Apple rootstocks with different phosphorus efficiency exhibit alterations in rhizosphere bacterial structure.

AIMS: The purpose of this study was to select phosphorus-efficient apple rootstocks under phosphorus deficiency and to reveal the effects of different apple rootstocks on the rhizosphere bacterial community. METHODS AND RESULTS: We used 83 hybrid lines of Malus robusta Rehd. × Malling 9 (M.9) to investigate their physiological traits and the phosphorus deficiency phenotypes of leaves in response to phosphorus deficiency (0·1 mmol l-1 PO4 3- ). All the plants were cultivated in pots in the greenhouse and watered using drip irrigation. In accordance with the results of investigation, we selected the phosphorus-efficient hybrid lines (PE) and the phosphorus-inefficient hybrid lines (PI) to research their root morphology and root hairs (RH). In addition, we used Illumina MiSeq sequencing to determine the bacterial community of the rhizosphere from different rootstocks. The results showed that the PE plants had better growth characteristics and stronger root plasticity than that of the PI plants, and phosphorus deficiency can stimulate the RH growth of PE plants. There was no significant difference in the rhizosphere bacterial diversity, but we found that the bacterial community structure was significantly different at the genus levels; in addition, 89 genera were found to have significant differences between PE and PI plants, especially Bacillus. The PE rhizosphere had more abundant Bacillus compared to the PI. High positive Pearson correlations with the phosphorus concentration in the plantlets of apple rootstocks were detected for the bacterial genera Bacillus (r: 0·776). CONCLUSIONS: The phosphorus-efficient apple rootstocks adapted to phosphorus deficiency by shaping the root morphology. Notably, different apple rootstocks showed alteration of the microbes in rhizosphere. SIGNIFICANCE AND IMPACT OF THE STUDY: This study can provide the materials for exploring the mechanism of apple rootstock phosphorus absorption. In accordance with the different bacterial community compositions, we can develop the inoculants to promote nutrient uptake.

[Effect of yiqi jianpi drugs on apoptosis and relevant gene expression in cultured vascular smooth muscle cell].

OBJECTIVE: To explore the effect of Yiqi Jianpi (YQJP, supplementing Qi and invigorating Spleen) recipe on vascular smooth muscle cells (VSMC) apoptosis and it's mechanisms. METHODS: VSMC apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nicked labeling (TUNEL) analysis, transmission electron microscope (TEM), flow cytometry and DNA agarose electrophoresis. The expression activity of bcl-2, c-myc and p53 genes were determined using Northern blotting. RESULTS: The typical characteristics of apoptotic VSMC was observed following treatment by the YQJP drug serum. The DNA extracted from VSMC revealed a ladder pattern in electrophoresis. Flow cytometric analysis revealed G0/G1 stage cell blocking phenomena, the hypodiploid apoptotic cell increased, displayed typical peak of apoptosis, the rate of cell apoptosis and YQJP serum concentration were in a dose-dependent manner. The result of hybridization showed that 30% of YQJP serum could inhibit apoptotic gene bcl-2 expression, upregulated the level of apoptogenous gene c-myc and p53 mRNA. CONCLUSION: VSMC apoptosis could be induced by YQJP recipe and its mechanism might be through affecting the apoptosis related gene expression activity.

[Wavelet application to reduction of stimulus artifact in transient evoked otoacoustic emissions testing].

Otoacoustic emissions(OAEs) are presently considered as an objective clinical test for assessing the integraity of peripheral hearing. Owing to the adding of the reflex waves of the stimuli, recordings of emissions evoked in response to transient stimuli are contaminated by an initial artifact, which inhibits the examination of high frequency cochlear responses that have short latencies. So OAEs testing is of serious limitation in most clinical environments. We propose an artifact reduction technique based on discrete wavelet transform pre-processing method and demonstrate empirically that the method not only improves artifact reduction but also enhances signal-to-noise ratio in the response region.

cDNA cloning, tissue distribution, and substrate characteristics of a cis-Retinol/3alpha-hydroxysterol short-chain dehydrogenase isozyme.

We report here a mouse cDNA that encodes a 316-amino acid short-chain dehydrogenase that prefers NAD+ as its cofactor and recognizes as substrates androgens and retinols, i.e. has steroid 3alpha- and 17beta-dehydrogenase and cis/trans-retinol catalytic activities. This cis-retinol/androgen dehydrogenase type 2 (CRAD2) shares close amino acid similarity with mouse retinol dehydrogenase isozyme types 1 and 2 and CRAD1 (86, 84, and 87%, respectively). CRAD2 exhibits cooperative kinetics with 3alpha-adiol (3alpha-hydroxysteroid dehydrogenase activity) and testosterone (17beta-hydroxysteroid dehydrogenase activity), but Michaelis-Menten kinetics with androsterone (3alpha-hydroxysteroid dehydrogenase activity), 11-cis-retinol, all-trans-retinol, and 9-cis-retinol, with V/K0.5 values of 1.6, 0.2, 0.1, 0.04, 0.005, and not saturated, respectively. Carbenoxolone (IC50 = 2 microM) and 4-methylpyrazole (IC50 = 5 mM) inhibited CRAD2, but neither ethanol nor phosphatidylcholine had marked effects on its activity. Liver expressed CRAD2 mRNA intensely, with expression in lung, eye, kidney, and brain (2.9, 2, 1.6, and 0.6% of liver mRNA, respectively). CRAD2 represents the fifth isozyme in a group of short-chain dehydrogenase/reductase isozymes (retinol dehydrogenases 1-3 and CRAD1), closely related in primary amino acid sequence (approximately 85%), that are expressed in different quantities in various tissues, have different substrate specificities, and may serve different physiological functions. CRAD2 may alter the amounts of active and inactive androgens and/or convert retinols into retinals. These data expand insight into the multifunctional nature of short-chain dehydrogenases/reductases and into the enzymology of steroid and retinoid metabolism.

cDNA cloning and characterization of a cis-retinol/3alpha-hydroxysterol short-chain dehydrogenase.

We report a mouse cDNA that encodes a 317-amino acid short-chain dehydrogenase which recognizes as substrates 9-cis-retinol, 11-cis-retinol, 5alpha-androstan-3alpha,17beta-diol, and 5alpha-androstan-3alpha-ol-17-one. This cis-retinol/androgen dehydrogenase (CRAD) shares closest amino acid similarity with mouse retinol dehydrogenase isozymes types 1 and 2 (86 and 91%, respectively). Recombinant CRAD uses NAD+ as its preferred cofactor and exhibits cooperative kinetics for cis-retinoids, but Michaelis-Menten kinetics for 3alpha-hydroxysterols. Unlike recombinant retinol dehydrogenase isozymes, recombinant CRAD was inhibited by 4-methylpyrazole, was not stimulated by ethanol, and did not require phosphatidylcholine for optimal activity. CRAD mRNA was expressed intensely in kidney and liver, in contrast to retinol dehydrogenase isozymes, which show strong mRNA expression only in liver. CRAD mRNA expression was widespread (relative abundance): kidney (100) > liver (92) > small intestine (9) = heart (9) > retinal pigment epithelium and sclera (4.5) > brain (2) > retina and vitreous (1.6) > spleen (0.7) > testis (0.6) > lung (0.4). CRAD may catalyze the first step in an enzymatic pathway from 9-cis-retinol to generate the retinoid X receptor ligand 9-cis-retinoic acid and/or may regenerate dihydrotestosterone from its catabolite 5alpha-androstan-3alpha,17beta-diol. These data also illustrate the multifunctional nature of short-chain dehydrogenases and provide a potential mechanism for androgen-retinoid interactions.

[Protective effect of puerarin on acute cerebral ischemia in rats].

Acute incomplete brain ischemia in rats was induced by bilateral carotid artery occlusion(BCAO). BCAO results in severe suppression of EEG and increase of brain water content. Left middle cerebral artery occlusion(MCAO) in rats by electrical coagulation results in increase of brain water content of ipsilateral hemisphere and contralateral hemiparesis. The typical ischemic neuropathological damage emerges in both BCAO and MCAO models. Intraperitoneal injection of puerarin significantly helps improve all these brain ischemic disturbances.

Cloning of a rat cDNA encoding retinol dehydrogenase isozyme type III.

The primary and rate-limiting step in retinoic acid (RA) biosynthesis requires the conversion of retinol into retinal. Previously, two genes encoding retinol dehydrogenases (RoDH), which recognize holo-cellular retinol-binding protein as substrate, had been cloned, expressed and identified as members of the short-chain dehydrogenase/reductase (SDR) gene family. This work reports the cloning of a cDNA encoding a third RoDH isozyme, RoDH(III). The deduced amino-acid sequence of RoDH(III) indicates 97.8% identity with RoDH(I) and 82.3% identity with RoDH(II). RNase protection assays revealed RoDH(III) mRNA expression only in rat liver, in contrast to RoDH(I) and RoDH(II), which had their mRNA expressed in rat liver, kidney, lung, testis and brain. These data extend the insight that a subfamily of SDR isozymes, tissue-distinctively expressed, catalyzes the first step in RA biogenesis.

Cloning of a cDNA for a second retinol dehydrogenase type II. Expression of its mRNA relative to type I.

A retinol dehydrogenase, RoDH(1), which recognizes holo-cellular retinol-binding protein (CRBP) as substrate, has been cloned, expressed, and identified as a short-chain dehydrogenase/reductase (Chai, X., Boerman, M. H. E. M., Zhai, Y., and Napoli, J. L. (1995) J. Biol. Chem. 270, 3900-3904). This work reports the cloning and expression of a cDNA encoding a RoDH isozyme, RoDH(II). The predicted amino acid sequence verifies RoDH(II) as a short-chain dehydrogenase/reductase, 82% identical with RoDH(I). RoDH(II) recognized the physiological form of retinol as substrate, CRBP, with a Km of 2 mM. Similar to microsomal RoDH and RoDH(I), RoDH(II) had higher activity with NADP rather than NAD, was stimulated by ethanol and phosphatidyl choline, was not inhibited by the medium-chain alcohol dehydrogenase inhibitor 4-methylpyrazole, but was inhibited by phenylarsine oxide and the short-chain dehydrogenase/reductase inhibitor carbenoxolone. Northern blot analysis detected RoDH(I) and RoDH(II) mRNA only in rat liver, but RNase protection assays revealed RoDH(I) and RoHD(II) mRNA in kidney, lung, testis, and brain. These data indicate that short-chain dehydrogenases/reductase isozymes expressed tissue-distinctively catalyze the first step of retinoic acid biogenesis from the physiologically most abundant substrate, CRBP.

[Effect of puerarin on cerebral blood flow in dogs].

The hemodynamics of left intra- and extracranial arteries was determined by a pulsatile Doppler flowmeter and a transcranial Doppler system simultaneously in anesthetized dogs. Low doses of puerarin (50mg/kg, iv) did not alter the cerebral perfusion pressure, but reduced the flow velocities of both middle cerebral artery and anterior cerebral artery. The globle cerebral blood flow was enhanced due to dilatation of the intracranial arteries.

Efficacy of tablet huperzine-A on memory, cognition, and behavior in Alzheimer's disease.

AIM: To evaluate the efficacy and safety of tablet huperzine-A (Hup) in patients with Alzheimer's disease. METHODS: Using multicenter, prospective, double-blind, parallel, placebo controlled and randomized method, 50 patients were administered orally 0.2 mg (4 tablets) Hup and 53 patients were given po 4 tablets of placebo bid for 8 wk. All patients were evaluated with Wechsler memory scale, Hasegawa dementia scale, mini-mental state examination scale, activity of daily living scale, treatment emergency symptom scale, and measured with BP, HR, ECG, EEG, ALT, AKP, BUN, Cr, Hb, WBC, and urine routine. RESULTS: About 58% (29/50) of patients treated with Hup showed improvements in their memory (P < 0.01), cognitive (P < 0.01), and behavioral (P < 0.01 functions. The efficacy of Hup was better than placebo (36%, 19/53) (P < 0.05). No severe side effect was found. CONCLUSION: Hup is a promising drug for symptomatic treatment of Alzheimer's disease.

Cloning of a cDNA for liver microsomal retinol dehydrogenase. A tissue-specific, short-chain alcohol dehydrogenase.

Retinoic acid, a hormone biosynthesized from retinol, controls numerous biological systems by regulating eukaryotic gene expression from conception through death. This work reports the cloning and expression of a liver cDNA encoding a microsomal retinol dehydrogenase (RoDH), which catalyzes the primary and rate-limiting step in retinoic acid synthesis. The predicted amino acid sequence and biochemical data obtained from the recombinant enzyme verify it as a short-chain alcohol dehydrogenase. Like microsomal RoDH, the recombinant enzyme recognized as substrate retinol bound to cellular retinol-binding protein, had higher activity with NADP rather than NAD, was stimulated by ethanol or phosphatidylcholine, was not inhibited by 4-methylpyrazole, was inhibited by phenylarsine oxide and carbenoxolone and localized to microsomes. RoDH recognized the physiological form of retinol, holocellular retinol-binding protein, with a Km of 0.9 microM, a value lower than the approximately 5 microM concentration of holocellular retinol binding protein in liver. Northern and Western blot analyses revealed RoDH expression only in rat liver, despite enzymatic activity in liver, brain, kidney, lung, and testes. These data suggest that tissue-specific isozyme(s) of short chain alcohol dehydrogenases catalyze the first step in retinoic acid biogenesis and further strengthen the evidence that the "cassette" of retinol bound to cellular retinol-binding protein serves as a physiological substrate.

Clinical observation on treatment of bradyarrhythmia with Chinese herbal medicine.

This paper reports on treatment of 62 cases of bradyarrhythmia with Fu Ben Zeng Mai Tang, a decoction of herbal medicine which has the effect of warming and tonifying the heart and kidney, reinforcing qi and improving blood circulation. The result indicates that it is effective for quickening the heart rhythm, improving the symptoms and regulating the function of sinoatrial node. It is also good for regulating the function of endocrine and the immune and nervous system. This therapy was compared with Western medicine in the treatment of 24 cases.