Objective To construct a folic acid deficient model in zebrafish and to observe the axial development in folic acid deficient embryos, so as to probe the mechanism by which folic acid deficiency induces abnormal development of axis. Methods We constructed the folic acid deficient zebrafish model by both using the antagonism of dihydrofolate reductase (MTX) and knocking-down dihydrofolate reductase gene. Then we observed the axial excursion of folic acid deficient embryos at 17 hpf under microscope. We labeled and observed the positions of liver, spleen and heart by using whole-mount in situ hybridization with specific antisense RNA probes. The expressions of some genes, which are down stream factors of Nodal signal pathway and important for axial development, were detected by whole-mount in situ hybridization and Real-time PCR. Results Parts of folic acid deficient embryos had axial excursion and abnormal positions of liver, spleen and heart. The expressing intensities of ntl and gsc appeared normal in folic acid deficient embryos, but the expressing spatial patterns were abnormal, which revealed the malformation of axial mesoderm. Conclusions Folic acid deficiency induced the abnormal development of axis and the malformation of axial mesoderm. Folic acid deficiency had no obviouse effect on Nodal pathway.

Objective To observe the development of hematopoietic stem cell and the apoptosis of ICM(intermediate cell mass) in folic acid deficient zebrafish embryos and investigate the mechanism by which folic acid deficiency induces abnormal hematopoiesis.Method The folic acid deficient zebrafish model was induced by both using the dihydrofolate reductase antagonism methotrexate(MTX) and knock-down dihydrofolate reductase gene.The development of embryos was observed under microscope.The blood cells were detected by O-dianisidine staining.Whole-mount in situ hybridization and real-time PCR were performed to examine the expression of FLK-1,GATA1and GATA2.Apoptosis in intermediate cell mass(ICM) was examined by TUNEL(terminal-deoxynucleotidyl transferase mediated nick end labeling) method.Results The abnormal developments of ICMs were observed both in MTX treated embryos and DHFR knock-down embryos.O-dianisidine staining revealed that folic acid deficiency resulted in the decreasing number of blood cells.In folic acid deficient embryos,the expression of FLK-1、GATA1and GATA2 was reduced and the apoptosis in ICMs was increased.Conclusion The abnormal hematopoiesis in zebrafish induced by folic acid deficiency is related with the increasing apoptosis in ICMs and decreasing expressions of FLK-1,GATA1and GATA2.

Objective To study the putative mechanisms underlying fetal alcohol syndrome by comparative protein-profile analysis between normal and ethanol-treated zebrafish embryo with twodimensional electrophoresis (2-DE).Methods Zebrafish embryos were exposed in 400 mmol/L ethanol at dome stage for 3 hours,and then ethanol-induced abnormalities were observed.Proteomes of zebrafish embryos at early stages including zygote stage,dome stage,shield stage and 5-somite stage,were separated by 2-DE.The subtraction analysis method was applied to eliminate the interference from maternal derived proteins.The ethanol-treated embryos at 5-somite stage was analyzed by 2-DE,and the protein profile was compared with that generated from control embryos at the same stage.The data obtained from 2-DE analysis were verified by in-situ hybridization.Results 400 mmol/L ethanol treatment caused axial malformation (62%) and cyclopia (60%) in zebrafish embryos.The 2-DE analysis showed that the expression of Collagen2al (Col2a1) and TAR DNA binding protein (TDP) was decreased in 12 hours post fertilization (12 hpf) ethanol-treated embryos by 81% and 73%,respectively.The in-situ hybridization also demonstrated that the expression of Col2al in axial mesoderm was reduced by ethanol treatment at the same stage.But for 24 hpf ethanoltreated embryos,the expression of Col2al in axis recovered to a comparable level to that in control embryos,while the structure of neural tube was disrupted severely by ethanol exposure.Conclusions It is suggested that the expressions of Col2al and TDP were disrupted by ethanol during early stage,which might induce the zebrafish developmental abnormalities.The ethanol interference on early expression of Col2al is supposed to be one of the major reasons leading to later abnormalities of axis and neutral tube.

In the study, a quantum resonant cavity model based on wave-particle duality was proposed for the explanation of the dynamic processes of essence, vigor, and spirit in the human body in traditional Chinese medicine (TCM). It is assumed that there is a macro human order parameter (wave function), and its dynamics are governed by a macro potential field reflecting influences from heaven, earth, and society, and satisfy the generalized Schrodinger equation. This proposed model was applied in the study to interpret basic concepts of human body in TCM, with an aim to unfold the TCM development in the future.

Objective To study pulmonary wedge angiography ( PWA ) with hemodynamic the evaluation of children with congenital heart disease and pulmonary artery hypertension ( PAH) . Methods Hemodynamic measurement and pulmonary wedge angiography were performed in 50 children with congenital heart disease. Comparison and analysis were made from the data obtained from PWA and catheterization. Results After PWA, the patients were categorized into 3 groups according to the measured hemodynamics parameters:group A [ n=15, patients with normal mean pulmonary artery pressure ( mPAP≤25 mmHg) and normal pulmonary vessel resistance (PVR﹤300 dyne?s?cm5)], group B [n=24, patients with PAH (mPAP﹥25 mmHg) but normal PVR] and group C (n=11, patients with PAH and elevated PVR (PVR≥300 dyne?s?cm5). Rote of tapering (ROT) was significant lower in group C than in group A and B (F=42. 559,P﹤0. 05). Pulmonary circulation time (PCT) was higher in group C than in group A and B (F=6. 037,P﹤0. 05). ROT correlated negatively with PVR (r = -0. 606, P ﹤0. 05). PCT index correlated positively with PVR (r=0. 783,P=0. 01). There was no significant correlation between PCT and mean pulmonary artery hypertension (mPAP). Conclusions PWA may help to make quantitative analysis of the pulmonary vascular status in patients with congenital heart disease.

Methods: This paper introduces the concept of “order parameters” and proposes a method for establishing an order parameter model of gas discharge visualization (GDV) based on the principle of “mastering both permanence and change (MBPC)”. The method involved the following three steps. First, average luminous intensity () and average area () of the GDV images were calculated to construct the phase space, and the score of the health questionnaire was calculated as the health deviation index (H). Second, the k-means++ clustering method was employed to identify subclasses with the same health characteristics based on the data samples, and to statistically determine the symptom-specific frequencies of the subclasses. Third, the distance (d) between each sample and the “ideal health state”, which determined in the phase space of each subclass, was calculated as an order parameter describing the health imbalance, and a linear mapping was established between the d and the H. Further, the health implications of GDV signals were explored by analyzing subclass symptom profiles. We also compare the mean square error (MSE) with classification methods based on age, gender, and body mass index (BMI) indices to verify that the phase space possesses the ability to portray the health status of the human body. Results: This study preliminarily tested the reliability of the order parameter model on data samples provided by 20 participants. Based on the discovered linear law, the current model can use d calculated by measuring the GDV signal to predict H (R2 > 0.77). Combined with the symptom profiles of the subclasses, we explain the classification basis of the phase space based on the pattern identification. Compared with common classification methods based on age, gender, BMI, etc., the MSE of phase space-based classification was reduced by an order of magnitude. Conclusion In this study, the GDV order parameter model based on MBPC can identify subclasses and characterize individual health levels, and explore the TCM health meanings of the GDV signals by using subjective-objective methods, which holds significance for establishing mathematical models from TCM diagnosis principles to interpret human body signals.

Following the quantum theory-based physical model of the human body, a new interpretation of the traditional Chinese medicine (TCM) principle of “Cunkou reads viscera” is presented. Then, a Gaussian pulse wave model as a solution to the Schrodinger equation is shown to accurately describe 19 different pulse shapes, and to quantitatively capture the degree of Yin-Yang attributes of 13 pulse shapes. Furthermore, the model suggests using pulse depth and strength as leading-order quantity and pulse shape as first-order quantity, to characterize the hierarchical resonance between the human body and the environment. The future pulse informatics will focus on determining an individual’s unique quantum human equilibrium state, and diagnose its health state according to the pulse deviation from its equilibrium state, to truly achieve the high level of TCM: “knowing the normal state and reaching the change”.

Objective To evaluate the regulatory effect of Diwu Yanggan(DWYG)Decoction on lysoglycerophospholipids(Lyso-GPLs)in circulating exosomes in a mouse model of nonalcoholic fatty liver disease(NAFLD).Methods Circulating exosomes isolated from mouse serum by size exclusion chromatography were morphologically characterized using transmission electron microscope and examined for surface markers CD9,CD63 and TSG101 using Western blotting.Twenty-four male Kunming mice were randomized into 3 groups for normal feeding(control,n=8)or high-fat diet feeding for 1 week to induce NAFLD,after which the latter mice were given DWYG decoction(treatment group,n=8)or normal saline(model group,n=8)by gavage for 4 weeks.After the last treatment,blood samples were collected from the mice for testing serum TC,HDL-C,LDL-C,ALT and AST levels and isolating circulating exosomes.Using multivariate statistical analysis based on targeted metabolomics strategy,the potential biomarkers for Lyso-GPLs in the exosomes were screened.Results The isolated exosomes about 100 nm in size had a typical saucer-like structure with distinct double-layer membranes and a mean particle size of 137.5 nm and expressed the specific surface marker proteins CD9,CD63 and TSG101.The mouse models of NAFLD had significantly increased serum levels of TC,HDL-C,LDL-C and AST and lowered serum ALT level.A total of 43 Lyso-GPLs with significant reduction after DWYG Decoction treatment were identified in NAFLD mice.Conclusion DWYG Decoction can regulate Lyso-GPLs in circulating exosomes in NAFLD mice,which provides a new clue for studying the therapeutic mechanism of DWYG Decoction for liver disease.

Objective To evaluate the regulatory effect of Diwu Yanggan(DWYG)Decoction on lysoglycerophospholipids(Lyso-GPLs)in circulating exosomes in a mouse model of nonalcoholic fatty liver disease(NAFLD).Methods Circulating exosomes isolated from mouse serum by size exclusion chromatography were morphologically characterized using transmission electron microscope and examined for surface markers CD9,CD63 and TSG101 using Western blotting.Twenty-four male Kunming mice were randomized into 3 groups for normal feeding(control,n=8)or high-fat diet feeding for 1 week to induce NAFLD,after which the latter mice were given DWYG decoction(treatment group,n=8)or normal saline(model group,n=8)by gavage for 4 weeks.After the last treatment,blood samples were collected from the mice for testing serum TC,HDL-C,LDL-C,ALT and AST levels and isolating circulating exosomes.Using multivariate statistical analysis based on targeted metabolomics strategy,the potential biomarkers for Lyso-GPLs in the exosomes were screened.Results The isolated exosomes about 100 nm in size had a typical saucer-like structure with distinct double-layer membranes and a mean particle size of 137.5 nm and expressed the specific surface marker proteins CD9,CD63 and TSG101.The mouse models of NAFLD had significantly increased serum levels of TC,HDL-C,LDL-C and AST and lowered serum ALT level.A total of 43 Lyso-GPLs with significant reduction after DWYG Decoction treatment were identified in NAFLD mice.Conclusion DWYG Decoction can regulate Lyso-GPLs in circulating exosomes in NAFLD mice,which provides a new clue for studying the therapeutic mechanism of DWYG Decoction for liver disease.