Choline induced cardiac dysfunction by inhibiting the production of endogenous hydrogen sulfide in spontaneously hypertensive rats.

To investigate the exact effects of dietary choline on hypertensive heart disease (HHD) and explore the potential mechanisms, male spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were randomly divided into five groups as follows: WKY group, WKY + Choline group, SHR group, SHR + Choline group, and SHR + Choline + NaHS group. In choline treatment groups, rats were fed with 1.3% (w/v) choline in the drinking water for 3 months. The rats in the SHR + Choline + NaHS group were intraperitoneally injected with NaHS (100 micromol/kg/day, a hydrogen sulfide (H2S) donor) for 3 months. After 3 months, left ventricular ejection fraction (LVEF) and fractional shortening (LVFS), the indicators of cardiac function measured by echocardiography, were increased significantly in SHR as compared to WKY, although there was no significant difference in collagen volumes and Bax/Bcl-2 ratio between the two groups, indicating the early stage of cardiac hypertrophy. There was a significant decrease in LVEF and LVFS and an increase in collagen volumes and Bax/Bcl-2 ratio in SHR fed with choline, meanwhile, plasma H2S levels were significantly decreased significantly in SHR fed with choline accompanying by the decrease of cystathionine-gamma-lyase (CSE) activity. Three months of NaHS significantly increased plasma H2S levels, ameliorated cardiac dysfunction and inhibited cardiac fibrosis and apoptosis in SHR fed with choline. In conclusion, choline aggravated cardiac dysfunction in HHD through inhibiting the production of endogenous H2S, which was reversed by supplementation of exogenous H2S donor.

[Component analysis of Ophiocordyceps lanpingensis polysaccharides and study on alleviation of hepatic fibrosis in mice by polysaccharides].

Ophiocordyceps lanpingensis is mainly used as an ethnic medicine to treat the diseases of liver, kidney and other diseases, but the pharmacological mechanism is not clear yet. In this study, the components and contents of monosaccharides in the O.lanpingensis polysaccharides(OLP) were analyzed. The results showed that the OLP were composed of mannose, glucose, galactose and arabinose, with mass percentages of 19.1%, 21.8%, 21.1%, and 38.0%, respectively. Based on the hepatic fibrosis model induced by CCl_4 in mice, OLP could significantly relieve the inflammation and fibrosis levels of hepatic tissues, reverse the CCl_4-induced increasing levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in mice serum, and recover the functions of liver to a normal state. This study proved that OLP significantly decreased the mRNA expression levels of fibrotic genes, alpha-smooth muscle actin(α-SMA) and collagen type 1(Col-1), as well as the content of hydroxyproline(HYP) in the liver tissues; meanwhile, the contents of antioxidants superoxide dismutase(SOD) and glutathione(GSH) were enhanced and the production of lipid peroxide malondialdehyde(MDA) was reduced. Moreover, OLP inhibited the gene expression levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and nuclear factor kappa B(NF-κB) in the livers of mice. Further study indicated that OLP could restrain the apoptosis of hepatic cells due to the decrease of the apoptosis index and down-regulations of protein expression levels of Bcl-2 associated X protein(Bax), cysteinyl aspartate specific proteinase-3(caspase-3) and cysteinyl aspartate specific proteinase-9(caspase-9), and the promotion of protein expression level of B-cell lymphoma-2(Bcl-2) in livers. To sum up, the mechanism of OLP for alleviating hepatic fibrosis was likely related to the synergy by remitting the oxidative stress of the body, alleviating inflammatory response, anti-apoptosis of hepatic cells, and so on.

Nanoporous anodic alumina reduces Staphylococcus biofilm formation.

Staphylococcus epidermidis and Staphylococcus aureus, two bacterial strains commonly associated with biofilm-related medical infections and food poisoning, can rapidly colonize biotic and abiotic surfaces. The present study investigates the ability of anodic alumina surfaces with nanoporous surface topography to minimize the attachment and biofilm formation mediated by these pathogenic bacterial strains. Early attachment and subsequent biofilm development were retarded on surfaces with nanopores of 15-25 nm in diameter compared to surfaces with 50-100 nm pore diameter and nanosmooth surfaces. After 30 min of incubation in nutritive media, the biomass accumulation per unit surface area was 2·93 ± 1·72 µm3  µm-2 for the 15 nm, 3·49 ± 1·97 µm3  µm-2 for the 25 nm, as compared to 14·04 ± 6·39 µm3  µm-2 for the nanosmooth, 11·88 ± 9·72 µm3  µm-2 for the 50 nm and 12·09 ± 11·84 µm3  µm-2 for the 100 nm surfaces respectively. These findings suggest that anodic alumina with small size nanoscale pores could reduce the incidence of staphylococcal biofilms and infections, and shows promise as a material for a variety of medical applications and food contact surfaces. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper reports on a simple, robust and scientifically sound method to reduce attachment and biofilm formation by Staphylococcus aureus and Staphylococcus epidermidis to abiotic surfaces using a carefully designed nanoscale topography. This approach can help to reduce the incidence of staphylococcal biofilms and infections without imposing selective stresses on bacteria, thus preventing the creation of resistant strains.

The effects of combined human parathyroid hormone (1-34) and zoledronic acid treatment on fracture healing in osteoporotic rats.

UNLABELLED: Ovariectomized (OVX) rats with tibial fracture received vehicle, ZA, PTH, or ZA plus PTH treatment for 4 and 8 weeks. Bone metabolism, callus formation, and the mass of undisturbed bone tissue were evaluated by serum analysis, histology, immunohistochemistry, radiography, micro-computerized tomography, and biomechanical test. INTRODUCTION: Previous studies have demonstrated the effect of ZA or PTH on osteoporotic fracture healing. However, reports about effects of ZA plus PTH on callus formation of osteoporotic fracture were limited. This study was designed to investigate the impact of combined treatment with ZA and PTH on fracture healing in OVX rats. METHODS: Twelve weeks after bilateral ovariectomy, all rats underwent unilateral transverse osteotomy on tibiae. Animals then randomly received vehicle, ZA (1.5 µg/kg weekly), PTH (60 µg/kg, three times a week), or ZA plus PTH until death at 4 and 8 weeks. The blood and bilateral tibiae of rats were harvested for evaluation. RESULTS: All treatments increased callus formation and strength other than the control; ZA + PTH showed the strongest effects on percent bone volume (BV/TV), trabecular thickness, total fluorescence-marked callus area, and biomechanical strength. Additionally, inhibited RANKL and enhanced osteoprotegerin expression were observed in the ZA + PTH group. But no difference in bone mineral density and BV/TV of the contralateral tibiae was observed between treated groups. CONCLUSION: Findings in this study suggested an additive effect of ZA and PTH on fracture healing in OVX rats, and this additive effect was specific to callus formation, not to undisturbed bone tissue.

Involvement of microglial cells in infrasonic noise-induced stress via upregulated expression of corticotrophin releasing hormone type 1 receptor.

Infrasound is a kind of environmental noise and threatens the public health as a nonspecific biological stressor. Upregulated expression of corticotrophin releasing hormone (CRH) and its receptor CRH-R1 in the neurons of hypothalamic paraventricular nucleus (PVN) was reported to be responsible for infrasonic noise-induced stress and injuries. Recent studies revealed that CRH-R1 is expressed in activated microglial cells, lending support to the hypothesis that microglial cells may be also responsible for infrasonic noise-induced stress. In this work, we exposed Sprague-Dawley rats and in vitro cultured microglial cells to infrasound with a main frequency of 16 Hz and a sound pressure level of 130 dB for 2 h, and examined the changes in the expression of CRH-R1 at different time points after infrasound exposure by immunohistochemistry and semi-quantitative RT-PCR. We found that infrasound exposure resulted in a significant activation of microglia cells and upregulated their expression of CRH-R1 in the PVN in vivo. Upregulated expression of CRH-R1 can be blocked by antalarmin, a selective CRH-R1 antagonist. Our in vitro data further revealed that in the absence of neurons, infrasound can directly induce microglial activation and upregulate their CRH-R1 expression. These findings suggest that in addition to the PVN neurons, microglial cells are the effector cells for infrasound as well, and involve in the infrasound-induced stress through upregulated expression of CRH-R1.

Systemic treatment with strontium ranelate promotes tibial fracture healing in ovariectomized rats.

UNLABELLED: Systemic treatment with strontium ranelate (SR) was performed on ovariectomized (OVX) rats with fractured tibiae. Callus quality was assessed by radiographic, histological, micro-computerized tomography, and biomechanical examinations at 4 and 8 weeks after fracture. Results revealed that systemic applied SR promoted osteoporotic fracture healing. INTRODUCTION: Several studies have demonstrated the dual effect of SR on osteoporotic and undisturbed bone. However, reports of their effect on osteoporotic fracture healing are limited. This study was designed to investigate the effects of SR on bone regeneration in OVX rats with fractured tibiae. METHODS: Three months after being OVX, female Sprague-Dawley rats accepted bilateral osteotomy on proximal tibiae fixed with intramedullary wires and were divided into two groups: OVX and OVX + SR (625 mg/kg/day). Callus quality was evaluated at 4 and 8 weeks postfracture. RESULTS: Compared with OVX group, SR treatment significantly increased bone formation, BMD, biomechanical strength, and improved microstructural properties of the callus. The ultimate load was increased by 211.0% and 61.4% (p<0.01), and the total bone volume of callus by 74.8% and 79.3% (p<0.01) at 4 and 8 weeks postfracture, respectively. SR treatment also promoted healing progress with increased osteogenesis at 4 weeks; more mature and tightly arranged woven or lamellar bone at 8 weeks across the fracture gap in histological analysis. CONCLUSION: This study suggests that systemic treatment with strontium ranelate could promote tibial fracture healing in OVX rats.

Improved tolerance of maize plants to salt stress by arbuscular mycorrhiza is related to higher accumulation of soluble sugars in roots.

The effect of colonization with the arbuscular mycorrhizal (AM) fungus Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe on the growth and physiology of NaCl-stressed maize plants ( Zea mays L. cv. Yedan 13) was examined in the greenhouse. Maize plants were grown in sand with 0 or 100 mM NaCl and at two phosphorus (P) (0.05 and 0.1 mM) levels for 34 days, following 34 days of non-saline pre-treatment. Mycorrhizal plants maintained higher root and shoot dry weights. Concentrations of chlorophyll, P and soluble sugars were higher than in non-mycorrhizal plants under given NaCl and P levels. Sodium concentration in roots or shoots was similar in mycorrhizal and non-mycorrhizal plants. Mycorrhizal plants had higher electrolyte concentrations in roots and lower electrolyte leakage from roots than non-mycorrhizal plants under given NaCl and P levels. Although plants in the low P plus AM fungus treatment and those with high P minus AM fungus had similar P concentrations, the mycorrhizal plants still had higher dry weights, soluble sugars and electrolyte concentrations in roots. Similar relationships were observed regardless of the presence or absence of salt stress. Higher soluble sugars and electrolyte concentrations in mycorrhizal plants suggested a higher osmoregulating capacity of these plants. Alleviation of salt stress of a host plant by AM colonization appears not to be a specific effect. Furthermore, higher requirement for carbohydrates by AM fungi induces higher soluble sugar accumulation in host root tissues, which is independent of improvement in plant P status and enhances resistance to salt-induced osmotic stress in the mycorrhizal plant.

[Mechanism of apoptosis induced by squamocin in leukemia cells].

AIM: To investigate the mechanism of apoptosis of HL60 cells induced by the annonaceous acetogenin, squamocin. METHODS: Induction of apoptosis was determined through Hoechst33258 dye staining and DNA agarose gel electrophoresis. Expression of the proteins was detected using Western blot analysis. Caspase-3 activity was detected using caspase-3 kit. RESULTS: Treatment of HL-60 cells with squamocin resulted in extensive nuclear condensation, DNA fragmentation, cleavage of the death substrate poly(ADP-ribose) polymerase (PARP) and induction of caspase-3 activity. Pretreatment of HL-60 cells with caspase-3 specific inhibitor DEVD-CHO prevented squamocin-induced DNA fragmentation, PARP cleavage and cell death. Stress-activated protein kinase (SAPK/JNK) was activated after treatment with squamocin in HL-60 cells. CONCLUSION: These results suggest that apoptosis of HL-60 cells induced by squamocin require caspase-3 activation, and could be related to SAPK activation.

[Effect of AM fungi on water and nutrition status of corn plants under salt stress].

Under NaCl stress, the dry matter production of corn plants inoculated with or without arbuscular mycorrhizal (AM) fungus (Glomus mosseae) was decreased, but the decrement for non-mycorrhizal plants was 10% higher than that for mycorrhizal ones. Under salt stress condition, the dry weights of root system and aboveground part of mycorrhizal corn and its leaf water potential were higher than those of non-mycorrhizal corn, while the proline content of mycorrhizal corn was less than that of non-mycorrhizal corn. The contribution of hypha to P uptake of plants decreased from 45.3% to 42.6%, while the effect of AM fungi on plant growth increased from 30.9% to 63.5% under salt stress condition. The above-mentioned results indicated that the mechanism that AM fungi enhance the salt-resistance of corn is related with the improvements of water and P nutrition conditions. Meanwhile, it was found whether under salt stress or not, the ratio of P accumulation of root system to aboveground part of mycorrhizal corn was higher than that of non-mycorrhizal corn, indicating that the infection of AM fungi changed the P distribution pattern in plant bodies, which is beneficial to increase the salt-resistance of plants.

[Analysis of chemical constituents in the essential oil from Evodia fargesii by GC/MS].

In the essential oil from Evodia fargesii Dode, 33 constituents were identified by means of GC-MS-DS. The content of these compounds are 91.0% in the essential oil of this plant. The main compounds are eighteen terpenoids.

[Study on cultivation of Dendrobium flexicaule by habitat imitation].

Dendrobium drug, a rare medicinal herb, reproduces differcut for growth environment to lead rare resources. By studying of cultivation for many many years in Fuliu mountain area, authors had found out the cultivated method simulating the habitat of wild Dendrobium flexicaule. The results could supply a basis for its development and utilization.

Bletilla striata as a vascular embolizing agent in interventional treatment of primary hepatic carcinoma.

OBJECTIVE: To evaluate Bletilla striata as a vascular embolizing agent in interventional treatment of primary hepatic carcinoma. METHODS: We made a vascular embolizing agent from a Chinese medicinal herb, Bletilla striata (bai ji). From October 1991 to January 1995, 56 patients with primary hepatic carcinoma were treated by hepatic arterial chemoembolization with Bletilla striata angioembolus, and 50 patients with primary hepatic carcinoma who were treated by conventional gelfoam embolization served as controls. All patients were followed up for 10-48 months. RESULTS: Embolization with the Bletilla striata powder produced extensive and permanent vascular obstruction, resulting in marked shrinkage of the tumor and significant decrease of serum alpha fetoprotein (AFP) level. After embolization, less collateral circulation formed later. The treatment interval was prolonged to an average of 7 months. The 1-, 2- and 3-year survival rates were 81.9%, 44.9%, and 33.6%, respectively, with a mean survival period of 19.8 months. All the clinical parameters were better than those in the control group treated by conventional gelfoam embolization. CONCLUSION: Bletilla striata angioembolus is a good vascular embolizing agent for treating PHC.

[Embolotherapy of craniofacial tumors and vascular diseases: assessing the effect of different embolic agents].

To assess the effect of different embolic agents in curing craniofacial tumors and vascular diseases, 21 patients who were treated by intra-arterial embolization were divided into three groups: gelfoams, bletilla striatas and coils. The results showed that 8 cases of preoperative patients in gelfoams group had a good effect and bleeding were reduced during operation. One patient had vascular recanalization at 1 month after embolization with gelfoam. All patients in bletilla striatas had ideal effect. In coils group, collateral supplying arteries were found in 3 patients at once after embolization and the others had a good effect. In conclusion, craniofacial tumors and vascular diseases were fit to embolotherapy. The selection of embolic agents should be gelfoam in preoperative embolization and bletilla striata in palliative treatment There were not severe complications.

[New use of Bletilla striata as embolizing agent in the intervention treatment of hepatic carcinoma].

A Chinese herb, Bletilla striata, was used as embolizing agent in order to improve the therapeutic results of intervention treatment of liver cancer. From October 1991 to January 1995, 56 cases of hepatic carcinoma were treated with Bletilla striata by hepatic artery embolization, with conventional gelform embolization in 50 cases as control. Patients were followed-up for 10-48 months. Embolization with Bletilla striata led to extensive and permanent vascular obstruction, accompanied with marked shrinkage of tumor size and significant decrease in serum AFP levels. Collaterals were few in number and collateral circulation was established late so that the treatment intervals could be prolonged, with an average of 7 months. The 1-, 2-, and 3-year survival rate was 81.9%, 44.9% and 33.6%, respectively, with a median survival time of 19.8 months. All the clinical parameters were better than those treated with conventional gelform embolization. The results indicate that Bletilla striata is an ideal vascular embolizing agent.

[Assessment of the toxicity of TAH on the cell lines of SO-Rb50 and SO-Rb70].

PURPOSE: MTT assay was evaluated on cytoxicity for suspension growing cell lines of SO-Rb50 and SO-Rb70, and the toxicity of TAH (the total alkaloid of peqanum harmala L) on the above cell lines was assessed. METHODS: The relationships between cell number and optical density, between optical density and exposure time of MTT, and the stability of formazan crystal solution in MDSO were determined. And the toxicity of TAH on the cell lines of SO-Rb50 and SO-Rb70 in vitro with MTT assay was assessed. RESULTS: There was a direct proportional relationship between the amount of cell number and its optical density; The optical density increased gradully within 12 hours of the MTT incubation time; The stable time of the formazan crystal solved in DMSO was 11 hours. The IC50 values (micrograms/ml) of TAH on SO-Rb50 were 10.66, 4.82 respectively for 48 and 72 hours; and on SO-Rb70 were 6.38, 4.2 respectively for 48 and 72 hours. CONCLUSION: MTT assay can be used for suspension growing cell lines of SO-Rb50 and SO-Rb70: TAH has obvious toxicity to these two cell lines.

Cloning and functional characterization of a novel dopamine receptor from Drosophila melanogaster.

A cDNA clone is described that encodes a novel G-protein-coupled dopamine receptor (DopR99B) expressed in Drosophila heads. The DopR99B receptor maps to 99B3-5, close to the position of the octopamine/tyramine receptor gene at 99A10-B1, suggesting that the two may be related through a gene duplication. Agonist stimulation of DopR99B receptors expressed in Xenopus oocytes increased intracellular Ca2+ levels monitored as changes in an endogenous inward Ca2+-dependent chloride current. In addition to initiating this intracellular Ca2+ signal, stimulation of DopR99B increased cAMP levels. The rank order of potency of agonists in stimulating the chloride current is: dopamine > norepinephrine > epinephrine > tyramine. Octopamine and 5-hydroxytryptamine are not active (< 100 microM). This pharmacological profile plus the second-messenger coupling pattern suggest that the DopR99B receptor is a D1-like dopamine receptor. However, the hydrophobic core region of the DopR99B receptor shows almost equal amino acid sequence identity (40-48%) with vertebrate serotonergic, alpha 1- and beta-adrenergic, and D1-like and D2-like dopaminergic receptors. Thus, this Drosophila receptor defines a novel structural class of dopamine receptors. Because DopR99B is the second dopamine receptor cloned from Drosophila, this work establishes dopamine receptor diversity in a system amenable to genetic dissection.

Grap is a novel SH3-SH2-SH3 adaptor protein that couples tyrosine kinases to the Ras pathway.

A human cytoplasmic signaling protein has been cloned that possesses the same structural arrangement of SH3-SH2-SH3 domains as Grb2. This protein is designated Grap for Grb2-related adaptor protein. The single 2.3-kilobase (kb) grap transcript was expressed predominantly in thymus and spleen, while the ubiquitously expressed grb2 gene produced two mRNA species of 3.8 and 1.5 kb. Grap and Grb2 consist of 217 amino acids and share 59% amino acid sequence identity, with highest homology in the N-terminal SH3 domain. The GrapSH2 domain interacts with ligand-activated receptors for stem cell factor (c-kit) and erythropoietin (EpoR). Grap also forms a stable complex with the Bcr-Abl oncoprotein via its SH2 domain in K562 cells. Furthermore, Grap is associated with a Ras guanine nucleotide exchange factor mSos1, primarily through its N-terminal SH3 domain. These results show that a family of Grb2-like proteins exist and couple signals from receptor and cytoplasmic tyrosine kinases to the Ras signaling pathway.

Renal angiomyolipoma: diagnosis with B-ultrasonography, CT scanning, DSA and its interventional treatment.

From 1989, 15 cases of renal angiomyolipoma (AML) have been diagnosed by ultrasonography. CT scanning and digital subtraction angiography (DSA) at our hospital. In 8 patients with uneven hyperechoes on B-mode ultrasonography (B-US) (8/15) and 7 with low density of fat on CT scanning (7/12) accurate diagnosis was established preoperatively. DSA revealed the "berry-like" pseudoaneurysms in the arterial phase (14 cases), the defined lucent area in the nephrogram phase (10 cases) and the "onion-peel appearances" during venous phases (8 cases), correct diagnosis was achieved in all patients. 8 cases were surgically treated and 7 treated by subselective embolization of renal artery. Effects in all cases were good. The diagnostic value of B-US, CT scanning, DSA and interventional treatment of AML was discussed. It was believed that the diagnosis with DSA was a technique with high specificity, and embolization therapy was simple and effective for AML.

[Comparative study on the long-term effect of permanent embolization of hepatic artery with Bletilla striata in patients with primary liver cancer]. / Permanente Chemoembolisation des Leberkarzinoms mit dem chinesischen Heilmittel Bletilla striata: Langzeitergebnisse im Vergleich mit Lipiodol-MMC-Gelfoam.

From May, 1990 to Sept., 1993 106 cases of primary liver cancer were treated by temporary or permanent hepatic arterial embolization with Bletilla striata powders (permanent, 56 cases) or gelfoam powders (temporary, 50 cases) under controlled technical conditions, in term of degree of tumor necrosis, revascularization of tumor, and long-term effect, the results of Bletilla striata group were better as compared with those of gelfoam group. The 1, 2 and 3 year survival rates were 44.9%, 33.6% and 33.6% in Beltilla striata group while the rates were 48.9%, 31.1% and 16.0% in gelfoam group, suggesting that Bletilla striata is superior to gelfoam as an embolizing agent for hepatic carcinoma.