy. Actual economy growth should therefore consider the environmental degradation cost resulted from economic activities. Pearl River Delta (PRD) region as a typical delta area with rapid development in China, is with great significance to understand the loses resulted from environmental pollution. This study conducts an environmental economic accounting within the PRD region from 2011 to 2015 using the environmental degradation cost accounting approach. We identified and calculated the economic, agricultural, industrial and social losses resulting from air, water and waste pollution with different valuation methods, which includes shadow price, replacement costs, market value method, etc. The results showed the total environmental degradation cost ranged from 18.1 to 19.8 billion US$ and the environmental degradation index declined slightly over the years, with significant differences among cities. It implied that the environmental condition of PRD region has been continuously improved over the years, but the capacity of environment control between cities had large differences. Cities in PRD region should therefore take measures tailored to their current situation to optimize their resource endowment and industrial structure, to overcome the conflicts between economic development and environmental protection. For cities with relatively high degradation cost, it is urgent to accelerate the efforts in improving the quality of the environment and ecosystem. For cities with lower degradation cost, it is important to take actions to keep on a sustainable and ecological efficient developing path. MAIN FINDINGS: The total environmental degradation cost of the PRD region is firstly calculated with insights on environmental management.

[Effects of tetrandrine on nuclear factor-kappaB expression in leukemia multidrug-resistant cell line K562/A02].

OBJECTIVE: To investigate the effects of tetrandrine (Tet) on nuclear factor kappaB (NF-kappaB) expression in leukemia cell line K562 and multidrug-resistant K562/A02 cell line. METHODS: The activations of NF-kappaB in K562 and K562/A02 cells and the effects of 1 micromol/L Tet on NF-kappaB expression were determined by immunocytochemistry and Western blot assay. RESULTS: Tet had no effect on NF-kappaB expression in K562 cells after 6- and 12-hour treatment (P>0.05), and K562/A02 cells displayed higher level of NF-kappaB protein expression than their parental K562 cells (P<0.01). Tet could significantly down-regulate NF-kappaB protein expression and nuclear translocation in K562/A02 cells shown by immunocytochemistry and Western blot, and this decrease became more significant after 12-hour treatment than after at 6-hour treatment (P<0.05). CONCLUSION: Activation of NF-kappaB may be related to multidrug resistance of K562/A02 cell line. And the inhibition of NF-kappaB activation by Tet leads to multidrug resistance reversal in K562/A02 cell line.