Studies on the Proton Pumping Activity of H^+-ATPase in Tonoplast Vesicles of Populus euphratica

Tonoplast-enriched vesicles were prepared from suspension-cultured Populus euphratica Oliv. cells by differential centrifugation and discontinuous sucrose density gradient centrifugation. The properties of the proton pumping activity of H+-ATPases in tonoplast vesicles were studied by acridine orange fluorescent quenching measured at 22 degreesC. The proton pumping activity of ATPase was ATP-dependent with apparent Michaelis-Menten Constant (K-m) for ATP about 0.65 mmol/L. The optimal pH for H+-ATPases activity was 7.5. The proton pumping activity of H+-ATPase could be initiated by some divalent cations, Mg2+ being highly efficient, much more than Fe2+; and Ca2+, Cu2+ and Zn2+ were inefficient under the experimental condition. The proton translocation could be stimulated by halide anions, with potencies decreasing in the order Cl- > Br- > I- > F-. The proton pumping activity was greatly inhibited by N-ethylmaleimide (NEM), N, N'-dicyclohexylcarbodiimide (DCCD), NO3- and Bafilomycin A(1), but not by orthovanadate and azide. These results demonstrated that the H+-ATPase in the tonoplast of Populus euphratica belonged to vacuolar type ATPase. This work was the first time that tonoplast-enriched vesicles were isolated from Populus euphratica cells.

The Relationship Between Senescence and Ca^(2+)-ATPase Activity of Microsomal Membrane and Lipid Peroxidation in Harvested Peach Fruit

Peach fruit easily soften and have a short storage time at normal temperature. In this study, peach fruit （Prunus persica sieb et Zucc cv. Yingqing） were picked and stored at 25 and 4℃ to investigate the senescence in correlation with Ca^2＋- ATPase activity of microsomal membrane and lipid peroxidation during ripening and senescence. In comparison with that stored at 25~C, the fruit stored at 4℃ exhibited a higher flesh firmness, lower respiration rate, and generated the late bigger peak value of Ca^2＋-ATPase activity as well as maintained the higher activity of the enzyme. Meanwhile, the lower levels of super oxygen radical （O2^-.） production and content of malondialdehyde （MDA）, a product of membrane lipid peroxidation were observed. Sodium orthovanadate （SO） and erythrosin B （EB）, as Ca^2＋-ATPase inhibitors, could stimulate the respiration rate. The results suggested that the slower senescence rate of peach fruit was closely related to the higher peak value and longer duration of Ca^2＋-ATPase activity in microsomal membrane, with the slighter membrane lipid peroxidation and lower O2^-. production rate.

The investigation for the relationship among serum leptin, erythrocyte membrane Ca^(2+)-ATPase activity and hypertensive disorder complicating pregnancy

Objective： To study the significance of Leptin and the activity of erythrocyte membrane Ca^2＋-ATPase（EMCA） in the development of hypertensive disorder complicating pregnancy. Methods： Radioimmunoassay was used to test the level of serum Leptin, and the activity of EMCA was determined chemically in 38 pregnant women with hypertensive disorder complicating pregnancy and 36 normotensive pregnant women. Results： The level of serum Leptin in hypertensive disorder complicating pregnancy（gestational hypertension： 13.76 ± 3.46 ng/ml; preeclampsia： 15.76 ± 5.47 ng/ml; eclampsia： 18.32 ± 6.38 ng/ml）was significantly higher than that in normotensive pregnant women （11.33 ± 2.93 ng/ml） ,respectively. The average EMCA activity of patients with hypertensive disorder complicating pregnancy （gestational hypertension： 1.65 ± 0.24 μmol·pi/mg.h ; preeclampsia： 1.37 ± 0.19 μ mol·pi/mg·h;eclampsia：1.12 ± 0.14 μmol·pi/mg·h） was significantly lower than that of normotensive pregnant women（1.83 ± 0.38 μ mol·pi/mg·h）,respectively. There was a negative correlation between the level of serum Leptin and the activity of RMCA in hypertensive disorder complicating pregnancy （r = -0.63）. Conclusion： Inhibition of EMCA activity of erythrocyte in hypertensive disorder complicating pregnancy may increase cytoplasmic free calcium, which contributes to the development of hypertensive disorder complicating pregnancy. The negative correlation between the level of serum Leptin and the activity of EMCA, also suggested that serum Leptin and the activity of EMCA may play a role in the development of hypertensive disorder complicating pregnancy.

Effects of Salinity Fluctuation Frequency on the Osmolarity,Na^+-K^+-ATPase Activity and HSP70 Expression in Juvenile Chinese Shrimp,Fenneropenaeus chinensis

Experiments were conducted to examine the effects of salinity fluctuation frequency on the osmolarity, Na^＋-K^＋-ATPase activity and HSP70 of Chinese shrimp Fenneropenaeus chinensis ruth initial wet body weight of 1.460g±0.091 g. The salinity in the control group （DO） was 28 throughout the experiment, whereas treatments D2, D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8d, respectively. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8d, respectively, decreased abruptly to salinity 24, lasted for another 2 d, and then was raised to its initial value 28. This was a complete salinity fluctuation cycle that afterwards repeated itself. After 32 days, the osmolarity in treatments D2, D4, D6 and D8 was significantly lower than that in treatment DO （P〈0.05）. There were significant differences in both muscle and eyestalks HSP70 expression among groups. The HSP70 expressions in muscle and eyestalks in group D4 were 61.4% and 57.0% higher, respectively, than that in the control group DO （P〈0.05）. There were, however, no significant differences in gill or hepatopancreas Na^＋-K^＋-ATPase activity between the treatments and the control.

Effects of salinity fluctuation amplitudes on growth,osmolarity,Na^+-K^+-ATPase activity and Hsp70 of juvenile Chinese shrimp Fenneropenaeus chinensis Osbeck

The effects of various salinity fluctuation amplitudes （2, 4, 6 and 8） on the growth, osmolarity, Na＋-K＋-ATPase activity and Hsp70 of juvenile Fenneropenaeus chinensis cultured in seawater with a salinity of 20 were studied. The results show that weight gain in the salinity fluctuation treatments was better than that in control; in particular, the weight gain of treatments S4 and SO, at 231.8% and 196.3%, respectively, was significantly different （P〈0.05）. The hemolymph osmolarity of treatments SO, S2, S4, S6 and S8 was 635.4, 630.8, 623.6, 614.4 and 600.3 mOsm/kg, respectively, and decreased with increasing salinity fluctuation amplitude. The level of Na＋-K＋-ATPase activity in gills ofE chinensis was higher than that in hepatopancreas, but there were no significant differences among all treatments, either in gills or hepatopancreas （P〉0.05）. The relative level of Hsp70 in treatment $4 was 48.4% and 40.4% higher than control in muscle and eyestalks, respectively, with the highest values being recorded under a salinity fluctuation amplitude of 4.

EFFECTS OF CALDESMON, CALPONIN, AND TROPOMYOSIN ON THE MG^(2+)-ATPase ACTIVITIES OF SMOOTH MUSCLE MYOSIN

Objective To test whether in the absence of actin, actin-binding proteins such as caldesmon, calponin, and tropomyosin interact with the myosin of unphosphorylation, Ca 2+ -dependent phosphorylation (CDP), and Ca 2+ -independent phosphorylati-on (CIP) and stimulate myosin Mg 2+ -ATPase activities. Methods Mg 2+ -ATPase activities were measured to evaluate the effects of caldesmon, calponin, and tropomyosin on the myosin in unphosphorylation, CDP by myosin light chain kinase (MLCK), and CIP by MLCK. Results (1) At different incubation-time, i.e., 5, 10, 20, 40, and 60 minutes, the highest Mg 2+ -ATPase activity was ob-served when myosin was in the state of CDP, the medium was CIP of myosin, and the lowest was the unphosphorylated myosin. (2) In the absence of caldesmon, calponin, and tropomyosin, the Mg 2+ -ATPase activities from high to low were in the following order: CDP, CIP, and unphosphorylated myosin. However, in the presence of caldesmon, calponin, and tropo-myosin, the order of relative value of Mg 2+ -ATPase activities from high to low was unphosphorylated, CIP, and CDP of myosin respectively compared to the corresponding controls. Conclusions The results propose that caldesmon, calponin, and tropomyosin are capable of stimulating Mg 2+ -ATPase activity of smooth muscle myosin in Ca 2+ -independent manner, since Ca 2+ is not obligating for the stimulating effects of the three proteins. The common characteristic of the three proteins is that when myosin activities are low, their activations are relatively strong and this property might be involved in smooth muscle tension keeping.

Ethanolic extract of Abrus precatorius leaves protected against mercury chloride-induced hepatocellular damage and cerebral Na+/K+-ATPase dysfunction in wistar rats

Plants are sources of medicinal compounds,and they play a crucial role in human health maintenance.Abrus precatorius is one of the important medicinal plants that have been alleged for their medicinal properties.This research unraveled the pharmacological effect of ethanolic extract of Abrus precatorius on lipid peroxidation,liver parameters,and Na^(+)/K^(+)-ATPase activity in HgCl_(2) treated wistar rats.Twenty-four(24)albino wistar rats weighing between 150-200 g were distributed into four groups of 6 animals each.Group A(control)received normal saline(0.9% NaCl),group B received 400 mg/kg of the extract only,group C received 4 mg/kg HgCl_(2) only,and group D received 400 mg/kg of extract+4 mg/kg of HgCl_(2).The treatment lasted for two weeks,and the animals were sacrificed on the 15th day.The blood,brains and livers were collected and used for assay of lipid peroxidation,liver function,and sodium pump activity.The results of liver function test revealed an elevated(P<0.05)level of serum aspartate transaminase,alanine transaminase,alkaline phosphatase,and total bilirubin in the group that received HgCl_(2) only(group C)when compared with the normal control(group A)that received normal saline only.However,the administration of extract in group D led to a marked(P<0.05)reduction in the activities of these enzymes and the level of total bilirubin when compared to the negative control(group C).On the contrary,HgCl_(2) caused a significant(P<0.05)reduction in serum total protein and albumin levels,but the extract reversed the effect of HgCl_(2) by elevating their concentrations.Nonetheless,the effect elicited by this extract is comparable to group A which received normal saline.Moreso,the result of lipid peroxidation revealed that HgCl_(2) treatment caused a marked(P<0.05)increase in the formation of lipid peroxidation adducts in both liver and brain homogenates in group C.On the contrary,administration of Abrus precatorius extract profoundly(P<0.05)inhibited HgCl_(2)-induced lipid peroxidation in group D.In addition,HgCl_(2) inhibited the activity of cerebral Na^(+)/K^(+)-ATPase,but the extract restored normalcy by increasing the activity of the enzyme in group D.Consequently,the results obtained justify the traditional use of Abrus precatorius and suggest that Abrus precatorius leaves may be used for management of liver diseases,oxidative stress-linked diseases and some neurodegenerative ailments.

盐胁迫下盐地碱蓬叶片液泡膜H^+-ATPase H亚基的克隆与表达分析

利用EST随机挑取克隆测序的方法从盐地碱蓬叶片cDNA文库中分离得到了盐地碱蓬V-H+-ATPase H亚基cDNA序列,并进行了H、c亚基基因表达及V-H+-ATPase活性分析.结果表明,H亚基基因全长1 969 bp,包括100 bp的5′-非编码区和471 bp的3′-非编码区.开放阅读框为1 398 bp,编码465个氨基酸残基,分子量约52.8 kD.N orthern杂交分析表明盐胁迫明显诱导了H亚基表达,而且盐胁迫下H、c亚基及V-H+-ATPase活性存在协同作用.这些结果表明盐胁迫下H和c亚基基因上调及V-H+-ATPase活性的增加为N a+区隔化到液泡中提供了质子驱动力.

A novel phenol-bound pectic polysaccharide from Decalepis hamiltonii with multi-step ulcer preventive activity

AIM: To investigate H+, K+-ATPase inhibition, anti-H pylori , antioxidant, and the in vivo antiulcer potential of a pectic polysaccharide from Swallow root (Decalepis hamiltonii; SRPP). METHODS: SRPP, with known sugar composition [rhamnose: arabinose: xylose: galactose in the ratio of 16:50:2:32 (w/w), with 141 mg/g of uronic acid] was examined for anti-ulcer potency in vivo against swim/ ethanol stress-induction in animal models. Ulcer index, antioxidant/antioxidant enzymes, H+, K+-ATPase and gastric mucin levels were determined to assess the anti- ulcer potency. Anti-H pylori activity was also determined by viable colony count and electron microscopic studies. RESULTS: SRPP, containing phenolics at 0.12 g GAE/g, prevented stress-induced gastric ulcers in animal models by 80%-85%. Down regulation of gastric mucin 2-3 fold, antioxidant/antioxidant enzymes and upregulation of 3 fold of H+, K+-ATPase in ulcerous animals were normalized upon treatment with SRPP. Histopathological analysis revealed protection to the disrupted gastric mucosal layer and epithelial glands. SRPP also inhibited H+, K+-ATPase in vitro, at an IC50 of 77 μg/mL as opposed to that of 19.3 μg/mL of Lansoprazole and H pylori growth at Minimum Inhibitory Concentration (MIC) of 150 μg/mL. In addition, free radical scavenging (IC50-40 μg/mL) and reducing power (3200 U/g) activities were also observed. CONCLUSION: SRPP, with defined sugar composition and phenolics, exhibited multi-potent free radical scavenging, antioxidant, anti-H pylori, inhibition of H+, K+-ATPase and gastric mucosal protective activities. In addition, SRPP is non-toxic as opposed to other known anti-ulcer drugs, and therefore may be employed as a potential alternative for ulcer management.

Effects of Aluminum on ATPase Activity and Lipid Composition of Plasma Membranes from Wheat Roots

The effects of aluminum on ATPase activity and lipid composition of the plasma membranes isolated from root tips of Al-tolerant (Altas 66) or Al-sensitive (Scout 66) cultivar of Triticum aestivum L. was assayed. The results showed that both cultivars had similar changes in H+-ATPase and Ca2+-ATPase activities after aluminum treatment. Exposure of both cultivars to 20 and 100 (mol/L aluminum for 5 d significantly decreased the activities of Ca2 + -ATPase of plasma membranes. The activities of H+-ATPase in plasma membrane increased under 20 μmol/L aluminum and decreased at 100 u.mol/L aluminum. With aluminum treatment, the PL content of plasma membrane decreased, but GL content increased. The ratio of PL to GL decreased more distinctly in Scout 66 than that in Altas 66. Treated with 20 and 100 μmol/L aluminum, linolenic acid content and the index of unsaturated fatty acids decreaced greatly in Scout 66, but the index of unsaturated fatty acids in Altas 66 increased slightly.

THE CHARACTERIZATION OF MYOSIN LIGHT CHAIN PHOSPHORYLATION BY THE CONSTITUTIVELY ACTIVE FRAGMENT OF MLCK

Objective.To study the exact effects and characteristics of a constitutively active myosin light chain kinase fragment (MLCKF)on Ca 2+ -CaM dependent phosphorylation of myosin light chains(CDPM)of smooth muscle.Methods.Proteolysis of myosin light chain kinase(MLCK)by trypsin was used to obtain the con-stitutively active fragment of MLCK;Western blot was applied to demonstrate the homogeneity of typsin-digested MLCK fragments and intact MLCK.The phosphorylation of myosin light chains was detected us-ing SDS-PAGE and Scoin Image Software,and myosin Mg 2+ -ATPase activities were measured using spec-trophotometry to observe the characteristics of MLCKF in phosphorylating myosin light chains and stimu-lating myosin Mg 2+ -ATPase activities compared with intact MLCK.Results.Our assay demonstrated that MLCKF had a specific activity on CDPM.We also found that the extent of CDPM by MLCKF was greater than that by intact MLCK at different concentrations,differ-ent incubation time and different incubation temperatures.CDPM by MLCKF was also less influenced by the change of ionic strength of KCl than CDPM by intact MLCK.Mg 2+ -ATPase activities of phosphorylat-ed myosin light chains by MLCKF were higher than those by MLCK at different concentrations and dif-ferent reaction time.These differences were statistically significant ( P<0.01, P<0.05).Conclusion.MLCKF not only possessed a specific activity on CDPM but also was more efficient than MLCK in phosphorylating myosin light chains and stimulating myosin Mg 2+ -ATPase activities.Our results suggested that MLCKF possibly plays a certain role in smooth muscle contraction-relaxation cycle.

Dynamic functional and ultrastructural changes of gastric parietal cells induced by water immersion-restraint stress in rats

AIM： To investigate the dynamic functional and ultrastructural changes of gastric parietal cells induced by water immersion-restraint stress （WRS） in rats.METHODS： WRS model of Sprague-Dawley （SD） rats was established. Fifty-six male SD rats were randomly divided into control group, stress group and post-stress group. The stress group was divided into 1, 2 and 4 h stress subgroups. The post-stress group was divided into 24, 48 and 72 h subgroups. The pH value of gastric juice, ulcer index （UI） of gastric mucosa and H^＋, K^＋- ATPase activity of gastric parietal cells were measured. Ultrastructural change of parietal cells was observed under transmission electron microscope （TEM）.RESULTS： The pH value of gastric juice decreased time-dependently in stress group and increased in post-stress group. The H^＋, K^＋-ATPase activity of gastric parietal cells and the UI of gastric mucosa increased time-dependently in stress group and decreased in poststress group. Compared to control group, the pH value decreased remarkably （P = 0.0001）, the UI and H^＋, K^＋- ATPase activity increased significantly （P = 0.0001, P = 0.0174） in 4 h stress subgroup. UI was positively related with stress time （r = 0.9876, P 〈 0.01） but negatively with pH value （r = -0.8724, P 〈 0.05）. The parietal cells became active in stress group, especially in 4 h stress subgroup, in which plenty of intracellular canalicular and mitochondria were observed under TEM. In post-stress group, the parietal cells recovered to resting state.CONCOUSION： The acid secretion of parietal cells is consistent with their ultrastructural changes during the development and healing of stress ulcer induced by WRS and the degree of gastric mucosal lesions, suggesting gastric acid play an important role in the development of stress ulcer and is closely related with the recovery of gastric mucosal lesions induced by WRS.

Kinetics of the H＋-ATPase from Dry and 5-Hours- Imbibed Maize Embryos in Its Native, Solubilized, and Reconstituted Forms

Membranes undergo recovery upon rehydration in seed germination. Previous work has described that the plasma membrane H＋-ATPase from maize embryos adopts two different forms at 0 and 5 h of imbibition. We investigated how the kinetics of these two forms could be affected by alterations in the plasma membrane （PM）. In comparison to the O-h, PMs from the 5-h imbibed embryos showed changes in glycerophospholipid composition, decrease in leakage, and increase in fluidity. Kinetics of the PM H^-ATPase from 0 and 5-h imbibed embryos showed negative cooperativity, With the removal of the membrane environment, the activity of the enzymes shifted to a more complex kinetics, displaying two enzyme components. Lipid reconstitution produced one component with positive cooperativity. In all cases, enzymes from 0 and 5-h imbibed embryos presented similar kinetics with some quantitative differences. These results indicate that the two enzyme forms have the potential ability to respond to changes in the membrane enyiror^rpent, but the fact that they do not show differences in the native membranes at 0 or 5 h implies that modifications in the membrane are not drastic enough to alter their kinetics, or that they are able to preserve their boundary lipids or associated proteins and thus retain the same kinetic behavior.

Synthesis and Crystal Structure of (E)-2-phenyl-6,7dihydrobenzofuran-4(5H)-one O-cyanomethyl Oxime

A pair of E/Z-isomers of 2-phenyl-6,7-dihydrobenzofuran-4（5H）-one O-cyanomethyl oxime,C16H14N2O2,as potential drugs for treating peptic ulcer and other acid-related diseases have been synthesized and characterized by IR,MS and NMR spectra.Meanwhile,the crystal of IIIa was obtained and determined by X-ray single-crystal diffraction.Crystal data： monoclinic system,space group P21 /c,a = 8.423（8）,b = 19.596（16）,c = 8.770（8）,β = 107.750（12）°,V = 1379（2）3,Z = 4,F（000） = 560,Dc = 1.283 g/cm3,μ = 0.086 mm 1,R = 0.0681 and wR = 0.2029 for 14472 independent reflections（Rint = 0.0782） and 2428 observed ones（I 2σ（I））.

Changes in growth and osmoregulation during acclimation to saltwater in juvenile Amur sturgeon Acipenser schrenckii

We evaluated the ability of juvenile Amur sturgeon (Acipenser schrenckii) to osmoregulate and grow in saltwater. Hatchery-reared juveniles (mean weight 106.8 g, 5-month old) were transferred from freshwater to 10, 20, and 25 salinity saltwater over a period of 20 d. We measured the growth, serum osmolality, ion concentrations, and Na+/K+-ATPase activity. In addition, we prepared samples of gill tissue to quantify morphological changes in gill ultrastructure. Rearing in up to 25 saltwater for 30 d had no significant effect on growth. Similarly, serum osmolality and ion concentrations were similar to levels reported in other teleosts following acclimation to saltwater. Serum osmolality and Na+, Cl- concentrations increased significantly with the initial increase in salinity. Afterwards, levels tended to stabilize and then decrease. Serum K+ levels did not change during acclimation to saltwater. Gill Na+/K+-ATPase activity increased initially as salinity was increased. However, the activity later decreased and, finally stabilized at 3.7±0.1 μmol Pi/mg·prot·h in 25 saltwater (1.6 times higher than the level in those in freshwater). In fish that were held only in freshwater, the chloride cells were located in the interlamellar regions of the filament and at the base of the lamella. Following acclimation to 25 saltwater for 30 d, the number and size of chloride cells increased significantly. Our results suggest that juvenile Amur sturgeon is able to tolerate, and grow in, relatively high concentrations of saltwater.

The Special Feature of Calponin on Myosins Phosphorylated by MLCK and PKA Respectively

Objective: To reveal the special feature of calponin (CaP) on myosins of different states. Methods: Myosin phosphorylation determination, myosin Mg^(2+)-ATPase measurement and protein binding assay were used in this study. The lowest CaP/myosin ratio used in the assay was 1/10000(mol/mol), which was 10 thousands-fold lower than the CaP/myosin ratio used in previous studies. Results: In the absence of actin, micro-amount of calponin (MAC) stimulated the Mg^(2+)-ATPase activities of myosin in different states slightly but significantly; and more importantly, MAC significantly increased the precipitations of unphosphorylated myosin, Ca^(2+)-dependently and independently phosphorylated myosins by MLCK but not the myosin phosphorylated by PKA. Conclusion: MAC has a high efficient and selective effect on myosin in the absence of actin.

Toxicity Effects of 1,4-Dinitrobenzene on Oryzias Latipes

Dinitrobenzene(1,4-DNB) is one of the preferential toxicants stipulated by OECD and EPA of USA. With Oryzias latipes, which were chosen as the acceptors according to the international standard, the acute, subacute and accumulative toxicity experiments were conducted. The threshold mass concentration of O.latipes was 1.360 mg/L. The subacute experiment showed that O.latipes were hardly affected when the mass concentration of 1,4-DNB was 0.0292 mg/L, but were significantly affected when the mass concentration of 1,4-DNB was 0.0420, 0.0608 or 0.0875 mg/L in contrast to the control sample(P<0.05). The accumulative rate(K) was 1.21. This shows that there are significant accumulative actions of 1,4-DNB in fish. When the mass concentration of 1,4-DNB is in the range of 0.032_0.320 mg/L, the higher the 1,4-DNB concentration, the more strongly the Na +/K +-ATPase is inhibited. The calculated IC_ 50 values in gill, kidney and liver of O. latipes are 0.128, 0.132 and 0.428 mg/L, respectively.

Effect of Dietary Nitrate on Force Production and Sarcoplasmic Reticulum Ca²⁺Handling in Rat Fast-Twitch Muscles Following Eccentric Contraction

Impaired excitation-contraction coupling occurs in eccentric contraction (ECC)-induced damaged muscles. It has been suggested that sarcoplasmic reticulum (SR) is susceptible to damage in the overstretched regions possibly marking the basis of excitation-contraction coupling damage. Recent studies have shown that dietary nitrate supplementation enhances SR function in fast-twitch muscles. In this study, we aimed to investigate whether dietary nitrate supplementation can alleviate a decline in muscle contractile properties and SR function following ECC. To this end, force production, Ca2+ uptake, Ca2+ release, and Ca2+-ATPase activity of the SR were examined in rat fast-twitch muscles immediately following ECC for 200 repetitions. In comparison with contralateral resting muscles, nitrate supplementation for up to 3 days resulted in an obvious decline in force production. However, there were no differences in terms of force production between 6-day nitrate-treated and contralateral muscles. Similar to the observations regarding force production, the SR Ca2+ release rate changed from an obvious decrease following the 0- and 3-day dietary nitrate supplementation to no difference following the 6-day nitrate supplementation. In contrast, ECC decreased the Ca2+-ATPase activity and Ca2+ uptake rate, irrespective of the period of dietary nitrate supplementation. Overall, these results indicate that dietary nitrate supplementation can alleviate ECC-related decreases in force production mediated through inhibited reductions in the SR Ca2+ release function.

Experimental Study on Mechanical Vibration Massage for Treatment of Brachial Plexus Injury in Rats

Objective: To investigate the curative effect of the self-made mechanical vibration massage instrument for treatment of brachial plexus injury in rats and to explore its mechanism. Methods: Brachial plexus injury models were made in 144 Wistar rats and one week after natural healing of the wound, they were randomly divided into 3 groups, mechanical vibration treatment group (MV group), nerve growth factor treatment group (NGF group) and model group, 48 rats in each group. Then again, the each group was randomly divided into 4 subgroups, 7-day group, 14-day group, 21-day group and 28-day group, 12 rats in each subgroup. The MV group were treated by mechanical vibration at acupoints on three-yang and three-yin channels of the hand with the mechanical vibration massage instrument; The NGF group were treated with injection of NGF into musculus pectoralis major on the affected side; And the model group were normally fed with no treatment. After treatment for 7, 14, 21 and 28 days, the diameter of both forelimbs were measured, the electrophysiological examination on the brachial plexus in vitro and the ultrastructure observation with electron microscope on the affected side were carried out, the motor nerve conduction velocity (MNCV) and motor nerve action potential (MNAP) of the brachial plexus on the affected side, NGF content of submaxillary gland as well as muscular Na+, K+-ATPase activity were determined respectively. Results: The different rates of the forelimb diameter in the MV group and the NGV group on the 14th d, 21st d and 28th d were better than those in the model group (P<0.05 or P<0.001), and in the MV group were better than those in the NGF group on the 21st d and the 28th d (P<0.05). MNCV in the MV group and the NGV group on the 21st d and 28th d was better than that in the model group (P<0.05 or P<0.001), and in the MV group was better than that in the NGF group on the 28th d (P<0.05). MNAP in the MV group and the NGV group on the 14th d, 21st d and 28th d was better than that in the model group (P<0.05 or P<0.001), and in the MV group was better than that in the NGF group on the 21st d and 28th d (P<0.05). The NGF mean gray index of submaxillary gland in the model group was higher than that in the MV group and the NGF group on the 7th d (P<0.05); in the NGF group and the model group was higher than that in the MV group on the 14th d (P<0.05); and in the NGF group and the MV group was higher than that in the model group on the 21st d and 28th d (P<0.05). Na+, K+-ATPase activity in the model group and the MV group was higher than that in the NGF group (P<0.05) on the 14th d, and in the MV group was higher than that in the model group on the 28th d (P<0.05). Conclusion: As compared with the NGF group and the model group, mechanical vibration treatment can effectively accelerate repair of injured brachial plexus, slow down atrophy of skeletal muscle, and promote secretion of NGF in submaxillary gland.

Highly sensitive detection of NT-proBNP by molecular motor

FoF1-ATPase is an active rotary motor,and generates three-ATP for each rotation.At saturated substrate concentration,the motor can achieve about 103 r.p.m,which means one motor can generate about 105 ATP molecules during 30 min.Here,we constituted a novel nanodevice with a molecular rotary motor and a“battery”,FoF1-ATPase and chromatophore,and presented a novel method of sandwich type rotary biosensor based on εsubunit with one target-to-one motor,in which one target corresponds 105 ATP molecules as detection signals during 30 min.The target such as NT-proBNP detection demonstrated that this novel nanodevice has potential to be developed into an ultrasensitive biosensor to detect low expressed targets.