Heterologous expression of a deacetylase and its application in L-glufosinate preparation.

With potent herbicidal activity, biocatalysis synthesis of L-glufosinate has drawn attention. In present research, NAP-Das2.3, a deacetylase capable of stereoselectively resolving N-acetyl-L-glufosinate to L-glufosinate mined from Arenimonas malthae, was heterologously expressed and characterized. In Escherichia coli, NAP-Das2.3 activity only reached 0.25 U/L due to the formation of inclusive bodies. Efficient soluble expression of NAP-Das2.3 was achieved in Pichia pastoris. In shake flask and 5 L bioreactor fermentation, NAP-Das2.3 activity by recombinant P. pastoris reached 107.39 U/L and 1287.52 U/L, respectively. The optimum temperature and pH for N-acetyl-glufosinate hydrolysis by NAP-Das2.3 were 45 °C and pH 8.0, respectively. The Km and Vmax of NAP-Das2.3 towards N-acetyl-glufosinate were 25.32 mM and 19.23 µmol mg-1 min-1, respectively. Within 90 min, 92.71% of L-enantiomer in 100 mM racemic N-acetyl-glufosinate was converted by NAP-Das2.3. L-glufosinate with high optical purity (e.e.P above 99.9%) was obtained. Therefore, the recombinant NAP-Das2.3 might be an alternative for L-glufosinate biosynthesis.

The Microbiota-Dependent Treatment of Wuzhuyu Decoction for Chronic Migraine Model Rat Associated with Anxiety-Depression Like Behavior.

We previously found that Wuzhuyu Decoction (WZYD) could affect central and peripheral 5-HT to relieve hyperalgesia in chronic migraine (CM) model rats, possibly related to gut microbiota. However, the exact role of gut microbiota has not been elucidated. Accumulating evidence points to the possibility of treating central nervous system disease via the gut-brain axis. In our study, the inflammatory soup-induced CM model rats presented depression- and anxiety-like behaviors which both related to insufficient 5-HT. It was found that antibiotic administration caused community dysbiosis, and proteobacteria became the main dominant bacteria. The bacteria related to short-chain fatty acids and 5-HT generation were reduced, resulting in reduced levels of 5-HT, tryptophan hydroxylase, and secondary bile acids. Functional prediction-revealed sphingolipid signaling pathway in CM rats was significantly decreased and elevated after WZYD treatment. The effect of WZYD could be weakened by antibiotics. The CM rats exhibited anxiety- and depression-like behavior with 5-HT and number of neurons decreased in the CA1 and CA2 regions of hippocampal. The treatment of WZYD could recover to varying degrees. Antibiotics combined with WZYD attenuate the effect of WZYD on increasing the 5-HT content and related protein expression in the brain stem, plasma and colon, reducing CGRP, c-Fos, and inflammatory factors. And antibiotics also led to colon length increasing and stool retention, so that the antimigraine effect was weakened compared with WZYD. This experiment revealed that gut microbiota mediated WZYD treatment of CM rats with anxiety-depression like behavior.

Wuzhuyu Decoction relieves hyperalgesia by regulating central and peripheral 5-HT in chronic migraine model rats.

BACKGROUND: Chronic migraine (CM) is a highly disabling and burdensome disease. Wuzhuyu decoction (WZYD), a clinical used formula to treat and prevent episodic migraine and CM, has been reported to relieve the hyperalgesia of CM and increase brainstem and blood serotonin (5-hydroxytryptamine, 5-HT) in migraine model rats in previous studies; yet the mechanism is unclear. PURPOSE: This study aimed to observe the hyperalgesia relief effect of WZYD and investigate the mechanistic association with the regulation on central and peripheral 5-HT. METHODS: WZYD with different doses (3.372, 1.686 and 0.843 g/kg∙d) and the positive drug - sumatriptan (5.83 mg/kg∙3 d) were intragastrically administered in inflammatory soup (IS)-induced CM model rats, respectively. Hyperalgesia was assessed by facial mechanical withdrawal threshold and tail-flick latency. 5-HT was determined by ELISA. Western blot analysis, immunohistochemistry and immunofluorescence determination, and 16S rRNA gene sequencing were performed. RESULTS: WZYD significantly relieved the hyperalgesia by elevating the facial mechanical withdrawal threshold and tail-flick latency. In WZYD groups, increased 5-HT and decreased calcitonin gene-related peptide in both the brainstem and plasma, downregulated TNF-α, IL-1ß, and c-fos expression in the brainstem were observed in dose-dependent manner. Interestingly, 5-HT in colon tissues were also observed, which is associated with upregulating tryptophan hydroxylase, serotonin transporter and Piezo1 expression and increasing 5-HT and chromogranin A in enterochromaffin cells. Disorder of the microbiota, function and metabolism was correlated with 5-HT synthesis. WZYD could regulate the abundance of Anaerostipes and Acidifaciens. CONCLUSION: WZYD has the pharmacological effect on relieving hyperalgesia in CM model rats, possibly by affecting central and peripheral 5-HT.

Evaluation on effect of new model of community-based hypertension self-management / 上海预防医学

To explore the new model of community-based hypertension self-management, through literature review and qualitative interviews with experts and stakeholders.Among 15 000 patients with informed consents and voluntary participation, 1 080 patients were randomly selected as the intervention group, and 588 patients were randomly selected from hypertension management database of community health service centers as the control group.Analysis of covariance based on rank scores was employed to compare the score changes in two survey questionnaires, before and after six months of the intervention project, and between the intervention group and the control group, on self-assessment of overall health status, and qual-ity of health care services received in the community.Analysis of covariance was employed to compare the changes of blood pressure. Results The self-assessment of overall health status significantly improved for the intervention group, compared with the control group (P<0.05).The blood pressure of the inter-vention group remained stable, while the diastolic pressure for the control group increased as compared with that before intervention, the difference was statistically significant (P<0.05).During six months of the intervention project, as to the incremental value of scores in dimensions like initiative and design of health care service system, the difference between the intervention group and the control group was statistically significant ( P <0 .05 ) . Conclusion The intervention based on the new model of community-based hypertension self-management has a positive role in improving the patients'overall health status, stabilizing the blood pressure, and optimizing hypertension health care services in community.

[Effect of TGF-ß1 expression on periodontal tissue reconstruction for rapid teeth movement through reducing resistance and distraction].

PURPOSE: To investigate the role of transforming growth factor-ß1 (TGF-ß1) in periodontal tissue reconstruction during rapid teeth movement through reducing resistance and distraction. METHODS: Twenty Beagle dogs were randomly divided into five groups according to time points: distraction for 5 days, 10 days, 15 days, retaining 10 days after distraction for 15 days, and retaining 90 days after distraction for 15 days. The mandibular first premolar was moved by using reducing resistance and distraction (experimental group) or conventional distraction (control group) randomly. For each group, tooth movement distance was measured and first premolar periodontal tissue pieces were obtained in scheduled time. Hematoxylin-eosin stain, Picric acid-Sirius red, and immunohistochemistry were performed. The data was analyzed by using SPSS 18.0 software package. RESULTS: The speed of tooth movement and periodontal reconstruction of the experimental group were significantly faster than the control group. TGF-ß1 positive expression in two groups had similar distribution area, and reached peak at most active stage of osteogenesis in both groups. But in different stages of distraction, the positive expression in the experimental group was significantly higher than the control group(P<0.05). CONCLUSIONS: Compared with conventional orthodontic method, reducing resistance and distraction can obviously increase the expression of TGF-ß1 of tension side of the distracted tooth, and accelerate periodontal tissue reconstruction.

Identification of a retroelement from the resurrection plant Boea hygrometrica that confers osmotic and alkaline tolerance in Arabidopsis thaliana.

Functional genomic elements, including transposable elements, small RNAs and non-coding RNAs, are involved in regulation of gene expression in response to plant stress. To identify genomic elements that regulate dehydration and alkaline tolerance in Boea hygrometrica, a resurrection plant that inhabits drought and alkaline Karst areas, a genomic DNA library from B. hygrometrica was constructed and subsequently transformed into Arabidopsis using binary bacterial artificial chromosome (BIBAC) vectors. Transgenic lines were screened under osmotic and alkaline conditions, leading to the identification of Clone L1-4 that conferred osmotic and alkaline tolerance. Sequence analyses revealed that L1-4 contained a 49-kb retroelement fragment from B. hygrometrica, of which only a truncated sequence was present in L1-4 transgenic Arabidopsis plants. Additional subcloning revealed that activity resided in a 2-kb sequence, designated Osmotic and Alkaline Resistance 1 (OAR1). In addition, transgenic Arabidopsis lines carrying an OAR1-homologue also showed similar stress tolerance phenotypes. Physiological and molecular analyses demonstrated that OAR1-transgenic plants exhibited improved photochemical efficiency and membrane integrity and biomarker gene expression under both osmotic and alkaline stresses. Short transcripts that originated from OAR1 were increased under stress conditions in both B. hygrometrica and Arabidopsis carrying OAR1. The relative copy number of OAR1 was stable in transgenic Arabidopsis under stress but increased in B. hygrometrica. Taken together, our results indicated a potential role of OAR1 element in plant tolerance to osmotic and alkaline stresses, and verified the feasibility of the BIBAC transformation technique to identify functional genomic elements from physiological model species.

Maize membrane-bound transcription factor Zmbzip17 is a key regulator in the cross-talk of ER quality control and ABA signaling.

Abiotic stresses disrupt protein folding and induce endoplasmic reticulum (ER) stress, which in turn activates the unfolded protein response (UPR) to aid in the refolding or degradation of misfolded proteins. The phytohormone ABA regulates many aspects of plant development and plays a central role in the stress response; however, the role of ABA in transducing stress signals to activate the UPR has not been recognized. In this study, a gene encoding the maize ortholog of AtbZIP17, a transmembrane transcription factor functioning as an ER stress transducer, was identified from the MaizeGDB database, and designated ZmbZIP17. ZmbZIP17 was induced by both ABA and ER stress-eliciting agents such as dithiotreitol (DTT) and tunicamycin (TM). Transiently expressed yellow fluorescent protein (YFP)-ZmbZIP17 co-localized with the ER marker HDEL-mCherry under control conditions, but partially translocated into the nucleus upon TM treatment or removal of the transmembrane domain. TM-induced processing of ZmbZIP17 was confirmed by Western blot analysis. When overexpressed in Arabidopsis, ZmbZIP17 triggered ER stress response gene expression and tolerance to DTT and TM, elevated ABA-responsive gene expression and ABA sensitivity both pre- and post-germination. Additionally, ABA was found to induce ER stress response gene expression, alone or synergistically with ZmbZIP17, in the absence of DTT or TM; while ZmbZIP17 was capable of interacting with ABA-responsive cis-elements (ABREs) that exist in promoters of known ABA-responsive genes. Together, our results reveal a direct link between ER stress and ABA signaling pathways involving the ZmbZIP17 transcription factor.

Predictive value of serum uric acid on cardiovascular disease and all-cause mortality in urban Chinese patients.

BACKGROUND: The association between increased serum uric acid (SUA) levels and cardiovascular risk has been debated for decades. Several large studies have provided conflicting results regarding the clinical significance of elevated SUA levels in cardiovascular disease (CVD) or cerebrovascular disease. The aim of this study was to investigate the relationship between SUA and CVD and all-cause mortality and their potential diagnostic value. METHODS: A total of 3570 in-patients ranging in age from 56 to 95 years (mean (67.36 +/- 11.36) years) were selected from 20 hospitals in Beijing and Shanghai. A carefully designed questionnaire was used to gather baseline data of each patient. All patients were divided into two main groups according to their SUA levels: high SUA and normal SUA groups. Serum indices and other important parameters were measured. RESULTS: Compared with normal SUA group, high SUA group had significant difference in systolic blood pressure (SBP), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), body mass index (BMI), and age (P < 0.05 or P < 0.01). High SUA prevailed in female and patients with history of essential hypertension, while history of smoking and diabetes showed no significant difference between two groups. All-cause and CVD mortality occurred more frequently in high SUA group than in normal SUA group. In the accumulative survival analysis, high SUA group had lower survival rate than normal SUA group both in CVD and all-cause mortality. COX regression analysis indicated that the history of smoking, age and high SUA were independent risk factors for the development of CVD. CONCLUSIONS: These preliminary observations suggest that patients with high SUA levels would face higher risk of mortality. SUA measurement may be applied as a routine predictor for clinical assessment.

[Multi-factor model of quality of life in patients with acute coronary syndrome].

OBJECTIVE: To detect the risk factors of quality of life (QOL) in patients with acute coronary syndrome (ACS), so as to help doctors to recognize the risk population with impaired QOL. METHODS: 403 hospitalized ACS patients, 252 males and 151 females, aged 67 12, 168 with ST-segment elevation myocardial infarction (STEMI), 4 with non-ST-segment elevation myocardial infarction (NSTEMI), and 74 with unstable angina pectoris (UAP), were surveyed with short-form-36 (SF-36) scale (Chinese version), including physical component summary (PCS) and mental component summary (MCS) so as to analyze the QOL, hospital anxiety depression scales (HADS) so as to detect the anxiety status. Multiple linear regression analysis was conducted to identify the variables associated with QOL. RESULTS: Of the 403 patients 84 (20.8%) suffered from anxiety, 43 (10.7%) suffered from depression, and 144 (35.7%) suffered from depression combined with anxiety symptoms . Age (r = -0.237, P < 0.001), being female (r = -0.183, P < 0.001), depression (r = -0.180; P < 0.001), and anxiety (r = -0.211, P <0. 001) were independent risk factors of physical QOL. Age (r = -0.117, P = 0.01), depression (r =-0.169, P = 0.004), and anxiety (r = -0.215, P < 0.001) were independently negatively correlated with mental QOL. Smoking was unexpectedly a protective factor for MCS (r = 0.157, P = 0.001). CONCLUSION: Depression and anxiety are common in patients diagnosed as with ACS, and appear to be related to significant worsening of QOL. The patients being female, with elder age, and suffering from depression and/or anxiety may be with impaired QOL, and need more care and effective intervention.

[The relationship between ankle brachial index and all-cause and cardiovascular disease mortality in Chinese metabolic syndrome male cohort].

OBJECTIVE: Ankle brachial index (ABI) is thought to be an efficient means of objectively assessing the potency of lower extremity arterial system. This study is aimed to evaluate the relationship between ABI and cardiovascular disease CVD mortality in Chinese male patients with metabolic syndrome (MS). METHODS: 1224 Chinese male patients with MS were selected from Beijing and Shanghai and the baseline examinations were carried out. All the participants were divided into 2 main groups: ABI < or = 0.9 (n = 268) and ABI 0.9 - 1.4 (n = 956). and they were followed up for (13.2 +/- 2.7) months. RESULTS: As to baseline characteristics, age, systolic blood pressure (SBP), hypertensive disease and diabetes mellitus morbidity and smoking history had significant difference between the 2 groups (P < 0.05 or P < 0.01). All-cause mortality and CVD mortality decreased gradually while the ABI increased from 0.4 to 1.4. With Cox regression analysis, relative ratio (RR) value of all-cause mortality and CVD mortality also showed the decreasing trend with the rising of ABI. CONCLUSION: In Chinese male patients with MS, ABI is one of the most important parameters in indicating possible prognosis and foreseeing all-cause and CVD mortality. Male subjects with relatively old age, higher systolic blood pressure, hypertensive disease and diabetes mellitus morbidity, smoking history may be associated with lower ABI (< or = 0.9) and relatively higher all-cause and CVD mortality. Our results suggest the urgent need for frequent measurement of the ABI in clinical practice before diagnosing peripheral artery disease and making therapeutic decision, especially in some high-risk population such as male patients with MS.

[The relation between post-acute coronary syndrome anxiety and serum levels of inflammatory factors].

OBJECTIVE: To explore the role of inflammatory factors (interleukin-6, tumor necrosis factor alpha, high sensitive C reactive protein) in the pathogenesis of anxiety after acute coronary syndrome (ACS) by investigating the serum levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha) and high sensitive C reactive protein (hsCRP). METHODS: Serum IL-6, TNFalpha and hsCRP levels were measured in 105 ACS patients within 7 days after onset of the event. Symptoms of anxiety were evaluated by self-reporting standardized questionnaire, using a validated Chinese version of Hospital Anxiety and Depression Scale (HADS-A)-Anxiety Subscale (7 items) within 72 hours of the event. Demographic and clinical data including cardiac risk factors were recorded. RESULTS: The median of TNFalpha was lower in the anxious patients with ACS than that of non-anxious ones (44.55 vs 61.70, P = 0.004). Neither the levels of hsCRP nor the levels of IL-6 were found to be different between the groups with anxiety and without. CONCLUSION: Anxiety after ACS does not increase the inflammatory reaction in the ACS patients.