Characterising genes associated with flowering time in carrot (Daucus carota L.) using transcriptome analysis.

Carrot is generally regarded as a biennial plant with an obligatory vernalization requirement. Early spring cultivation makes plants vulnerable to premature bolting, which results in a loss of commercial value. However, our knowledge of flowering time genes and flowering mechanisms in carrot remain limited. Bolting behavior of D. carota ssp. carota 'Songzi', a wild species sensitive to flower induction by vernalization and photoperiod, and orange cultivar 'Amsterdam forcing', and their offspring were investigated in different growing conditions. We performed RNA-seq to identify the flowering time genes, and digital gene expression (DGE) analysis to examine their expression levels. The circadian patterns of related genes were identified by qPCR. The results showed bolting behavior of carrot was influenced by low temperature, illumination intensity and photoperiod. A total of 45 flowering time-related unigenes were identified, which were classified into five categories including photoperiod, vernalization, autonomous and gibberellin pathway, and floral integrators. Homologs of LATE ELONGATED HYPOCOTYL (LHY) and CONSTANS-LIKE 2 (COL2) were more highly expressed under short day condition than under long day condition. Homologs of COL2, CONSTANS-LIKE 5 (COL5), SUPPRESSION OF OVEREXPRESSION OF CONSTANS 1 (SOC1), FLOWERING LOCUS C (FLC) and GIBBERELLIC ACID INSENSITIVE (GAI) were differentially expressed between 'Songzi' and 'Amsterdam forcing'. The homolog of COL2 (Dct43207) was repressed by light, but that of COL5 (Dct20940) was induced. A preliminary model of genetic network controlling flowering time was constructed by associating the results of DGE analysis with correlation coefficients between genes. This study provides useful information for further investigating the genetic mechanism of flowering in carrot.

Caveolin-1 redistribution in human endothelial cells induced by laminar flow and cytokine.

Caveolin-1 is a principal component of caveolae and is involved in signaling transduction in a number of cells. A hypothesis was proposed in this work that mechanical forces due to flow induce caveolin-1 translocation. So the changes of caveolin-1 expression and distribution in cultured endothelial cells (HUVECs) exposed to a steady laminar flow were studied. For comparing with the influence of cytokine, caveolin-1 in the cells stimulated by TNF-alpha was also investigated. Indirect immunofluorescence and double fluorescence labeling showed that in control cells, caveolin-1 was primarily localized on the cell surface, which corresponded to the peripheral distribution of F-actin, and presented some local concentrations. In the cells exposed to a laminar flow (1.0 Pa), caveolin-1 distribution showed a time-dependent variation. After 24 h of shear, the local concentration of caveolin-1 was found, in the most cells, at upstream side of cell body. Also more caveolin-1 molecules were observed in the cells. In contrast, TNF-alpha induced a decrease of caveolin-1 in cells. The redistribution of caveolin-1 seems to be correlated to F-actin organization.

Remodelling of zero-stress state of small intestine in streptozotocin-induced diabetic rats. Effect of gliclazide.

BACKGROUND: Biomechanical properties in terms of residual strains in diabetic small intestine have not been studied. Furthermore, no data have been reported on affect of gliclazide on gastrointestinal complications of diabetes. AIMS: To determine remodelling of zero-stress state of small intestine in streptozotocin-induced diabetic rats and effect of gliclazide treatment. MATERIALS: Morphological properties and residual strains were studied in duodenum, jejunum and ileum obtained from diabetic rats, gliclazide-treated diabetic rats and normal rats (n = 8 each group). METHODS: Diabetes was induced by single intraperitoneal injection of 65 mg/kg streptozotocin. Gliclazide (10 mg kg(-1) day(-1) was injected directly into stomach lumen by intragastric gavage twice daily. Experimental period was 35 days. To approach no-load state; intestinal segments were surgically excised and cut transversely into short ring-shaped segments. Each ring was cut radially to obtain geometry of zero-stress state. Circumferential length, the wall thickness and opening angle were measured from digital images of each specimen and residual strains were computed. RESULTS: Blood glucose level of diabetic group (approximately 20 mmol/l) was consistently higher than that in normal group (approximately 4 mmol/l) after induction of diabetes (p < 0.001). Gliclazide lowered average blood glucose level to between 10 and 15 mmol/l (p < 0.001). Plasma insulin levels of both diabetic groups (average between 10 and 15 pmol/l) were significantly lower than those in normal group (average approximately 18 pmol/l, p < 0.05). Wet weight per unit length and wall thickness of duodenum, jejunum and ileum were significantly higher in Diabetes group than those in Normal group (p < 0.05). Opening angle and absolute value of residual strain were significantly smaller in duodenum and larger in jejunum and ileum in Diabetes group than in Normal group (p < 0.001). Gliclazide treatment partly restored these changes (p < 0.05). CONCLUSIONS: Diabetes induced morphometric and biomechanical remodelling in intestine. Gliclazide partly restored these changes.

Regulation of von willebrand factor of human endothelial cells exposed to laminar flows: an in vitro study.

The effect of laminar flow on the regulation of von Willebrand Factor (vWF) of cultured human umbilical vein endothelial cells (HUVECs) was studied. Confluent endothelial monolayers were exposed to shear stresses (0.2 and 1.0 Pa) from 2 to 24 h. vWF was labelled with indirect immunofluorescence method and observed with 3D fluorescence microscopy. The distribution of vWF and the cytoskeleton organization were observed simultaneously by double fluorescence labelling. More actin stress fibers and an increased release of vWF appeared in the cells exposed to flow at the same time. The qualitative and quantitative results showed that there was not only a shear-dependent regulation but also a time-dependent modification. For a short-time shear stimulation, both 0.2 Pa and 1.0 Pa shear stresses induced a release of vWF from the endothelial cells. In contrast, after 24 h exposure to 1.0 Pa shear flow, vWFs were much more in the cells than that in the cells exposed to 0.2 Pa for 24 h (p < 0.01) or that in the control cells (p < 0.05). TNF-alpha caused a decrease of vWF and Weibel-Palade bodies in the cells.

Microvascular vasodilatory responses to electric acupuncture in rat brain under acute hemorrhagic hypotension.

The present experiment attempted to evaluate the effect of electrical acupuncture on the cerebral microcirculation in anesthetized rats, using fluorescence videomicroscopy. Changes in the pial arteriolar diameter under acute hemorrhagic hypotension were examined quantitatively. The present results suggest that acupuncture may be effective in improving the cerebral microcirculation in hemorrhagic hypotension.

Effects of ligustrazine on hepatic oxygenation in isolated perfused rat liver.

The effects of ligustrazine on hepatic oxygenation in the isolated rat liver were investigated during prolonged perfusion and following the injection of norepinephrine. After injection of erythrocytes into the perfusate, the hemoglobin spectra in the liver were measured by Erlangen microlightguide spectroscopy, and the hemoglobin oxygenation (HbO2) in the liver was calculated on the basis of the Kubelka-Munk theory. During artificial perfusion, the HbO2 value was decreased from 59.3 +/- 6.4% (after one hour's perfusion) to 25.5 +/- 19.5% (n = 441; after six hours' perfusion). However, when ligustrazine was injected into the perfusate after six hours' perfusion, the HbO2 values recovered to 56.4 +/- 9.7% (n = 441). After injection of norepinephrine, HbO2 in the liver decreased from 48.8 +/- 10.4% to 25.2 +/- 18.4% (n = 961), while subsequent administration of ligustrazine caused a recovery to 62.9 +/- 6.0% (n = 961). Our results suggested that ligustrazine is a powerful hepatic vasodilator for improving hepatic oxygenation.

Measurement of erythrocyte deformability by two laser diffraction methods.

The aim of this work is to study the deformability of red blood cells (RBC) by two laser diffraction methods: the Laser-assisted Optical Rotational Cell Analyser (LORCA, Mechatronics, Amsterdam, Netherlands) and a Shear Stress Diffractometer (RHEODYN SSD, Myrenne, Roetgen, Germany). Experiments were carried out on 46 healthy human subjects. The elongation index EI of normal and hardened RBCs (obtained by heating blood at 49 degrees C or by incubating RBCs in solutions of diamide) was measured. The results showed that the standard deviations of the experimental data for normal RBCs were relatively small, especially at high shear stresses (more than 3.0 Pa), but higher than those reported before. Some correlations between the results given by the two instruments were also found. It should be noted that for hardened RBCs, the standard deviations of the measurements were important compared with the mean values in the two instruments.

In situ expression of ICAM-1 and its mRNA in the lung tissue of asthmatic rats.

In situ expression of ICAM-1 and ICAM-1 mRNA on the lung tissue of asthmatic rats was studied by immunohistochemical staining and in situ hybridization, respectively. The results showed that in normal rats ICAM-1 expression was rare on the endothelium of pulmonary artery and vein, and on the bronchial and alveolar epithelium. The distribution of ICAM-1 expression on the different part of lung tissue of asthmatic rats was similar to that of normal rats, but the level of ICAM-1 expression was significantly increased on the endothelium of pulmonary artery (EPA) and vein (EPV), bronchial epithelium (BEP) and alveolar epithelium (AEP) compared with those of normal and sensitized controls. The distribution and expression of ICAM-1 mRNA on the different part of lung tissue of normal and asthmatic rats were similar to that of ICAM-1 expression. In asthmatic rats, the expression of ICAM-1 mRNA on AEP and EPV was increased significantly compared with those of normal and sensitized controls. It is concluded that the increase of ICAM-1 expression on endothelium of pulmonary vessels, epithelium of broncheoli and alveoli may play an important role of inflammatory cell infiltration in asthmatic rats, and the increased expression of ICAM-1 in asthmatic rats was caused by the increase of expression of ICAM-1 mRNA.

[Effect of Allitridi on deforming of peripheral leucocytes in patients with acute cerebral infarction].

To prove the effect of Allitridi in treating acute cerebral infarction, millipore membrane filter technique was adopted in systematically observing the blood rheology and the peripheral WBC filter index of 53 healthy subjects and that of 33 patients before and after treatments, who suffered from acute cerebral infarction and were confirmatorily diagnosed through CT. As a result, the filtered index of peripheral WBC during the acute period of cerebral infarction rose significantly (6.1397 +/- 4.4602), and the difference was significant compared with that of the healthy subjects (0.8651 +/- 0.4603, P < 0.01). Having been treated with Allitridi, the patients' symptoms improved and at the same time the filtered index of WBC lowered markedly (1.6261 +/- 1.3472). The conditions of the patients before and after treatments were obviously different (P < 0.01). And also the index of the whole blood rheology improved significantly. Therefore, it denoted that Allitridi could effectively improve the WBC deformation and the whole blood rheology as well, and alleviate the clinical symptoms.

Analysis of thromboelastogram on coagulation and fibrinolysis.

Hartert's thromboelastography has been used in the diagnosis of abnormal blood clotting for more than 20 years. From a thromboelastogram three parameters are obtained, viz, the reaction time 'r', the rate of formation of fibrin clot 'k', the maximum elasticity of thrombus 'amax'. It is desirable, however, to know the equation that describes the thromboelastogram both in the period in which the complex modulus increases with time because of coagulation, and in the period in which the complex modulus decreases with time because of fibrinolysis. The parameters of the equation could then be used as a diagnostic criterion; yielding information on the mechanism of coagulation and fibrinolysis. Based on our experimental results on human blood in normal and abnormal subjects, we found that the complex modulus of thromboelastograms can be expressed by the sum of two terms, one describing the increase of the complex modulus during coagulation, G1 = G'1 Exp (-tau 1/t), the other describing the decrease of the complex modulus during fibrinolysis, G2 = G'2 Exp (-tau 2/(t-D) when t greater than D. G2 = 0 when t less than D. The compound complex modulus from coagulation to fibrinolysis is G = G1 - G2. Here t is the clotting time, and G'1, G'2, tau 1, tau 2, and D are five constants to be identified. These five constants can be used for diagnostic and prognostic purposes.

An analysis of the sluicing gate in pulmonary blood flow.

For pulmonary blood flow in zone 2 condition, in which the blood pressure in the venule (pven) is lower than the alveolar gas pressure (pA), the blood exiting from the capillary sheet and entering a venule must go through a sluicing gate. The sluicing gate exists because the venule remains patent while the capillaries will collapse when the static pressure of blood falls below the alveolar gas pressure. In the original theory of sheet flow the effect of the tension in the interalveolar septa on the flow through the sluicing gate was ignored. Since the tension multiplied by the curvature of the membrane is equivalent to a lateral pressure tending to open the gate, and since the curvature of the capillary wall is high in the gate region, this effect may be important. The present analysis improves the original theory and demonstrates that the effect of membrane tension is to cause flow to increase when the venous pressure continues to decrease. The shape of the sluicing gate resembles that of a venturi tube, and can be determined by an iterative integration of the differential equations. The result forms an important link in the theory of pulmonary blood flow in zone 2 condition.

How many pulmonary alveoli are supplied by a single arteriole and drained by a single venule?

A detailed measurement of histological specimens of the lungs of the cat shows that each terminal precapillary vessel (arteriole) supplies, on the average, 24.5 pulmonary alveoli; each terminal postcapillary vessel (venule) drains, on the average, 17.8 alveoli. These numbers link pulmonary alveolar blood flow in capillary sheets with the flow in pulmonary arteries and veins which are cylindrical tubes. They are key numbers needed for hemodynamic analysis. In the literature, these numbers are variously speculated to be 1 or smaller; thus our results correct, even though only for the cat, an important concept.

Morphometry of cat's pulmonary arterial tree.

Morphometic data of the pulmonary artery in the cat's right lung are presented. Silicone elastomer casts of cat's right lung were made, and measured, counted and analyzed. The Strahler system is used to describe the branching pattern of the arterial vascular tree. These data are needed for any quantitative approach to the study of the pulmonary circulation. For all the pulmonary blood vessels of the cat lying between the main pulmonary artery and the capillary beds, there are a total of 10 orders of vessels in the right upper lobe, 9 orders of vessels in the right middle lobe and 11 orders of vessels in the right lower lobe. The ratio of the number of branches in successive orders of vessels or the branching ratio, is 3.58. The corresponding average diameter ratio is 1.72, whereas the average length ratio is 1.81.

Analysis of blood flow in cat's lung with detailed anatomical and elasticity data.

Recently, a complete set of data on the branching pattern of the cat's pulmonary arterial and venous trees and the elasticity of these blood vessels was obtained in our laboratory. Hence it becomes possible for the first time to perform a theoretical analysis of the blood flow in the lung of an animal based on a set of actual data on anatomy and elasticity. This paper presents an analysis of steady flow of blood in cat's lung. The effect of the vessel elasticity is embodied in the "fifth-power law" and the "sheet-flow" theory. The theory yields the pressure-flow relationship of the whole lung, the longitudinal pressure distribution, and the transit time of blood in the capillaries. These results are compared with available experimental data in the literature.

Morphometry of cat pulmonary venous tree.

Morphometric data of the pulmonary veins in the cat right lung are presented. Silicone elastomer casts of the right lungs of five cats were made, measured, counted, and analyzed. The Strahler system is used to describe the branching pattern of the vascular tree. These data are needed for the physicomathematical approach to pulmonary circulation. For all the pulmonary blood vessels lying between the left atrium and the capillary beds, there are a total of 10 orders of vessels in the right upper lobe, 9 orders of vessels in the right middle lobe, and 11 orders of vessels in the right lower lobe. The ratios of the diameters, lengths, and the number of branches in successive orders of vessels are called the diameter, length, and branching ratios, respectively. For the cat pulmonary venous tree, the average branching ratio is 3.521, the average diameter ratio is 1.727, and the average length ratio is 2.402 for vessels of orders 1-3 and 1.532 for vessels of orders 4-10.