[Dynamic control of ERG20 expression to improve production of monoterpenes by engineering Saccharomyces cerevisiae].

Monoterpenes are widely used in cosmetics, food, medicine, agriculture and other fields. With the development of synthetic biology, it is considered as a potential way to create microbial cell factories to produce monoterpenes. Engineering Saccharomyces cerevisiae to produce monoterpenes has been a research hotspot in synthetic biology. In S. cerevisiae, the production of geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) is catalyzed by a bifunctional enzyme farnesyl pyrophosphate synthetase(encoded by ERG20 gene) which is inclined to synthesize FPP essential for yeast growth. Therefore, reasonable control of FPP synthesis is the basis for efficient monoterpene synthesis in yeast cell factories. In order to achieve dynamic control from GPP to FPP biosynthesis in S. cerevisiae, we obtained a novel chassis strain HP001-pERG1-ERG20 by replacing the ERG20 promoter of the chassis strain HP001 with the promoter of cyclosqualene cyclase(ERG1) gene. Further, we reconstructed the metabolic pathway by using GPP and neryl diphosphate(NPP), cis-GPP as substrates in HP001-pERG1-ERG20. The yield of GPP-derived linalool increased by 42.5% to 7.6 mg·L~(-1), and that of NPP-derived nerol increased by 1 436.4% to 8.3 mg·L~(-1). This study provides a basis for the production of monoterpenes by microbial fermentation.

Effect of wrist-ankle acupuncture therapy combined with auricular acupuncture on cancer pain: A four-parallel arm randomized controlled trial.

BACKGROUND: Cancer pain affects the quality of life of cancer patients; therefore, various methods exist for alleviating the adverse effects caused by cancer pain. Nonpharmacological intervention is regarded as an important means of auxiliary therapy for drug treatment, with acupuncture receiving the most attention; However, there are numerous types of acupuncture therapies, including acupuncture, wrist-ankle acupuncture (WAA) and auricular acupuncture (AA). Previous studies have demonstrated that all types of acupuncture therapy can alleviate cancer pain. However, the effects and pathways of different acupuncture treatments are not similar, and it is unknown whether single therapy or combination therapy has better analgesic effects. This study aimed to examine the effect of WAA therapy combined with AA on cancer pain. DESIGN: A randomized controlled trial. METHOD: A total of 160 patients were selected and randomly divided into groups A, B, C and D, with 40 patients in each group. Group A received conventional analgesia alone, with opioids administered based on the World Health Organization (WHO) 3-tiered "cancer pain ladder". Group B received WAA, in addition to the treatment received by group A. Group C received AA, in addition to the treatment received by group A. Group D received WAA combined with AA, in addition to the treatment received by group A. Analgesic effects and analgesic drug use before and 3, 5 and 7 days after treatment were observed in each group. RESULT: A total of 159 patients were included in the analysis. The verbal rating scale (VRS) and numeric rating scale (NRS) scores for patients who received mono-acupuncture therapy and combination therapy for 1 week were significantly different from those of the control group. Combination therapy had a stronger effect on the VRS score and a faster onset time, based on the NRS score, and the patients who received combination therapy had reduced analgesic drug use. CONCLUSION: WAA combined with AA can more quickly reduce pain symptoms with more lasting analgesic effects and can effectively reduce analgesic drug use.

[Pharmacological effect of Schisandrae Chinensis Fructus and relative active components on cardiovascular and cerebrovascular diseases].

Cardiovascular and cerebrovascular diseases (CCDs) are the primary causes of death in Chinese adults. With the increase in morbidity and mortality rates and the decrease in the age of onset, CCD becomes a very natural target for traditional Chinese medicine. Schisandrae Chinensis Fructus (SCF) is the dry ripe fruit of Schisandra chinensis (Turcz.) Baill., which features a sweet and sour taste and the effects of calming the heart and tranquilizing the mind. It is mainly used for treatment of dysphoria and palpitation, insomnia and dreamful sleep due to the lack of spirit preservation. The main components of SC include lignans, volatile oils and polysaccharides. This review summarized the pharmacological effects of SC and its active components in the treatment of CCDs. The results showed that SCF and its active components protect against cardiovascular diseases mainly through the antioxidant, apoptosis inhibition and anti-inflammatory mechanisms. In addition, they protect against cerebrovascular diseases mainly by increasing energy metabolism, regulating autophagy and inhibiting apoptosis, antioxidant, and regulating nerve neurotransmitters and circadian genes. In conclusion, lignans are the most active components in SCF. This study provides a reference for the clinical research and utilization of SCF, as well as the application basis for co-treatment of cardiovascular and cerebrovascular diseases.

Tongxinluo Inhibits Renal Fibrosis in Diabetic Nephropathy: Involvement of the Suppression of Intercellular Transfer of TGF-[Formula: see text]1-Containing Exosomes from GECs to GMCs.

Glomerular mesangial cells (GMCs) activation is implicated in the pathogenesis of diabetic nephropathy (DN). Our previous study revealed that high glucose (HG)-treated glomerular endothelial cells (GECs) produce an increased number of TGF-[Formula: see text]1-containing exosomes to activate GMCs through the TGF-[Formula: see text]1/Smad3 signaling pathway. We also identified that Tongxinluo (TXL), a traditional Chinese medicine, has beneficial effects on the treatment of DN in DN patients and type 2 diabetic mice. However, it remained elusive whether TXL could ameliorate renal structure and function through suppression of intercellular transfer of TGF-[Formula: see text]1-containing exosomes from GECs to GMCs. In this study, we demonstrate that TXL can inhibit the secretion of TGF-[Formula: see text]1-containing exosomes from HG-treated GECs. Furthermore, exosomes produced by HG induced-GECs treated with TXL cannot trigger GMC activation, proliferation and extracellular matrix (ECM) overproduction both in vitro and in vivo. These results suggest that TXL can prevent the transfer of TGF-[Formula: see text]1 from GECs to GMCs via exosomes, which may be one of the mechanisms of TXL in the treatment of DN.

Effects of high hydrostatic pressure and thermal processing on bioactive compounds, antioxidant activity, and volatile profile of mulberry juice.

The aim of this study was to investigate the effects of high hydrostatic pressure and thermal processing on microbiological quality, bioactive compounds, antioxidant activity, and volatile profile of mulberry juice. High hydrostatic pressure processing at 500 MPa for 10 min reduced the total viable count from 4.38 log cfu/ml to nondetectable level and completely inactivated yeasts and molds in raw mulberry juice, ensuring the microbiological safety as thermal processing at 85 ℃ for 15 min. High hydrostatic pressure processing maintained significantly (p < 0.05) higher contents of total phenolic, total flavonoid and resveratrol, and antioxidant activity of mulberry juice than thermal processing. The main volatile compounds of mulberry juice were aldehydes, alcohols, and ketones. High hydrostatic pressure processing enhanced the volatile compound concentrations of mulberry juice while thermal processing reduced them in comparison with the control. These results suggested that high hydrostatic pressure processing could be an alternative to conventional thermal processing for production of high-quality mulberry juice.

Antioxidation and anti-inflammatory activity of Tang Bi Kang in rats with diabetic peripheral neuropathy.

BACKGROUND: Tang Bi Kang (TBK) is a traditional Chinese medicine granule. It has been shown to have effects on nerve conduction velocity deficits, blood-related factors and oxidative stress. This study was undertaken to evaluate proposed antioxidative and anti-inflammatory activity of Tang Bi Kang in rats with diabetic peripheral neuropathy (DPN). METHODS: DPN was induced in male Wistar rats by intraperitoneal administration of streptozocin (STZ) (60 mg/kg.b.w) for 8 weeks. Fasting blood glucose (FBG) levels were measured in the blood obtained by clipping the tails of the rats. Tail-flick tests were conducted with a tail-flick analgesic meter. Motor and sensory nerve conduction velocities (MNCV and SNCV) of sciatic nerve were measured directly at two sites using a Functional Experiment System. Oxidative stress makers such as malondialdehyde (MDA), superoxide-dismutase (SOD) and glutathione peroxidase (GSH-Px), inflammatory cytokines such as interleukin (IL)-6, and tumour necrosis factor (TNF)-α were estimated. The statistical analysis of results was carried out using Student t-test and one-way analysis of variance (ANOVA), followed by least-significant difference post hoc with SPSS. RESULTS: The administration of TBK for 4 weeks in DPN rats resulted in a significant decrease in FBG levels compared to untreated DPN rats. There was a significant increase in MNCV and SNCV in the DPN rats compared to untreated DPN rats. Serum level of MDA was significantly reduced while the activities of SOD and GSH-pX were significantly increased in the TBK treated DPN rats. TBK prevented DPN-induced increase in the serum levels of IL-6 and TNF-α. CONCLUSION: The results of this study demonstrate that the therapeutic effect of TBK on DPN rats may be associated with the antioxidative and anti-inflammatory responses.

Effects of the combined extracts of Herba Epimedii and Fructus Ligustri Lucidi on bone mineral content and bone turnover in osteoporotic rats.

BACKGROUND: The decoction combination of Herba Epimedii and Fructus Ligustri Lucidi has been used to treat osteoporosis for almost 50 years by practitioners of traditional Chinese medicine. However, it is unclear what specific effects this combination of herbs has on the skeleton. The aim of this study was to assess the effects of the combined extracts from Herba Epimedii and Fructus Ligustri Lucidi on the bone turnover and bone mineral content in a rat model of osteoporosis induced by retinoic acid. METHODS: Fifty male Wistar rats were randomly assigned to the normal control group, osteoporosis model group, or treatment groups in which osteoporosis was induced and then the combined extracts of Herba Epimedii and Fructus Ligustri Lucidi were administered at 50, 100, or 200 mg/kg/day for 3 weeks via oral gavage. The rat osteoporosis model was induced by intragastric administration of 70 mg/kg/day of retinoic acid for 2 weeks. Bone turnover markers, bone biomechanical properties, and the calcium and phosphorus content of the right tibia and serum were measured. RESULTS: The retinoic acid administration decreased the bone mass and the contents of calcium and phosphorus in the bone mineral, weakened the biomechanical properties, and increased bone turnover by stimulating bone resorption and collagen metabolism. Treatment with the combined extracts of Herba Epimedii and Fructus Ligustri Lucidi significantly mitigated the effects of osteoporosis on the rats by decreasing bone metabolism, improving the bone mineral content, and increasing the biomechanical properties. CONCLUSIONS: The results of this study highlight the anti-osteoporosis effects of the combined extracts of Herba Epimedii and Fructus Ligustri Lucidi. These findings may contribute to the development of natural anti-osteoporosis herbal medicines.

Tongxinluo ameliorates renal structure and function by regulating miR-21-induced epithelial-to-mesenchymal transition in diabetic nephropathy.

Diabetic nephropathy (DN) is one of the most important diabetic microangiopathies. The epithelial-to-mesenchymal transition (EMT) plays an important role in DN. The physiological role of microRNA-21 (miR-21) was closely linked to EMT. However, it remained elusive whether tongxinluo (TXL) ameliorated renal structure and function by regulating miR-21-induced EMT in DN. This study aimed to determine the effect of TXL on miR-21-induced renal tubular EMT and to explore the relationship between miR-21 and TGF-ß1/smads signals. Real-time RT-PCR, cell transfection, in situ hybridization (ISH), and laser confocal microscopy were used, respectively. Here, we revealed that TXL dose dependently lowered miR-21 expression in tissue, serum, and cells. Overexpression of miR-21 can enhance α-smooth muscle actin (SMA) expression and decrease E-cadherin expression by upregulating smad3/p-smad3 expression and downregulating smad7 expression. Interestingly, TXL also increased E-cadherin expression and decreased α-SMA expression by regulating miR-21 expression. More importantly, TXL decreased collagen IV, fibronectin, glomerular basement membrane, glomerular area, and the albumin/creatinine ratio, whereas it increased the creatinine clearance ratio. The results demonstrated that TXL ameliorated renal structure and function by regulating miR-21-induced EMT, which was one of the mechanisms to protect against DN, and that miR-21 may be one of the therapeutic targets for TXL in DN.

[Effects of a glucocorticoid on development of kidney deficiency syndrome in a rat model of asthma].

OBJECTIVE: To observe the effects of dexamethasone (DXM) on changes of syndrome and hypothalamic-pituitary-adrenal (HPA) axis of asthmatic rats at different stages of intervention (before withdrawal, during withdrawal and after withdrawal). METHODS: Asthma was induced in rats by inhalation of 1% ovalbumin. Rats in DXM group were treated with different concentrations of DXM by intra-abdominal injection. At the different stages of DXM treatment (before withdrawal, during withdrawal and after withdrawal), symptoms of the rats were observed to differentiate the syndromes according to yin-deficiency and yang-deficiency measurement standard. Content of serum corticosterone (CORT) was detected by enzyme-linked immunosorbent assay, and contents of plasma adrenocorticotropic hormone (ACTH) and bone Gla-protein (BGP), and content of corticotropin-releasing hormone (CRH) in hypothalamic homogenate were detected by radioimmunoassay. RESULTS: Rats in the DXM group were differentiated as yin-deficiency syndrome before the stage of hormone withdrawal. During the period of hormone withdrawal and after the withdrawal, syndromes of rats in the DXM group accorded with the standards of yin- and yang-deficiency. During the period of treatment with DXM, contents of CORT, ACTH and CRH were significantly decreased, but recovered after DXM withdrawal. BGP level in the DXM group was lower than those in the normal group and the model group during all the stages. CONCLUSION: Asthmatic rats treated with DXM have different syndrome characteristics in different stages. Kidney deficiency will be caused by DXM treatment.

Triterpenoid saponins with anti-inflammatory activity from Codonopsis lanceolata.

Six triterpenoid saponins, including a new compound named codonolaside III, were isolated from the roots of Codonopsis lanceolata. The spectral and chemical data revealed the structure of codonolaside III to be 3- O-[ beta- D-xylopyranosyl -(1-->3)- beta- D-glucuronopyranosyl]-3 beta,16 alpha-dihydroxyolean-12-ene-28-oic acid 28- O-[ beta- D-xylopyranosyl-(1-->4)- alpha- L-rhamnpyranosyl-(1-->2)][ beta- D-glucopyranosyl -(1-->4)]- alpha- L-arabinopyranosyl ester. The xylene-induced mouse ear edema inhibitory effect assay disclosed codonolaside and codonolasides I - III as the major anti-inflammatory constituents in this crude drug.

Synergistic effects of atenolol and amlodipine for lowering and stabilizing blood pressure in 2K1C renovascular hypertensive rats.

AIM: To test the synergistic effects of atenolol and amlodipine on lowering blood pressure (BP) and reducing blood pressure variability (BPV) in 2-kidney, one-clip (2K1C) renovascular hypertensive rats. METHODS: Forty-eight 2K1C renovascular hypertensive rats were randomly divided into 6 groups. They were respectively given 0.8% carboxymethylcellulose sodium (control), atenolol (10.0 mg/kg), amlodipine (1.0 mg/kg), and combined atenolol and amlodipine (low dose: 5.0+0.5 mg/kg; intermediate dose: 10.0+1.0 mg/kg; high dose: 20.0+2.0 mg/kg). The drugs were given via a catheter in a gastric fistula. BP was recorded for 25 h from 1 h before drug administration to 24 h after administration. RESULTS: Compared with BP before medication, all 3 doses of combined atenolol and amlodipine significantly decreased the BP at 24 h after administration, except for the low dose on diastolic BP. Compared with the control group, all 3 doses of combined atenolol and amlodipine significantly reduced the average BP levels for the 24 h period after administration; furthermore, the high and intermediate doses also significantly decreased the BPV levels for the same period. The q values calculated by probability sum analysis for systolic and diastolic BP for the 24 h period after administration were 2.29 and 1.45, respectively, and for systolic and diastolic BPV for the same period they were 1.41 and 1.60, respectively. CONCLUSION: There is significant synergism between atenolol and amlodipine in lowering and stabilizing BP in 2K1C renovascular hypertensive rats.

Two useful methods for evaluating antihypertensive drugs in conscious freely moving rats.

AIM: Computerized analysis of blood pressure in conscious freely moving rats is a sound technique for physiological and pharmacological studies. The present work, based on this technique, was designed to introduce two useful methods for the evaluation of antihypertensive drugs in conscious spontaneously hypertensive rat (SHR). They were the directly intragastric administration of drugs and modified probability sum test for evaluating the synergism of the combination of two drugs. METHODS AND RESULTS: (1) Directly intragastric administration was used in conscious rats. A catheter was inserted into stomach immediately after arterial catheter insertion. Three days after operation, blood pressure was recorded and drug might be given intragastrically via the gastric catheter. (2) Modified probability sum test was used to evaluate the synergism of two drugs. The formula was: q=P(A+B)/(PA+PB-PAxB). With this method, it was obtained: q=1.32 for the effects of the combination of atenolol and nitrendipine (20 mg/kg+10 mg/kg) on systolic blood pressure; q=1.41 for the effects of the combination of atenolol and amlodipine (10 mg/kg+1 mg/kg) on systolic blood pressure. CONCLUSION: The two methods introduced by the present work will be important and useful for antihypertensive drug evaluation in conscious freely moving rats.