To establish induction and liquid culture system for hairy roots of Danshen (Salvia miltiorrhiza), Agrobacterium rhizogenes A4, LBA9402, 15834 as test bacterium were used to infect aseptic leaves of Danshen. The hairy roots were induced and positive transgenic hairy roots were selected with PCR using rolB and rolC as the target gene. Then hairy roots of S. miltiorrhiza were harvested and salvianolic acids were extracted with 70% methanol containing 1% formic acid. The content of salvianolic acid B (SalB) and rosmarinic acid (RA) were determined by HPLC. According to the above research results, the Danshen hairy roots induced by A. rhizogenes LBA9402 were inoculated into the following group of culture media: MSOH, MS, B5, and 6,7-V liquid media. Then the same methods of extraction and determination for the content of Danshen hairy roots were adopted. Last, the hairy roots of S. miltiorrhiza induced by A. rhizogenes LBA9402 were inoculated into the MSOH liquid media with different pH values. The content of salvianolic acid were extracted with 70% methanol containing 1% formic acid and determined by HPLC. As a result, three kinds of A. rhizogenes A4, LBA9402, 15834 could induce hairy roots and Ri plasmids were integrated into the genome of S. miltiorrhiza by PCR. Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid, which were (3.27 ± 0.37)% [including (1.04 ±0.36)% of RA and (2.22 ± 0.29)% of SalB] and (3.17 ± 0.20)% [including (0.92 ± 0.31)% of RA and (2.25 ± 0.26)% of SalB], respectively. Hairy roots induced by A. rhizogenes LBA9402 when they were cultured in MSOH liquid media produced much more salvianolic acid, which was (4.56 ± 0.36)%, including (1.12 ± 0.26)% of RA and (3.44 ± 0.23)% of SalB. Hairy roots induced by A. rhizogenes LBA9402 produced the most salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81, which was 4.85%, including 1.16% of RA and 3.69% of SalB. So Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81. The research had established the foundation on genetic engineering to improve the quality of S. miltiorrhiza.
Agrobacterium ; physiology ; Benzofurans ; analysis ; metabolism ; Cell Culture Techniques ; instrumentation ; methods ; Cinnamates ; analysis ; metabolism ; Culture Media ; chemistry ; metabolism ; Depsides ; analysis ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Salvia miltiorrhiza ; chemistry ; growth & development ; metabolism ; microbiology

yin-yang deficiency,and there were significant difference between different syndromes P

OBJECTIVE: To observe the therapeutic effect of probucol on serum malondialdehyde (MDA) and superoxide dismutase (SOD) in patients with primary hypertension. METHODS: A randomized study was performed on 40 patients with hypertension. The patients were randomly assigned to the control (levamlodipine besylate 2.5 mg/d plus benazepril 10 mg/d, n=20) or probucol group (levamlodipine besylate 2.5 mg/d plus benazepril 10 mg/d plus probucol 500 mg/d, n=20). An additional twenty healthy people were enrolled in the study (normal group). All subjects were followed up for a period of four weeks. Lipids and hepatic/renal function were measured at baseline and after 4 weeks. The levels of serum MDA and SOD activity were assayed by chemical colorimetry, and other indices, including blood pressure, lipids and hepatic/renal function, were measured at baseline and after 4 weeks. RESULTS: Compared to the normal group, the levels of MDA in all of the hypertension patient groups were higher, SOD was lower. The antihypertensive treatment decreased serum MDA levels but increased SOD content, and probucol treatment exaggerated these effects, with greater reduction of serum MDA levels and greater increase of SOD content. CONCLUSION The treatment with probucol can improve oxidative stress in hypertension patients, resulting in reduced serum MDA levels and improved SOD activity, thus contributing agreater antihypertensive effect.
Adult ; Antioxidants ; therapeutic use ; Female ; Humans ; Hypertension ; drug therapy ; physiopathology ; Male ; Malondialdehyde ; blood ; Middle Aged ; Oxidative Stress ; drug effects ; Probucol ; therapeutic use ; Superoxide Dismutase ; blood

italic>Salvia miltiorrhiza Bunge is a traditional Chinese medicinal herb widely used to treat cardiovascular and cerebrovascular diseases at clinic. Its main water-soluble components are rosmarinic acid (RA) and salvianolic acid B (SAB), which are produced by phenylpropanoid pathway. 4-Hydroxyphenylpyruvate reductase (HPPR) is a key enzyme in phenylpropanoid metabolism pathway. SmHPPR1 was cloned from S. miltiorrhiza and was constructed into plant expression vector pJR-SmHPPR1. On this basis, SmHPPR1 transgenic Arabidopsis plants were induced and the content of 4-hydroxyphenyllactic acid (pHPL) was determined. SmHPPR1-overexpressing (SmHPPR1-OE) hairy roots of S. miltiorrhiza were obtained and the concentration of active components and transcriptome analysis were performed. The results showed that the concentration of pHPL in SmHPPR1 transgenic Arabidopsis T1 was 0.594 mg·g^-1 dry weight. The concentration of RA, SAB and total salvianolic acid in SmHPPR1-OE-3 hairy roots were 1.09, 1.29, 1.15 times of that in control-3, respectively, and the content of Danshensu was 36.26% of that in control-3. Transcriptomic analysis revealed that overexpression of SmHPPR1 caused the upregulation of other phenylpropanoid pathway genes like SmTAT2. Protein-protein interaction indicated CYT (TR74706_c0_g1), NADP⁺ (TR26565_c0_g1) and NADP⁺ (TR68771_c0_g1) is the central node of the network and participated in metabolic process and cellular process. The tracking work in this study proved that SmHPPR1 could catalyze the reduction of 4-hydroxyphenylpyruvic acid to 4-hydroxyphenyllactic acid in SmHPPR1 transgenic Arabidopsis, and SmHPPR1-overexpressing in hairy roots of S. miltiorrhiza could increase the concentration of salvianolic acids through synergistically regulating other pathway genes.

ObjectiveTo observe the effect ofZuogui Jiangtang Jieyu Formula on the expressions of glutamate, NR2A and NR2B in hippocampus of diabetic rats with depression;To explore the mechanism of protective effect. Methods Diabetes with depression rat models were established and then were randomly divided into the model group, positive medicine group, high-, medium- and low-doseZuogui Jiangtang Jieyu Formula groups. Normal rats acted as normal group, 16 rats per group. After 28 days of administration, Open-field test was used to detect the behavior of the rats;glutamate content of hippocampus was detected by ELISA;the expressions of NR2A and NR2B in rat hippocampus were detected by immunofluorescence.Results Compared with normal group, automatic activity times of rats in model group decreased significantly (P<0.01);both glutamate content (P<0.01) and expressions of NR2A, NR2B (P<0.01) increased significantly. Compared with the model group, automatic activity times of rats in positive medicine group and high-doseZuogui Jiangtang Jieyu Formula group significantly increased (P<0.01);glutamate content dropped (P<0.01);expressions of NR2A and NR2B decreased (P<0.05).ConclusionZuogui Jiangtang Jieyu Formula can improve depressive behavior of diabetic rats with depression, which may be related to the regulation of glutamate content and expressions of NR2A and NR2B in hippocampus.

Objective:To explore the intervention effect of motivational interviews based on timing theory on self-efficacy, negative affect and coping styles of parents with infantile spasms children.Methods:Cluster sampling was used to select 82 parents of infantile spasms hospitalized in the Department of Neurology of a children’s hospital, a three-A hospital from January 2019 to October 2019. They were divided into control group and observation group with 41 cases each according to random number table. The control group received routine health education, and the observation group received five motivational interviews based on timing theory interventions on the basis of routine care. The effect of the intervention was evaluated by General Self-Efficacy Scale (GSES), Hospital Anxiety and Depression Scale (HADS), and the Chinese version of Coping Health Inventory for Parents (CHIP) before intervention, on the day of discharge, and 3 months after discharge.Results:Before the intervention, there was no significant difference in the scores of GSES, HADS and CHIP scales between the parents of the two groups ( P>0.05). After intervention, The GSES scores of the observation group on the day of discharge and 3 months after discharge were (19.63±0.87) and (22.58±1.28) points, which were significantly higher than (18.92±0.74) and (19.46±1.25) points of the control group. The difference between both groups was statistically significant ( t values were -3.865, -10.926, P<0.01). HADS-A/HADS-D scores of the observation group on the day of discharge and 3 months after discharge were (12.50±0.82), (10.50±0.87) and (9.78±0.80), (8.63±0.87) points, respectively. The HADS-A/HADS-D scores of the control group on the day of discharge and 3 months after discharge were (12.92±0.74), (11.72±0.99) and (10.23±0.78), (9.38±1.04) points, respectively. The difference was statistically significant ( t values were 2.412-5.764, P<0.05 or 0.01). The observation group scores on CHIP subscales on the day of discharge and 3 months after discharge are higher than the control group, the difference was statistically significant (t values were -7.93--2.490, P<0.05 or 0.01). Conclusions:Motivational interviews based on timing theory can enhance parents’ self-efficacy, improve their negative emotions and family coping styles, and thereby promote the recovery of children.

OBJECTIVETo evaluate the effects of policosanol on serum lipid and heme oxygenase-1 (HO-1) in patients with hyperlipidemia.

METHODSThis randomized, open study included 72 patients with hyperlipidemia. The patients were randomly assigned to treatment group (policosanol, 20 mg/d, n = 36) or placebo group (placebo, two tablets/d, n = 36). The levels of serum lipids, hypersensitive C-reactive protein (hs-CRP), HO-1 were assessed before and after 16 weeks treatment. Drug-induced adverse effects and events were recorded during the observation period. The serum HO-1 was measured by ELISA.

RESULTS(1) After 16 weeks, all parameters remained unchanged in the placebo group; the level of TC decreased from (7.01 ± 1.03) mmol/L to (5.66 ± 0.83) mmol/L (-19.4%, P < 0.01), the level of LDL-C decreased from (4.78 ± 0.72) mmol/L to (3.70 ± 0.69) mmol/L (-22.5%, P < 0.01) in the treatment group. TG and HDL-C levels remained unchanged (P > 0.05) while the level of HO-1 significantly decreased from (1.82 ± 1.08) µg/L to (1.45 ± 0.81) µg/L (P < 0.01) and the level of hs-CRP decreased from (3.40 ± 3.64) mg/L to (1.86 ± 2.02) mg/L (P < 0.01) in the treatment group. (2) Safety index was similar between placebo and treatment groups (P > 0.05) and there was no adverse events including allergic reaction, muscle pain in all subjects during the observation period.

CONCLUSIONThe short-term data obtained from this small hyperlipidemia patient cohort suggest that policosanol is a safe lipid-lowering and anti-inflammatory agent for hyperlipidemia patients.

Anti-Inflammatory Agents ; adverse effects ; therapeutic use ; Anticholesteremic Agents ; adverse effects ; therapeutic use ; Fatty Alcohols ; adverse effects ; therapeutic use ; Female ; Heme Oxygenase-1 ; metabolism ; Humans ; Hyperlipidemias ; blood ; drug therapy ; Lipids ; blood ; Male ; Middle Aged

The clinical efficacy and safety of auricular acupuncture (AA) for treatment of primary insomnia was evaluated. After a comprehensive retrieval in domestic and foreign databases, literatures were strictly screened and Revman 5.2 software was applied to perform a Meta-analysis on eligible randomized controlled trials (RCTs). The evidence quality was assessed with GRADE profiler 3.6 software. As a result, 8 articles were included involving 894 patients. Compared among AA and sham AA, placebo AA, blank control, there was significant difference in Pittsburgh sleep quality index (PSQI) [WMD = -3.48, 95% CI (-3.96, -3.00)], sleep latency LWMD = -10.14, 95% CI (-17.16, -3.12)] and sleep awakening times [WMD = -9.98, 95% CI (-1.10,-0.48)]. Compared between AA and western medication, there was significant difference in PSQI [WMD = -3.62, 95% CI (-4.59, -2.65)]. The evidence quality was moderate in AA vs. sham AA, placebo AA or blank control, while that of the rest was extremely low. No reports of adverse events were described in all studies. In conclusion, for the treatment of primary insomnia, AA could effectively improve sleep quality, but due to the low evidence quality, cautious attitude should be taken on this conclusion, and clinical trials with large sample and high quality were needed in the further.
Acupuncture, Ear ; Humans ; Randomized Controlled Trials as Topic ; Sleep ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy

Adult ; Aged ; Antimalarials ; therapeutic use ; Female ; Humans ; Immune Reconstitution Inflammatory Syndrome ; drug therapy ; JC Virus ; immunology ; Leukoencephalopathy, Progressive Multifocal ; drug therapy ; Male ; Mefloquine ; therapeutic use ; Middle Aged ; Treatment Outcome

Objective To establish an isolated rat pancreas perfusion technique,a method for the precise measurement of insulin secretion in vitro.Methods An isolated rat pancreas perfusion technique was applied in the study of insulin secretion from?-cells in 10 high-fat diet-induced obese Wistar rats.Results For the assessment of the functional integrity of the perfused pancreas,the isolated pancreas of 6 rats met all the criteria: (1)The constancy of perfusion pressure was kept over the whole experiment time[(70?5)mm Hg,1 mm Hg= 0.133 kPa].(2)The duodenal peristaltic activity of isolated pancreas and duodenum block was present after perfusion experiment.(3)Total insulin response to arginine stimulation was significantly increased as compared with glucose stimulation[maximum insulin secretion rate:(987?100)?U/min vs(545?50)?U/min,P