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Substrate concentrations generally fluctuate in wastewaters. However, how anammox biomass behaves to overcome the stress of alternating feed loadings remains unclear. Here, we combined long-term reactor operation, batch tests, 16S rRNA transcript sequencing, and metabolomics analysis to investigate the aggregation of anammox biomass under the regulation of c-di-GMP, a key second messenger, in response to alternating feed loadings. We demonstrated that the aggregation process was significantly faster under alternating loadings and was significantly correlated with higher levels of c-di-GMP and extracellular polymeric substances (EPS) production. The increase in c-di-GMP was positively correlated with a higher relative transcript expression level in the c-di-GMP pathway-dependent community. The targeted metabolomics results indicated that the increased production of fructose 6-phosphate and UDP-N-acetyl-d-glucosamine, the precursor substances for the synthesis of exopolysaccharides, was induced by higher levels of c-di-GMP. Consequently, the granulation process was accelerated via EPS production. Higher levels of intracellular hydrophobic amino acids were also positively correlated with increased extracellular protein levels, considering the significant increase in peptides under alternating loadings. On the basis of our findings, we believe that c-di-GMP regulation and EPS production of the anammox biomass are important mechanisms to enhance its tolerance against unfavorable feed stress. These results highlight the role of c-di-GMP in anammox biomass as it works to survive in unfavorable niches.

Multi-class classification algorithm of support vector machine (SVM) has always been a research hotspot. A new multi-class SVM algorithm, naming recall reordering adaptive directed acyclic graphs (RRADAG), is proposed from the perspective of error detection to solve the error accumulation existed in multi-class SVM algorithm of which is based on Directed Acyclic Graphs (DAG). By detecting the output of the classifier and recalling the possible error decision, RADAR has solved the problem of error accumulation. The experimental results generated on UCI standard data sets show that RRADAG outperforms the previously existing algorithms on accuracy.

In this paper, a novel lost packets recovery scheme which encrypts the effective parts of an original packet into two shadow packets based on (2, 2)-threshold XOR-based visual Secret Sharing (VSS) is proposed. The two shadow packets used as watermarks would be embedded into two normal data packets with digital watermarking embedding technology and then sent from one sensor node to another. Each shadow packet would reveal no information of the original packet, which can improve the security of original packet delivery greatly. The two shadow packets which can be extracted from the received two normal data packets delivered from a sensor node can recover the original packet lossless based on XOR-based VSS. The Performance analysis present that the proposed scheme provides essential services as long as possible in the presence of selective forwarding attack. The proposed scheme would not increase the amount of additional traffic, namely, lower energy consumption, which is suitable for Wireless Sensor Network...

The sea-crossing bridge construction takes orders the design idea of maximization, the factorization, the standardization and the assembly, which pile caps and pier shafts of the non-navigable span are prefabricated at the focus of prefabricated. In the paper, the precast solutions are put forward to pile caps and pier shafts of the non-navigable span, and the key technical problems of all kinds of precast solutions are analyzed. On this basis, the optimal pier factory prefabrication is proposed by the comparison of various schemes, which is carried out about advantages and disadvantages of prefabricated matching accuracy control, factory prefabrication, concrete placing, transport in the precast yard, transport style and the impact on follow-up construction.

The cracking and deformation analysis of concrete tunnel segment in the jacking process is made by ANSYS. First, we analyzed the effect of such measures as placing a hydraulic jack between the section and rail beam sliding plane described in the four sliding rails scheme upon the cracking and deformation of segment concrete, then the influence of change in spacing of two sliding rails at the same side on the cracking and deformation of segment concrete stated in the six sliding rails scheme. Meanwhile, we made a comparison between the two layout schemes, thereby providing a basis and reference for the layout scheme of sliding rails in the practice.

Caisson is a commonly used foundation in modern bridge engineering. When the caisson is sinking towards the seabed, it will suffer from the coupled effects of wind, current and its mooring system, under which an intense horizontal oscillation might occur. The mechanism of this kind of oscillation can be concluded as Vortex Induced Motions (VIM) of a large floating object in resilience mooring system. In this paper, through numerical simulation of the viscous flow field around a certain caisson, the mechanism of vortex shedding under different current velocity is discussed.

In the paper, relying on the full-scale model test of embedded pile cap of the HZM Bridge, the method of combining the theoretical calculation analysis and in-situ monitoring test are used to study the slope stability and over-break depth of foundation excavation. In the theoretical calculation analysis, an empirical formula of the Hydtology of Sea-port is adopted for the over-break depth while the G-slope software is for the safety factor of slope stability; in the in-situ monitoring test, the single-beam is used for the topographic survey underwater at random after the foundation excavation. Analysising on the theoretical calculation analysis and the result of in-situ monitoring test, proper slope ratio and over-break depth of pile cap foundation excavation is put forward to provide key construction parameters for the main project of the HZM Bridge.

Antioxidant action is critical for maintaining the normal cardiovascular function and vascular endothelial cell is an important target of estrogen action through estrogen receptor (ER) pathway. This study is carried out to explore the antioxidant effect of carnosol in bovine aortic endothelial cells (BAECs) via ER pathway. The ER subtype specific estrogenic effect of carnosol was further demonstrated by luciferase reporter gene assay in human embryonic kidney (HEK) 293 cells. Carnosol was extracted from Chinese medicine Rosmarinus officinalis. ER positive BAECs were employed in cell proliferation assay and cell apoptosis tests. Oxidative stress by intracellular reactive oxygen species (ROS) were measured via 2'7'-dichlorofluorescein (DCF) production. ERα and ERβ specific antagonists 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole (MPP) and 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidine-3-yl]phenol (PHTPP) were employed as tools in the experiment. ER negative HEK 293 cells were employed in luciferase reporter gene assay. The results indicate that carnosol can effectively attenuate H(2)O(2) induced slowing down of cell growth and increasing of cell apoptosis. At the meantime, carnosol pretreating can also effectively reduce the H(2)O(2) induced intracellular ROS elevation in BAECs. ERα and ERβ antagonist, especially ERα antagonist, can effectively decrease the above antioxidant effects of carnosol. The reporter gene analysis further demonstrates that the action of carnosol on inducing ERE dependent luciferase expression is realized via ER pathway. The conclusion is that carnosol can exert antioxidant effects towards oxidative stress induced by H(2)O(2) in BAECs. And such effects are realized via ER, especially ERα pathway. The results contribute to explain the mechanism of cardiovascular protective function of carnosol in postmenopausal women.

Marinca et al. [1] made some comments on our paper [2] and pointed out “some fundamental mistakes and misinterpretations along with a false conclusion”. Unfortunately, Marinca’s comments are wrong. Here, we further reveal the essence of Marinca’s approach, and point out the reason why their method is indeed time-consuming: their method is nothing more than a traditional method in approximation theory. Numerical results for a given example and related MATHEMATICA code are given to support our view-points.

In this paper, a one-step optimal approach is proposed to improve the computational efficiency of the homotopy analysis method (HAM) for nonlinear problems. A generalized homotopy equation is first expressed by means of a unknown embedding function in Taylor series, whose coefficient is then determined one by one by minimizing the square residual error of the governing equation. Since at each order of approximation, only one algebraic equation with one unknown variable is solved, the computational efficiency is significantly improved, especially for high-order approximations. Some examples are used to illustrate the validity of this one-step optimal approach, which indicate that convergent series solution can be obtained by the optimal homotopy analysis method with much less CPU time. Using this one-step optimal approach, the homotopy analysis method might be applied to solve rather complicated differential equations with strong nonlinearity.

A new triterpenoid saponin, tenuifoside A, was isolated together with three known triterpenoid saponins 2, 3, and 4 from the roots of Polygala tenuifolia Willd. With the help of chemical and spectral analyses (IR, MS, 1D-NMR, and 2D-NMR), the structure of the new saponin was elucidated as 3-O-beta-d-glucopyranosyl presenegenin 28-O-beta-d-xylopyranosyl-(1 --3)-beta-d-xylopyranosyl-(1 --4)-[beta-D-apiofuranosyl-(1 --3)]-alpha-L-rhamnopyranosyl-(1 --2)-[4-O-p-methoxycinnamoyl]-[alpha-l-rhamnopyranosyl-(1 --3)]-beta-d-fucopyranosyl ester (1). Three known triterpenoid saponins (2-4) were identified on the basis of spectroscopic data.

Two new triterpenoid saponins, named cernuaside C and D, have been isolated from Pulsatilla cernua (Thunb.) Bercht. et Opiz. The structures of the two new triterpenoid saponins were elucidated as 3-O-beta-D-xylopyranosyl(1 --2)-[alpha-L-rhamnopyranosyl(1 --3)]-alpha-L-arabinopyranosyl oleanolic acid 28-alpha-L-rhamnopyranosyl(1 --4)-beta-D-glucopyranosyl(1 --6)-beta-D-glucopyranoside. (1) and 3-O-alpha-L-arabinopyranosyl (1 --3)-alpha-L-rhamnopyranosyl(1 --2)-alpha-L-arabinopyranosyl oleanolic acid 28-O-beta-D-glucopyranoside (2) by 1D, 2D-NMR techniques, ESI-MS analysis as well as chemical methods.

Objective The purpose of this research was mainly two-fold: first, to get an understanding of current researches conducted on Alzheimer disease in China; second, to systematically evaluate and compare Alzheimer's treatment delivered by traditional Chinese medicine (TCM) and Western medicine. Methods Two steps were employed in this research. They were data collection and cleaning, followed by systemic review and qualitative analysis. The data were selected from the following two databases: CNKI (http://www.cnki.net) and Wanfang Data (http://www.wanfangdata.com.cn). Inclusion criteria were: (1) Chinese literature; (2) Published between year 1994 and year 2004; (3) Using TCM as treatment and Western medicine as control; (4) Similar research purposes and methodology; (5) Subjects were diagnosed as Alzheimer disease. Descriptive analysis, homogeneity test, meta analysis, sensitivity analysis and subgroup analysis were performed in the second step. Results Supposing all qualified studies were of high quality, we got the following conclusion: the advantage of TCM was losing because of the newly-developed acetylcholinesterase inhibitors came in market. Moreover, the studies conducted after year 2002 were more homogeneous in comparison with those conducted in early years. Those studies using mini-mental status examination (MMSE) as outcome measurement were also more homogeneous than non-MMSE measurement groups. Combined odds ratio in comparative studies was 1.5 fold higher than that in experimental studies. Regarding to different outcome measurement, those studies using TCM assessment profile were 2.58(4.79/1.86) fold higher than those using MMSE as outcome measurement. Conclusions After systemic literature search, we found that only 40 out of 2,403 studies met our inclusion criteria. Moreover, those qualified studies were of low quality. Therefore, the external validity of this study would be compromised. The solution to this would be to improve study quality by strengthening study design and carefully select more homogeneous subjects in terms of syndrome differentiation, by so doing, the results of meta analysis will be more convincing and easily recognized by international society.

In order to obtain optimum decision making in selecting main diesel engine for ships, the life cycle cost technology is proposed. The breakdown structure and calculation model of life cycle cost for main diesel engine of ships are established. Taking a specific selection for main diesel engine of one ship as the instance, the purchase cost and life cycle cost for three types of main diesel engine are compared and analyzed. The type of main diesel engine is selected based on the calculation result of life cycle cost. The instance shows that the life cycle cost technology is a powerful decision-making tool, which is worthy of popularization and application in ship design and development stages.

Carnosol has been proved to have anti-breast cancer effect in previous research. But its ER subtype's specific regulation and mediation mechanisms remain unclear. The aim of this study is to observe the effect of carnosol on cell proliferation and its estrogen receptor α and β's specific regulation and mediation mechanisms with ER positive breast cancer T47D cell. With estrogen receptor α and β antagonists MPP and PHTPP as tools, the MTT cell proliferation assay was performed to observe the effect of carnosol on T47D cell proliferation. The changes in the T47D cell proliferation cycle were detected by flow cytometry. The effect of carnosol on ERα and ERβ expressions of T47D cells was measured by Western blot. The findings showed that 1 x 10(-5)-1 x 10(-7) mol x L(-1) carnosol could significantly inhibit the T47D cell proliferation, which could be enhanced by MPP or weakened by PHTPP. Meanwhile, 1 x 10(-5) mol x L(-1) or 1 x 10(-6) mol x L(-1) carnosol could significantly increase ERα and ERβ expressions of T47D cells, and remarkably increase ERα/ERβ ratio. The results showed that carnosol showed the inhibitory effect on the proliferation of ER positive breast cancer cells through target cell ER, especially ERβ pathway. In the meantime, carnosol could regulate expressions and proportions of target cell ER subtype ERα and ERβ.

Dynamic responses of a heavy cycloidal object hanged by a large floating crane in the process of immersing into waves were studied experimentally and numerically in this paper. A floating crane with displacement of 14000t was selected as the reference. A model with the scale of 1:20 was conducted in a towing tank. The movement responses and forces of the heavy cycloidal object in the process of immersing in waves were researched. Ocean engineering construction is always affected significantly by waves, so the selection of a shield to attenuate incoming waves is an important aspect for ocean engineering construction. The sea condition in our project is 3-state sea condition; it’s not an acceptable condition for real construction considering safety of crews and the ability of equipments. So an effective floating breakwater validated experimentally in a towing tank in our project was proposed as effective shields. The steel cylinder lifted by a floating crane in the process of water-entering was experimented, the movements of the steel cylinder and the subjected forces of the hoist were recorded; the effects of different shields on the responses of steel cylinder with different depth in water were studied. Numerical simulations of the motion of steel cylinder subjected to waves in the process of immersing were conducted to illustrate the effects of headings. The basis of model design, model-making and measurement methods are also introduced in this paper. The results are translated into full scale results which will be the basis for safety construction of ocean engineering and measurement proposed for accurate positioning and installation of the steel cylinder.

As the development of ocean engineering, safety construction is becoming a significant aspect that attracted more and more attention. The general aims of this paper were to present the effects of different floating breakwater on wave absorbing and provide a basis for the design of floating breakwater of an offshore project. Three different kinds of floating breakwater were studied; they are soft wall-sided pontoon, solid wall-sided pontoon and a circular cross-section breakwater filled with water. Various factors including wave period, wave steepness and geometrical characteristics were considered. The results had shown that the three proposed floating breakwater revealed favorable performance and the geometrical configuration could impact wave attenuation significantly. The circular cross-section breakwater had greater effects on wave absorbing compared with two other floating breakwaters, more detailed experiments were conducted to the circular cross-section breakwater, a V-shaped floating breakwater consists of two single circular cross-section breakwater were studied, the effects of angle of the two singles, wave period and wave height were considered. The results revealed that the geometrical configuration and water depth could impact wave attenuation and mooring force significantly. Researches are expected to offer some information for the design of floating breakwater for an offshore project.Copyright © 2014 by ASME

Antioxidant action is critical for maintaining the normal cardiovascular function and vascular endothelial cell is an important target of estrogen action through estrogen receptor (ER) pathway. This study is carried out to explore the antioxidant effect of carnosol in bovine aortic endothelial cells (BAECs) via ER pathway. The ER subtype specific estrogenic effect of carnosol was further demonstrated by luciferase reporter gene assay in human embryonic kidney (HEK) 293 cells. Carnosol was extracted from Chinese medicine Rosmarinus officinalis. ER positive BAECs were employed in cell proliferation assay and cell apoptosis tests. Oxidative stress by intracellular reactive oxygen species (ROS) were measured via 2'7'-dichlorofluorescein (DCF) production. ERα and ERβ specific antagonists 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole (MPP) and 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidine-3-yl]phenol (PHTPP) were employed as tools in the experiment. ER negative HEK 293 cells were employed in luciferase reporter gene assay. The results indicate that carnosol can effectively attenuate H(2)O(2) induced slowing down of cell growth and increasing of cell apoptosis. At the meantime, carnosol pretreating can also effectively reduce the H(2)O(2) induced intracellular ROS elevation in BAECs. ERα and ERβ antagonist, especially ERα antagonist, can effectively decrease the above antioxidant effects of carnosol. The reporter gene analysis further demonstrates that the action of carnosol on inducing ERE dependent luciferase expression is realized via ER pathway. The conclusion is that carnosol can exert antioxidant effects towards oxidative stress induced by H(2)O(2) in BAECs. And such effects are realized via ER, especially ERα pathway. The results contribute to explain the mechanism of cardiovascular protective function of carnosol in postmenopausal women.

To study the protective effect of soybean protease inhibitor on LPS-induced lung injury in rats.Fifty male SD rats were randomly divided in five groups, 10 rats in each group as sham-operation group, model control group, positive medicine group, and high, moderate SBTI groups. Except the sham-group, other groups were induced by intratracheal instillation of LPS with a dose of 6 mg x kg(-1). All rats were given drug throughout intraperitoneal injection except the model controlled group, the positive medicine group was given PMSF with a dose of 50 mg x kg(-1), the high dose group of SBTI was given SBTI with a dose of 100 mg x kg(-1), a dose of the moderate group is 50 mg x kg(-1). We examined all rats in seven days. Index exam: cell quantity, activity of neutrophilic granulocyte released elastic protease proteins in BALF, histopathological examination and so on.Soybean protease inhibitor can level down the level of total protein, cell quantity, PMN percent, activity of neutrophilic granulocyte in BALF. SBTI level down the content of NF-kappa B in nucleoprotein, while increase the content of I kappa B alpha in plasmoprotein.SBTI is useful in protecting experimental pulmonary injury induced by LPS in rats.

A new triterpenoid, saponin hehuanoside A, was isolated together with the known triterpenoid saponins 2, 3, and 4 from the stem bark of Albizia julibrissin. With the help of chemical and spectral analyzes (IR, MS, 1D-NMR, and 2D-NMR), the structure of the new triterpenoid saponin was elucidated as 21-O-[(6S)-2-trans-2,6-dimethyl-6-O-beta-d-quinovopyranosyl-2,7-octadienoyl]-3-O-beta-d-xylopyranosyl-(1 --2)-beta-d-fucopyranosyl-(1 --6)-beta-d-2-deoxy-2-acetamidoglucopyrasyl acacic acid 28-O-alpha-l-arabinofuranosyl-(1 --4)-[-beta-d-glucopyranosyl-(1 --3)]-alpha-l-rhamnopyranosyl-(1 --2)-beta-d-glucopyranosyl ester (1). Three known triterpenoid saponins 2-4 were identified on the basis of spectroscopic data.

To investigate the effects of bleomycin (BLM) on the apoptosis of type II alveolar epithelial cell (AT II) in lung fibrotic rats and its possible mechanisms.Totally 32 male Sprague-Dawley rats were randomly divided into sham group (n = 8) and BLM group (n = 24). Rats in sham group or BLM group were intratracheally instillated with saline or 5 mg/kg of bleomycin, respectively. One, three, and seven days after the instillation of bleomycin, 8 rats in BLM group were taken for AT II isolation and purification. Rats in sham group were used to isolate and purify AT II on 7 days after the instillation of saline. The cell cycle and apoptosis, intracellular free calcium concentration, and mitochondrial membrane potential (MMP) in AT II were determined by flow cytometry. Immunohistochemistry was performed to observe the expressions of Bax, Bcl-2, and Fas. Caspase-3, Caspase-8, and Caspase-9 activities were measured by Caspase activity detection kit.The ratio of S phase AT II in BLM group was significantly lower than in sham group (P0.05). AT II apoptosis rates on day 1 and 3 were significantly higher in BLM group than in sham group (P0.01). Intracellular free calcium concentrations in BLM group were significantly higher than in sham group (P0.05). However, MMP was significantly lower than sham group (P0.05). The positive rates of Bax, Fas and Caspase-3, Caspase-8, and Caspase-9 activities of BLM group were significantly higher than those of sham group (P0.05, P0.01). The positive rates of Bcl-2 on day 1 and 3 were significantly lower than those of sham group (P0.05).Early AT II apoptosis may be induced by bleomycin, which may be explained by the increase of intracellular free calcium concentration, depression of MMP, increased expressions of Fas and Bax, and increase of Caspase-3, Caspase-8, and Caspase-9 activities.

A new furospirostanol saponin, ophiofurospiside A (1), was isolated together with the known steroidal glycosides 2, 3, and 4 from the tubers of Ophiopogon japonicus (Thunb.) Ker-Gawl. Using chemical and spectral analyses (IR, MS, 1D NMR, and 2D NMR), the structure of 1 was established as 26-O-beta-d-glucopyranosyl-(22S, 25R)-furospirost-5-ene-3beta, 17alpha, 26-triol-3-O-[alpha-l-rhamnopyranosyl-(1 --> 2)]-[beta-d-xylopyranosyl-(1 --> 4)]-glucopyranoside (1). Three known steroidal saponins 2-4 were identified on the basis of spectroscopic data.

Two new furostanol saponins, tribufurosides B (1) and C (2), were isolated from the fruits of Tribulus terrestris L. With the help of chemical and spectral analyses (IR, MS, 1D NMR and 2D NMR), the structures of two new furostanol saponins were established as 26-O-beta-d-glucopyranosyl-(25S)-5alpha-furost-20(22)-en-2alpha,3beta,26-triol-3-O-beta-d-galactopyranosyl(1 --2)-beta-d-glucopyranosyl(1 --2)-beta-d-galactopyranoside (1) and (25S)-5alpha-furost-20(22)-en-12-one-3beta, 26-diol-26-O-beta-d-glucopyranoside (2).

To observe and evaluate the phytoestrogenic effects and its mechanism of psoralen in estrogen receptor (ER) alpha and beta positive T47D and ishikawa cells.The proliferation rate of T47D influenced by 1 x 10(-5) mol x L(-1) to 1 x 10(-9) mol x L(-1) psoralen and that of Ishikawa influenced by 1 x 10(-6) mol x L(-1) and 1 x 10(-7) mol x L(-1) psoralen were analyzed by MTT assay. PR mRNA expression in T47D was quantified by RT-PCR assay. Estrogen receptor antagonist ICI 182, 780 was employed as a tool. ER-alpha and ER-beta expression of T47D was measured by flow cytometry.The proliferation rates of T47D cells treated with 1 x 10(-5) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen and ishikawa cells treated with 1 x 10(-6) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen were increased significantly. The RT-PCR result showed that 1 x 10(-7) mol x L(-1) and 1 x 10(-6) mol x L(-1) psoralen could increase PR expression in T47D cells. The above effects could be blocked by ICI 182,780. Psoralen could also induce the augment of ER-alpha and ER-beta expression in T47D cells significantly.Psoralen has phytoestrogenic effects. The effects are attained through ER pathway.

Gamma-glutamyl transpeptidase (γ-GTP) was localized in prenatal and neonatal rat kidneys using an immunohistochemical method with monoclonal antibodies to rat kidney γ-GTP. γ-GTP was demonstrated on the brush border and basolateral membrane of proximal convoluted tubules from the 18th fetal day. The basolateral membrane of proximal tubules revealed strong reaction at 1 and 3 days after birth. The reaction of basolateral membrane decreased with the maturation of nephrons, however, moderate reaction was still observed at 30 days of age in the S3 segment. Throughout the postnatal development of the kidney, the S3 exhibited the strongest immunostainingand S1 showed the weakest reaction, and proximal tubules in the deep layer of the cortex always provided a stronger reaction than those in the superficial layer of the cortex. Maturating cuboidal and squamous epithelial cells of descending Henle's loop showed a positive reaction on the luminal and lateral cell membranes. On immunoelectron microscopy, γ-GTP was found on the brush border, basolateral membrane, membranes of pinocytotic vesicles and apical invaginations in proximal tubular cells at 1-7 days after birth. However, tight junctions were devoid of the reaction. The development of basal infolding and localization of γ-GTP there seemed to relate closely to the development of surrounding capillaries. γ-GTP was localized on the cell membrane and occasionally in the cytoplasm of interstitial cells of renal interstitium.

To explore the phytoestrogenic effects of ten kinds of Chinese medicine including flos carthami, radix cyathulae, radix salviae miltiorrhizae, fructus ligustri lucidi, fructus lycii, radix clycyrrhizae, herba cistanches, herba epimedii, fructus psoraleae and semen cuscutae.240 female Kunming mice weighting 9 - 12 g were randomly divided into two main groups A and B. A group was divided into 12 small groups: 1 solvent control group, 1 diethylstilbestrol control group and 10 Chinese medicine groups. B group was also divided into 12 small groups: 1 solvent control group, 1 diethylstilbestrol control group and 10 Chinese medicine antagonistic groups. Mice in ten antagonistic groups were administered both Chinese medicine and diethylstilbestrol everyday. After administered(op) for 4 days, blood was collected and serum was separated. The effect of the pharmacological serum on proliferation rate of MCF-7 (ER+) was analyzed by MTT-assay.In A group, proliferation rates of MCF-7 cells treated with serum from eight Chinese medicine groups including flos carthami, radix cyathulae, radix salviae miltiorrhizae, fructus lycii, herba cistanches, herba epimedii, fructus psoraleae and semen cuscutae were coued markedly increase respectively. While serum from fructus ligustri lucidi group could markedly decrease the proliferation rate of MCF-7 cells. In B group, the increased proliferation rate of MCF-7 cells caused by diethylstilbestrol was significantly reduced in seven Chinese medicine antagonistic groups including flos carthami, radix cyathulae, radix salviae miltiorrhizae, radix clycyrrhizae, herba epimedii, fructus psoraleae and semen cuscutae. While the increased proliferation rate could be markedly enhanced in herba cistanches group.Six kinds of Chinese medicine such as flos carthami, radix cyathulae, radix salviae miltiorrhizae, herba epimedii, fructus psoraleae and semen cuscutae show both estrogenic effects (when administered indepently) and antiestrogenic effects (when administered together with diethylstilbestrol). Such bidirectional effects depends on the internal estrogen level.

To observe the changes of matrix metalloproteinases (MMPs) activities in pulmonary fibrosis rats.Eighty male SD rats were randomly divided into sham group (n = 40) and bleomycin group (BLM, n = 40), in which SD rats were injected with a single intratracheal dose of sham saline or bleomycin respectively. On day 1, 3, 7, 14, and 28 following bleomycin or saline instillation, rats were randomly killed, and serum from abdominal aorta, alveolar fluid from the bronchoalveolar lavage, and the lung homogenate were collected and then stored at -80 degrees C. MMPs activity was determined by zymography.Compared with sham group, the levels of MMP-9 in all samples were augmented. MMP-9 activities in the serum were highest on day 3 than those on day 1 and day 7, and in lung tissue homogenate were highest on day 7; however, no significant differences were found between BLM group and sham group on day 14 and day 28; and that of bronchoalveolar lavage fluid (BALF) was highest on day 7 than those on day 1 and day 14, while no significant difference existed between BLM group and sham group on day 28. Serum MMP-2 level did not change from day 1 to day 28, while the level of BALF MMP-2 began to increase after day 14, even on day 28. Lung tissue homogenate MMP-2 level began to increase early on day 3 and continued to day 28.The sources and effects of MMP-2 and MMP-9 differ in BLM-induced rat pulmonary fibrosis.

To evaluate the estrogenic activity of several kinds of Chinese herbal medicines.Use zoopery and reporter gene technique to study the estrogenic activity of five Chinese herbal medicines. Zoopery: weanling female Kunming mice weight 9-12 g were administrated botanical extracts of Selaginella tamariscina, Pinus Massoniana, Corallodiscus flabellate, Dryopteris sublaeta and Leonurus heterophyllus, the positive control group with Nilestriol tablets and control group with water, respectiely. On the eighth day, the animals were sacrificed and the uteri were separated solely and weighed. Reporter gene technique: Induce the expression of reporter gene controlled by ERE and measure the activity of luciferase on cell's clear supernatant.The botanical extracts of S. tamariscina can increase weights of mice (P0.01); In the expression of reporter gene controlled by ERE, Either ERalpha or ERbeta's has estrogenic activity (P0.01). Follow in the zoopery we find the water part and the n-butanol part of S. tamariscina are the two active parts.S. tamariscina and it's water part and n-butanol part have estrogenic activities, effect on ERbeta is greater than ERalpha.

To study the protective effects of curcumin on exaggerated extracellular matrix accumulation of pulmonary fibrosis rats.One hundred and forty-four male Sprague-Dawley rats were randomly divided into 6 groups (24 rats in each group). Rats in the model control group, positive medicine group, and high, moderate and low curcumin groups were injected with a single dose of bleomycin by trachea, and rats in sham-model control group with same volume normal saline. One day after the injection, curcumin solution of different dosages (200, 100, 50 mg x kg(-1) x d(-1)) was respectively given to rats in the high, moderate and low curcumin group daily by gastrogavage, while equal volume of normal saline was given to those in the sham-model control group and model control group, and an equal volume of prednisone (0.56 mg x kg(-1) x d(-1)) was given to those in positive medicine control group. On the 7, 14, 28 days, 8 rats per treatment group were randomly killed, the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum were determined, the determination of hydroxyproline in lung homogenates was analyzed, and the lung was incised to make pathological sections which were stained with HE and Mallory.Curcumin could decreas the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum, and inhihit the proliferation of fibrous tissue.Curcumin may play its therapetuic role by leveling down the content of extracellular matrix in rats with pulmonary fibrosis induced by bleomycin.

AIM: To study the role of hepatic sinusoidal capillarization and perisinusoidal fibrosis in rats with alcohol-induced portal hypertension and to discuss the pathological mechanisms of alcohol-induced hepatic portal hypertension. METHODS: Fifty SD rats were divided into control group (n=20) and model group (n=30). Alcoholic liver fibrosis rat model was induced by intragastric infusion of a mixture containing alcohol, corn oil and pyrazole (1 000:250:3). Fifteen rats in each group were killed at wk 16. The diameter and pressure of portal vein were measured. Plasma hyaluronic acid (HA), type IV collagen (CoIV) and laminin (LN) were determined by radioimmunoassay. Liver tissue was fixed in formalin (10%) and 6-μm thick sections were routinely stained with Mallory and Sirius Red. Liver tissue was treated with rabbit polyclonal antibody against LN and ColIV. Hepatic non-parenchymal cells were isolated, total protein was extracted and separated by SDS-PAGE. MMP-2 and TIMP-1 protein expression was estimated by Western blotting. RESULTS: The diameter (2.207 ± 0.096 vs 1.528 ± 0.054 mm, P

To explore the possible effects and mechanism of Fufang Biejiafang on a single intratracheal instillation (IT) of bleomycin-induced lung fibrosis model.SD rats were treated with a single IT dose of bleomycin or control saline. Chinese medicine group were poured into the stomach after the first day of operation with high dosage, middle dosage and low dosage. On days 7, 14 and 28 following IT bleomycin or saline, 4 mL blood were taken from the abdominal aorta for arterial blood gas analysis. The left lung was fixed for routine light microscopic examination. Bronchoalveolar lavage fluid (BALF) from the right lung was tested the activity of pulmonary surfactant (PS) by the Whihelmy Film Balance, then the right lung was frozen immediately in liquid nitrogen for determination of hydroxyproline concentration.Model rats had obviously changes of body weight and hypoxemia and dysfunction of PS on days 7 and improved on days 14. Compared with three dose groups, the middle dose group some degreely improved and PS function. It ameliorate fibrosis because of inhibition of inflammation.(1) PS dysfunction resulted in hypoxemia after bleomycin injured alveolar type II (AT II). Fufang biejiafang-middle dose-group ameliorate hypoxemia by remission AT-II injury. (2) Fufang biejiafang may inhibit exudation inflammation and ameliorate fibrosis.

To study the influence of compound Biejia Ruangan Prescription (CBRP) on extracelluar matrix in bleomycin induced pulmonary fibrosis rats.54 male Sprague-Dawley rats were randomly divided into 6 groups (9 rats in each group). Rats in the model control group, positive medicine group, and high, moderate and low CBRP groups were injected with bleomycin A5 by trachea, and rats in sham-model control group with same volume normal saline. 29 days after the injection, CBRP solution of different dosages (1.4 g x kg(-1), 0.7 g x kg(-1), 0.35 g x kg(-1)) was respectively given to rats in the high, moderate and low CBRP group by gavage, while equal volume of normal saline was given to those in the sham-model control group and model control group, and an equal volume of prednisone (0.56 mg x kg(-1)) was given to those in positive medicine control group. On the 80th day, the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum were determined, the determination of hydroxyproline in lung homogenates was analyzed, and the right lung was incised to make pathological sections which were stained with Hematoxylin-Eosin (HE) and Masson staining for pathological diagnosis.CBRP could decrease the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum.CBRP may play its therapeutic role by leveling down the content of extracellular matrix in rats with pulmonary fibrosis induced by Bleomycin A5.

To study the chemical constituents of Selaginella tamariscina (Beauv.) Spring.The compounds were isolated and purified by macroporous adsorption resin, Sephadex LH-20 and silica gel column chromatography and identified on the basis of their physicochemical and spectral data.Four compounds were obtained from the n-BuOH fraction of 70% acetone extracts. Their structures were elucidated as (7S, 8R)-7, 8-dihydro-7-(4-hydroxy-3,5-dimethoxyphenyl)-8-hydroxymethyl-[1'-( 7'-hydroxyethyl)-5' methoxyl] benzofuran-4-O-beta-D-glucopyranoside (tamariscinoside C, I), D-mannitol (II), tyrosine (II), shikimic acid (IV).Compound I is a new compound, compounds II and III were obtained from the genius for the first time, compound IV was yielded from the plant for the first time.

To study the chemical constituents of Selaginella tamariscina (Beauv.) Spring.Various chromatographic techniques were used to separate and purify the chemical constituents. Their physico-chemical properties and spectral data were used to elucidate the structures.Four compounds were isolated from the n-BuOH fraction of the water-extracts. Their structures were identified as 1-hydroxy-2-[2-hydroxy-3-methoxy-5-(1-hydroxyethyl)-phenyl]-3-(4-hydroxy-3,5-dimethoxy)-propane-1-O-beta-D-glucopyranoside (tamariscinoside B, I), adenosine (II), guanosine (III), arbutin (IV).Tamariscinoside B (I) is a new compound, while the others were isolated from Selaginella for the first time.

AIM To establish a drug screening model based on transcriptional regulation of estrogen responsive element (ERE) and use it to screen compounds for discovering new ligands of estrogen receptor (ER) subtypes.A recombinant reporter vector pERE-TAL-SEAP was constructed by inserting a synthetic sequence composed of five tandem copies of EREs upstream of promoter of the reporter vector pTAL-SEAP. The pERE-TAL-SEAP and the internal control plasmid pCMV were transiently co-transfected into Hela cells expressing ER subtype or ER subtype, and the effects of pure ER agonists 17estradiol, phytoestrogen genistein and pure ER antagonist ICI182, 780 on reporter gene SEAP expression were observed.In the Hela cells expressing ER alpha or ER beta subtype, the expression of SEAP gene were induced in a dose dependent manner by 17-estrodiol with a maximal effect at approximately 10 nmol.L-1 and with EC50 of (80.58 +/- 8.51) pmol.L-1 and (103.90 +/- 5.29) pmol.L-1, respectively, so done by phytoestrogen genistein with a maximal effect at 1 mumol.L-1 and with EC50 of (10.86 +/- 0.75) nmol.L-1 and (39.38 +/- 2.26) nmol.L-1, respectively. The maximal level induced by estrodiol and genistein were about 7-14 fold higher than that of vehicle. The pure antiestrogen ICI182, 780 at concentration of 1 mumol.L-1 completely blocked the inductions of 17-estrodiol and genistein.The cellular drug screening model can be established by transfecting reporter vector pERE-TAL-SEAP in Hela cell lines expressing ER alpha or ER beta. The cell lines can be used to screen compounds with estrogenicity by testing SEAP activity in the culture media of cells growing in microtitier wells. The system should provide an efficient model for screening and analyzing the activity of large numbers of ligands of ER.

To study the effects of extracts of root of kudzu vine on mammary gland and uterus development in rats.40 Wistar rats weighting 65-85 g were randomly divided into 4 groups: control group, estrogen group, extracts of root of kudzu vine group of high dose, extracts of root of kudzu vine group of low dose. (10 rats in each group). After having been treated for 7 days, the rats were killed; mammary glands and uterus were removed and weighed. Serum was isolated and kept at 4 degrees C for determination of hormones.1. Administration of the root of kudzu vine significantly increased the weigh of mammary gland and uterus in rats. 2. Administration of the root of kudzu vine increased serum FSH, LH, E2 and decreased PRL.Extracts of root of kudzu vine could enhance the weight of mammary gland and uterus growth in rats, which may provide experimental evidence for the development of new drug used for promoting mammary gland and uterus.

To study the inhibitig effects of genistein on the growth of human breast cancer cell lines in vitro and its mechanisms. METHORD: Human breast cancer cell lines both MCF-7(positive estrogen receptor, ER+) and MDA-MB-231 (negative estrogen receptor, ER-) were cultured in vitro. The proliferation of cells was measured with MTT methord and the growth curve was drawn with cell count. The estrogen receptor in cells was show with immunohistochemistry.Genistein inhibited proliferation of MCF-7 and MDA-MB-231 cell lines and inhibition was dependent on dose within some concentration range, IC50 being 32.5 mumol.L-1 and 46.8 mumol.L-1 respectively. Genisteins antiproliferation of MCF-7 was stimulated by ectogenesis estrogen but proliferation of MDA-MB-231 inhibited by geinstein was not related to estrogen. The positive signs of ER in cellular nuclei of MCF-7 cell line fed with genistein at concentration of 30 mumol.L-1 were significantly weaker than these not fed with genistein.Genistein obviously inhibits proliferation of both cell lines of MCF-7 and MDA-MB-231 in vitro and is dependent on dose. Genisteins antiproliferous effect on MCF-7 cell lines is stimulated by estrogen and this effect is related with ER, but genisteins inhibiting proliferation of MDA-MB-231 cell line is not through ER.

In order to study on effects of soy extract on energy metabolims in ovariectomized rats.90 Wistar rats were randomly divided into 9 groups: control group, sham group, model group, estrogen group, soy isoflavone group of high dose, soy isoflavone of low dose, soy extract of high dose, soy extract of low dose, 10 rats each group. Beside of control and sham groups, the rest rats were ovariectomized. One week after operation, the rats were treatmented with different drugs, measument of body weigh and feed weigh each week. Six week after operation, the rats were killed, serum were taken, abdomen lipid were removed and weight.The ovariectomized rats took more food and got weight gain significantly; Body mess index(BMI), Abdomen lipid weigh and food transform rate in Model group increased significantly than control and sham groups. Administration of estrogen or soy extract or soy isoflavone could block these changes in ovariectomized rats, but soy polysaccharides did not have the effects.Ovariectomized rats have imbalance of energy metabolism, weigh gain and accumulation of abdomen lipid; administration of estrogen, soy extracts or soy isoflavone could attenuate these changes induced by ovariectomizing.

To study effects of soy extract on lipid metabolims in ovariectomized rats.90 Wistar rats were randomly divided into 9 groups: control group, sham group, model group, estrogen group, soy isoflavone group of high dose, soy isoflavone group of low dose, soy extract of high dose, soy extract of low dose, and soy polysaccharde group, 10 rats in each group. Except fer of control and sham groups, the test rats were ovariectomized. One week after operation, the rats were treated with different drugs. Six weeks after operation, the rats were killed, with serum and liver taken, and serumglycerol(sGT), cholesterol(sGC), LDL, HDL and liver homogenate hGT, hGC, measured.The level of sGC, LDL in ovariectmized rats increased significantly, compared with that in control and sham groups. In liver both the level of hGT and hGC were higher than that in liver from control and sham groups. Administration of estrogen or soy extract or soy isoflavone could attenuate these in ovariectomized rats, but soy polysacchardes did not have any effects.Ovariectomized rats have an imbalance of lipid metabolism, the level of hGT and hGC were increased, and administration of estrogen, soy extracts or soy isoflavone could decrease these changes induced by ovariectomizing.

AIM: To study the effects of Radix Puerariae flavones (RPF) on liver lipid metabolism in ovariectomized (OVX) rats. METHODS: Forty adult female Wistar rats were randomly divided into four groups: OVX group; sham-OVX group; OVX + estrogen group and OVX + RPF group. One week after operation rats of the first two groups were treated with physiological saline, rats of OVX + estrogen group with estrogen (1 mg/kg·b.w.) and rats of OVX + RPF group with RPF (100 mg/kg·b.w.), respectively for 5 weeks. After the rats were killed, their body weight, the weight of the abdominal fat and uterus were measured, and the levels of total cholesterol (TC) and triglyceride (TG) in liver homogenate were determined. RESULTS: Compared with the sham-OVX group, the body mass of the rats in OVX group was found increased significantly; more abdominal fat in store; TC and TG in liver increased and uterine became further atrophy. As a result, the RPF was found to have an inhibitive action on those changes of various degrees. CONCLUSION: RPF has estrogen-like effect on lipid metabolism in liver and adipose tissue.

AIM: To observe the effect of compound Biejiaruangan decoction (CBJRGC) (composite prescription of Carapax trionycis for softening the liver) on proliferation, activation, excretion of collagen and cytokine of hepatic stellate cells (HSCs) and to find the mechanism of prevention and treatment of hepatic fibrosis by CBJRGC. METHODS: Using MTT, immunohistochemistry and image analysis technology, the related indexes for proliferation, activation, excretion of collagen and cytokine of hepatic stellate cells were detected in 24 h, 48 h, and 72 h after adminstration of different dosages of CBJRGC. RESULTS: Statistical analysis showed that serum collected from rat perfused with CBJRGC could restrain the proliferation of HSC in 48 h and 72 h especially in high and medium dosage groups, markedly decrease the expression of desmin, synapsin and platelet derived growth factor (PDGF) in HSC in 24 h, 48 h and 72 h, as well as the expression of α-SMA, collagen III, TIMP and TGFβ1 in 48 h and 72 h, decrease the excretion of collagen I in 72 h. CBJRGC serum had no significant effect on collagens I, III and TIMP in 24 h. CONCLUSION: CBJRGC serum has a good curative effect on hepatic fibrosis. Its main mechanism may be related to the following factors. The drug serum can restrain the proliferation and activation of HSC, decrease the number of activated HSC and the total number of HSC, the excretion of collagens I, III, enhance the degradation of collagen and restore the balance of synthesis and degradation of collagen, inhibit the expression of transforming growth factor β1 (TGFβ1) and platelet derived growth factor (PDGF) in HSC, block and delay the process of hepatic fibrosis. Synapsin is a new marker of activation of HSC, which provides a theoretical and testing basis for neural regulation in the developing process of hepatic fibrosis.