[Variations of glucose content in Massa Medicata Fermentata during processing based on quantitative proton nuclear magnetic resonance].

A quantitative proton nuclear magnetic resonance(qHNMR) method was established to determine the glucose content in commercially available Massa Medicata Fermentata(MMF) products and explore the variations of glucose content in MMF products during processing. The qHNMR spectrum of MMF in deuterium oxide was obtained with 2,2,3,3-d_4-3-(trimethylsilyl) propionate sodium salt as the internal standard substance. With the doublet peaks of terminal hydrogen of glucose with chemical shift at δ 4.65 and δ 5.24 as quantitative peaks, the content of glucose in MMF samples was determined. The glucose content showed a good linear relationship within the range of 0.10-6.44 mg·mL~(-1). The relative standard deviations(RSDs) of precision, stability, repeatability, and recovery for determination were all less than 2.3%. The glucose content varied in different commercially available MMF samples, which were associated with the different fermentation days, wheat bran-to-flour ratios, and processing methods. The glucose content in MMF first increased and then decreased over the fermentation time. Compared with the MMF products fermented with wheat bran or flour alone, the products fermented with both wheat bran and flour had increased glucose. The glucose content of bran-fried MMF was slightly lower than that of raw MMF, while the glucose content in charred MMF was extremely low. In conclusion, the qHNMR method established in this study is simple, fast, and accurate, serving as a new method for determining the glucose content in MMF. Furthermore, this study clarifies the variations of glucose content in MMF during processing, which can not only indicate the processing degree but also provide a scientific basis for revealing the fermentation mechanism and improving the quality control of MMF.

[Effect of umbilical moxibustion on phlegm damp constitution and intestinal flora].

OBJECTIVE: To observe the effect of umbilical moxibustion on phlegm damp constitution and intestinal flora, and explore the mechanism of umbilical moxibustion in improving phlegm damp constitution. METHODS: A total of 60 subjects with phlegm damp constitution were randomly divided into an observation group and a control group, 30 cases in each group. The control group was given TCM health guidance, such as diet and exercise; on the basis of the control group, umbilical moxibustion was applied in the observation group, 7 moxa cones each time, 2 times a week for 8 weeks. Before and after intervention, the indexes (transformation score of phlegm damp constitution, body mass, body mass index [BMI], waist circumference, hip circumference, heart rate and blood pressure) related to phlegm damp constitution were recorded in the two groups. The intestinal flora was detected with 16S rDNA sequencing technology in the two groups. RESULTS: After intervention, the transformation score of phlegm damp constitution, body mass, BMI, waist circumference and hip circumference were decreased compared before intervention in the observation group (P<0.001, P<0.01, P<0.05), the transformation score of phlegm damp constitution and waist circumference were decreased compared before intervention in the control group (P<0.001, P<0.05); the transformation score of phlegm damp constitution and waist circumference in the observation group were lower than the control group (P<0.001, P<0.05). After intervention, the relative abundance of Eubacterium hallii in the observation group was increased (P<0.01), the relative abundance of Blautia in the control group was decreased (P<0.05). CONCLUSION: Umbilical moxibustion may reshape the intestinal flora by up-regulating the relative abundance of Eubacterium hallii, thus improving phlegm damp constitution.

Acupuncture for Subthreshold Depression: Study Protocol for a Randomized Controlled Trial.

BACKGROUND: Subthreshold depression refers to a state of mental sub-health that has depressive symptoms but does not meet the diagnostic criteria for major depressive disorder. It is a strong risk factor for depression and is related to an increase in suicide and mortality. Studies suggest that acupuncture may be effective in the treatment of subthreshold depression, but no evidence has proven that the efficacy of acupuncture for subthreshold depression is a specific therapeutic effect or a placebo effect. METHODS: This will be a prospective, randomized, controlled, and double-blind study of 64 patients with subthreshold depression. They will be randomly assigned as verum or minimal acupuncture group members. Patients in the verum acupuncture group will receive acupuncture at the acupoints, and those in the minimal acupuncture group will receive minimal acupuncture at non-acupoints. The interventions will be delivered over an 8-week period. The primary outcome measure will be the Hamilton Depression Scale-17 (HAMD-17). The secondary outcome measures will be the 9-item Patient Health Questionnaire (PHQ-9), 7-item Generalized Anxiety Disorder (GAD-7), and SF-12v2 Health Survey. The assessments will occur at baseline, 4 weeks, 8 weeks, and during a follow-up period. DISCUSSION: The protocol uses a randomized controlled trial to examine the effectiveness of acupuncture for subthreshold depression and to further study the mechanisms of its effect.

Jiaji (EX-B2)-Based Electroacupuncture Preconditioning Attenuates Early Ischaemia Reperfusion Injury in the Rat Myocardium.

BACKGROUND: Acupuncture preconditioning was able to reduce the extent of ischaemia reperfusion (I/R) injury. Previous studies have shown that electroacupuncture (EA) pretreatment at T4-T5 Jiaji (EX-B2) acupoints had cardioprotective effects against myocardial I/R injury. However, the molecular mechanism remains inconclusive. METHODS: Wistar rats were pretreated with electroacupuncture for 7 days at the Neiguan (PC6), T4-T5 Jiaji (EX-B2), Yanglingquan (GB34), and Quchi (LI11) acupoints, which belong to different meridians. Then, we investigated the genome-wide gene expression profiles of rats prestimulated at these acupoints after I/R injury. RESULTS: Our study revealed previously unknown cardioprotective roles of T4-T5 Jiaji (EX-B2) acupoints in the I/R progression. The extent of myocardial injury was significantly decreased in the Jiaji group compared with the I/R group. In addition, our data are among the first to link the EA preconditioning at Neiguan (PC6) acupoints and circadian rhythm in the I/R model. Also, for the first time, we explored the meridian and acupoint specificity involved in EA pretreatment at the heart meridian, in which Yanglingquan and Quchi acupoints were selected as the control group for heart-divergent-meridian and nonheart-meridian acupoints. CONCLUSIONS: The present study suggested that EA pretreatment at Jiaji alters genome-wide gene expression and protects the rat myocardium against I/R injury, which are most likely through neurohumoral regulation.

Salen-based bifunctional chemosensor for copper (II) ions: Inhibition of copper-induced amyloid-ß aggregation.

Disruption of copper homeostasis is associated with a number of severe diseases including Alzheimer's disease (AD), Parkinson's disease (PD), Wilson's disease, and Menkes syndrome. Given this association, the detection and capture of Cu2+ in biological fluids and tissues may provide a new direction for the diagnosis and treatment of related disorders. The current analytical approaches, however, are challenging due to the high cost, complexity, and long time required to prepare and analyze samples. Here, we report a novel salen ligand, namely N,N'-(1,2-phenylene)bis(1-(1H-imidazol-4-yl)methanimine) (pimi), which can readily detect and concurrently capture Cu2+ from aqueous as well as biological mediums. Pimi can selectively and specifically detect Cu2+ from biofluid and cellular samples with rapid ccresponse time (<3 s) and an ultra-sensitive detecting limit (2.7 nM). More importantly, pimi showed excellent environmental tolerance and had a very wide pH range for detecting Cu2+ in a variety of biological samples. Attributed to the strong binding affinity and selectivity towards Cu2+, pimi was found to capture Cu2+ ions from Cu-Aß complexes, thus inhibiting copper-induced aggregation of Aß and protecting neuronal cells from the toxicity of aggregated Aß. These results provide a compelling starting point for further fine-tuning of salen-based chemosensor for the diagnosis and treatment of diseases associated with the hyperaccumulation of copper.

[Effect of "Jiaji" (EX-B2)-electroacupuncture preconditioning on expression of myocardial cytochrome P450 enzyme in rats with acute myocardial ischemia-reperfusion injury].

OBJECTIVE: To investigate the effect of "Jiaji"(EX-B2)-electroacupuncture(EA) preconditioning on structural changes of myocardium and expression of cytochrome P450 (CYP450) enzymes in acute myocardial ischemia-reperfusion injury (MIRI) rats. METHODS: Sixty male Wistar rats were randomly divided into sham operation, model, EX-B2, Neiguan (PC6), Yanglingquan(GB34) and Quchi (LI11) groups (n＝10 in each group). The MIRI model was established by occlusion of the descending anterior branch of the left coronary artery for 30 min, followed by reperfusion for 30 min. EA preconditioning (2 Hz/100 Hz, 1 mA) was respectively applied to EX-B2, PC6, GB34, and LI11 for 30 min, once daily for 7 days. Morphological changes of myocardium were observed by H．E. staining. The ultrastructure of myocardial cells was observed by transmission electron microscope (TEM). The protein expression levels of myocardial CYP450 (CYP1A1, CYP2B2, CYP2C11 and CYP4A2) were detected by Western blot. RESULTS: In MIRI rats, myocardial pathological changes as swollen, disorganized arrangement, and partially ruptured myocardial fibers with unclear limit and necrosis and infiltration of lots of inflammatory cells under microscope, and focal myofilament degeneration, dissolution and necrosis with interstitial edema, vague mitochondria with swelling and vacuolation, ruptured and disorganized arrangement of crests under TEM were found in the ischemic area, which was obviously milder in rats of both EX-B2 and PC6 groups. After modeling, the expression of myocardial CYP1A1 and CYP2B2 protein were significantly up-regulated in the model group relevant to the sham operation group (P<0.05). In EA preconditioning groups, the expression levels of CYP1A1 and CYP4A2 proteins in the EX-B2, PC6 and GB34 groups were obviously down-regulated (P<0.01), and those of CYP2B2 and CYP2C11 proteins in both EX-B2 and PC6 groups were markedly up-regulated relevant to the model group (P<0.01). Compared with the EX-B2 group, the expression levels of CYP1A1, CYP4A2 proteins in the LI11 group were significantly up-regulated (P<0.05), and CYP2B2, CYP2C11 proteins were significantly down-regulated (P<0.05). No significant differences were found between the model and sham operation groups in the expression levels of myocardial CYP2C11 and CYP4A2 proteins, between the LI11 and model groups in the expression of CYP1A1, CYP2B2, CYP2C11 and CYP4A2, between the GB34 and model groups in the expression of CYP2B2, CYP2C11 (all P>0.05)．. CONCLUSION: EA preconditioning at EX-B2 and PC6 can suppress MIRI induced up-regulation of myocardial CYP450 ω-hydroxylase CYP1A1 and CYP4A2 expression and further up-regulate the expression of myocardial epoxygenase CYP2B2 and CYP2C11 proteins in acute MIRI rats, thus playing a role in myocardial protection.

[Effect of "Jiaji"(EX-B 2)-electroacupuncture Preconditioning on Expression of Cytochrome P 450 Signaling Pathways in Myocardial Ischemia-reperfusion Rats].

OBJECTIVE: To investigate the effect of "Jiaji"(EX-B 2)-electroacupuncture(EA) preconditioning on the myocardium and expression of cytochrome P 450 signaling pathways of myocardial cells in myocardial ischemia-reperfusion injury(MI/RI) rats. METHODS: Sixty male Wistar rats were randomly divided into sham operation, model, Jiaji, Neiguan(PC 6), Yanglingquan(GB 34) and Quchi(LI 11) groups (n=10 in each group). MI/RI model was established by occlusion of the descen-ding anterior branch of the left coronary artery. EA preconditioning was respectively applied to EX-B 2, PC 6, GB 34 and LI 11 for 30 min, once daily for 7 days. The ECG-ST segment was detected before and after the modeling. The contents of serum creatine kinase-MB(CK-MB) and lactate dehydrogenase(LDH) were detected by ELISA and microplate method separately. The expression levels of myocardial CYP 450 related genes cyp 1 a 1, cyp 2 b 2, cyp 2 c 11, cyp 4 a 2 were detected by Real-time PCR. RESULTS: Thirty minutes after ligation the changes of ECG-ST segment displacement of the Jiaji group and Neiguan group were signifi-cantly lower than that of the model group(P<0.01), and the change was significantly lower in the Neiguan group than in the model group(P<0.01) 30 minutes after reperfusion. Compared with the sham operation group, the activities of serum CK-MB and LDH were significantly increased in the model group(P<0.01). Compared with the model group, the activities of serum CK-MB and LDH were significantly decreased in the Jiaji, Neiguan and Yanglingquan groups (P<0.01). Compared with the Jiaji group, the activity of serum LDH was significantly increased in the Quchi group, while the activities of serum CK-MB were significantly increased in the Quchi and Yanglingquan groups (P<0.05). Compared with the sham operation group, the expression levels of myocardial cyp 1 a 1 and cyp 2 b 2 mRNAs were significantly higher in the model group(P<0.01), while the expression of cyp 2 c 11 mRNA was lower(P<0.01). Compared with the model group, the expression levels of cyp 1 a 1 and cyp 4 a 2 mRNAs were lower in the Jiaji, Neiguan and Yanglingquan groups (P<0.01), while the expressions of cyp 2 b 2 and cyp 2 c 11 mRNAs were higher(P<0.01). Compared with the Jiaji group, the expression of cyp 1 a 1 mRNA was lower in the Neiguan group, while the expression of cyp 2 b 2 mRNA was higher(P<0.05). CONCLUSIONS: EA preconditioning at Jiaji(EX-B 2) can reduce serum myocardial enzymes of myocardial ischemia-reperfusion rats, lower the ST segment of electrocardiogram, up-regulate myocardial CYP 450 epoxygenase cyp 2 b 2 and cyp 2 c 11 mRNAs expression, down-regulate ω-hydroxylase cyp 1 a 1 and cyp 4 a 2 mRNAs expression, thus play a role in myocardial protection.

[Purine and carboline alkaloids from ascidian Symplegma oceania].

Five purine and carboline alkaloids were isolated from the methanol extract of the ascidian Symplegma oceania. Classic chromatographies including preparative HPLC were used for isolation and purification of the compounds. The structures were established as 6-methoxy-7-methyl-8-oxoguanine (1), 2-methylimino-3-methyl-6-methylamino- 9H-purine (2), 1,2,3,4-tetrahydro-betacarboline (3), 1,2,3,4-tetrahydro-1-methyl-beta-carboline (4) and 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (5) by comparison the spectroscopic data (MS, 1H, 13C-NMR) with those reported in the literatures. Compounds 2-5 were reported from the the genus Symplegma for the first time. The purine and carboline were the major alkaloid types of S. oceania.