Detection of an anti-angina therapeutic module in the effective population treated by a multi-target drug Danhong injection: a randomized trial.

It's a challenge for detecting the therapeutic targets of a polypharmacological drug from variations in the responsed networks in the differentiated populations with complex diseases, as stable coronary heart disease. Here, in an adaptive, 31-center, randomized, double-blind trial involving 920 patients with moderate symptomatic stable angina treated by 14-day Danhong injection(DHI), a kind of polypharmacological drug with high quality control, or placebo (0.9% saline), with 76-day following-up, we firstly confirmed that DHI could increase the proportion of patients with clinically significant changes on angina-frequency assessed by Seattle Angina Questionnaire (ΔSAQ-AF ≥ 20) (12.78% at Day 30, 95% confidence interval [CI] 5.86-19.71%, P = 0.0003, 13.82% at Day 60, 95% CI 6.82-20.82%, P = 0.0001 and 8.95% at Day 90, 95% CI 2.06-15.85%, P = 0.01). We also found that there were no significant differences in new-onset major vascular events (P = 0.8502) and serious adverse events (P = 0.9105) between DHI and placebo. After performing the RNA sequencing in 62 selected patients, we developed a systemic modular approach to identify differentially expressed modules (DEMs) of DHI with the Zsummary value less than 0 compared with the control group, calculated by weighted gene co-expression network analysis (WGCNA), and sketched out the basic framework on a modular map with 25 functional modules targeted by DHI. Furthermore, the effective therapeutic module (ETM), defined as the highest correlation value with the phenotype alteration (ΔSAQ-AF, the change in SAQ-AF at Day 30 from baseline) calculated by WGCNA, was identified in the population with the best effect (ΔSAQ-AF ≥ 40), which is related to anticoagulation and regulation of cholesterol metabolism. We assessed the modular flexibility of this ETM using the global topological D value based on Euclidean distance, which is correlated with phenotype alteration (r2: 0.8204, P = 0.019) by linear regression. Our study identified the anti-angina therapeutic module in the effective population treated by the multi-target drug. Modular methods facilitate the discovery of network pharmacological mechanisms and the advancement of precision medicine. (ClinicalTrials.gov identifier: NCT01681316).

Discovery of the active compounds of Smilacis Glabrae Rhizoma by utilizing the relationship between the individual differences in blood drug concentration and the pharmacological effect in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: This study addresses the rapid discovery of the active compounds (the original constituents and/or metabolites) of a traditional Chinese drug, Smilacis Glabrae Rhizoma (SGR). AIM OF THE STUDY: The aim of this study was to develop a new method to find out the active compounds of traditional drugs in vivo. MATERIALS AND METHODS: A method was established to discover and identify the potential active compounds in drug-containing plasma from rats that were orally administered SGR extract, utilizing the relationship between the individual differences in blood drug concentrations in the rats and the resulting differences in pharmacological effect, and the method was denoted as the RID-PE method. For this method, we used high-performance liquid chromatography with a diode array detector combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry (LC-MSn) to identify the compounds (the original constituents and metabolites) and to determine the peak areas of the compounds in drug-containing plasma following SGR treatment. The anti-inflammatory effect of SGR was evaluated using a carrageenan-induced inflammatory rat model. According to the percent inhibition of paw edema in each model rat (14 rats total) orally administered SGR extract, the plasma samples from the rats were sorted and divided into 7 groups. Each group consisted of two plasma samples, and their percent inhibition of paw edema were similar to each other. We performed an LC-MSn analysis on 3 plasma groups, which showed large differences in the inhibition rates, with percent inhibitions of 92.7%, 72.4% and 38.4%. The correlation coefficients (r) between the peak area of each compound and the pharmacological effect (inhibition ratio) of SGR in the three groups were analyzed using SPSS software. When the correlation coefficients of the compounds are greater than 0.8 (0.8 < r ≤1), these compounds are strongly and positively correlated with anti-inflammatory activity, making them potential anti-inflammatory active compounds. RESULTS: Fifty-eight potential anti-inflammatory compounds (0.8 < r ≤ 1) from SGR were discovered in model rat plasma using the RID-PE method, 47 of which were considered to be new potentially anti-inflammatory compounds. Among these compounds, four original constituents and 5 isomers of potential anti-inflammatory metabolites were validated to have significant anti-inflammatory effects, and they included astilbin, syringic acid, catechin, coumalic acid, resveratrol-3'-O-glucuronide (RG, isomer of M2 or M3), 3'-O-methyl-(+)-epicatechin-4'-O-glucuronide (CA-1, isomer of M16), 4'-O-methyl-(+)-epicatechin-3'-O-glucuronide (CA-2, isomer of M16), 4'-O-methyl-(+)-epicatechin-7-O-glucuronide (CA-3, isomer of M16) and 3'-O-methyl-(+)-epicatechin-7-O-glucuronide (CA-4, isomer of M16). In addition, four isomers (CA-1-CA-4) were reported to have anti-inflammatory effects for the first time, and CA-3 was a new compound. CONCLUSIONS: The RID-PE method can be used to discover and identify the active constituents and metabolites of SGR systematically and in vivo. Furthermore, these findings enhance our understanding of the metabolism and effective forms of SGR.

[One stage three-column osteotomy for the treatment of scoliosis with split spinal cord malformation].

OBJECTIVE: To analyze the effectiveness and safety of one stage three column osteotomy in treatment of scoliosis with split spinal cord malformation. METHODS: The clinical data of 41 patients with scoliosis and split spinal cord malformation underwent one-stage three-column osteotomy from January 2015 to December 2017 were retrospectively analyzed. There were 17 males and 24 females with average age of (25.14±4.51) years old and the average weight of (65.14±9.11) kg. According to the classification of longitudinal spina bifida, 15 cases of Pang typeⅠwere group A and 26 cases of Pang typeⅡwere group B. The general situations of two groups were recorded ; preoperative and postoperative Cobb angle were observed and the correction rate of Cobb angle of coronal plane was calculated ; the coronal and sagittal torso offset distances were compared between two groups and the trunk balance was evaluated ; the complication of two groups was recorded. RESULTS: All 41 patients were followed up for more than 12 months. The operation time, intraoperative blood loss, and perioperative blood transfusion volume in group A were (610.14±115.02) min, (4 001.12±1 014.33) ml, (3 951.14±1 021.55) ml, respectively, and group B were (520.12±101.14) min, (2 701.57±1 021.45) ml, (2 565.77±880.47) ml, the difference between the two groups was statistically significant (P<0.05). The postoperative hospital stays in the group A and B were (9.45±4.21) days and (9.14±3.01) days, respectively, and there was no significant difference (P>0.05). There was no significant difference in postoperative coronary Cobb angle and correction rate between two groups (P>0.05). Immediately after surgery and 12 months after surgery, there was no significant difference in the trunk displacement distance of coronal view and sagittal view between two groups (P>0.05). Six patients in group A had complications, which was higher than that in group B of 1 case (χ2=4.885, P< 0.05). CONCLUSION: One-stage three-column osteotomy in treatment of scoliosis with split spinal cord malformation has high correction rate and good balance of the trunk. However, for patients with typeⅠsplit spinal cord malformation, they will face longer operation time, more intraoperative bleeding volume, large amount of perioperative blood transfusion and higher risk of complications, and the safety is not as good as that of typeⅡpatients. Therefore, in the actual treatment of scoliosis, especially for those with typeⅠsplit spinal cord malformation, a more reasonable surgical plan should be developed in combination with the actual situations of the patients, so as to improve the safety of the operation.

Detection of 191 Taxifolin Metabolites and Their Distribution in Rats Using HPLC-ESI-IT-TOF-MS(n).

Taxifolin is a ubiquitous bioactive constituent of foods and herbs. To thoroughly explore its metabolism in vivo, an HPLC-ESI-IT-TOF-MS(n) method combined with specific metabolite detection strategy was used to detect and identify the metabolites of taxifolin in rats. Of the 191 metabolites tentatively identified, 154 were new metabolites, 69 were new compounds and 32 were dimers. This is the first report of the in vivo biotransformation of a single compound into more than 100 metabolites. Furthermore, acetylamination and pyroglutamic acid conjugation were identified as new metabolic reactions. Seventeen metabolites were found to have various taxifolin-related bioactivities. The potential targets of taxifolin and 63 metabolites were predicted using PharmMapper, with results showing that more than 60 metabolites have the same five targets. Metabolites with the same fragment pattern may have the same pharmacophore. Thus these metabolites may exert the same pharmacological effects as taxifolin through an additive effect on the same drug targets. This observation indicates that taxifolin is bioactive not only in the parent form, but also through its metabolites. These findings enhance understanding of the metabolism and effective forms of taxifolin and may provide further insight of the beneficial effects of taxifolin and its derivatives.

The profiling and identification of the metabolites of 8-prenylkaempferol and a study on their distribution in rats by high-performance liquid chromatography with diode array detection combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry.

8-Prenylkaempferol is a prenylflavonoid that has various bioactivities and benefits for human health. A high-performance liquid chromatography with a diode array detector combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry (HPLC-DAD-ESI-IT-TOF-MS(n) ) method was established to profile and identify the metabolites of 8-prenylkaempferol in rat in vivo and in vitro, and to study the distribution of these metabolites in rats for the first time. A total of 38 metabolites were detected and tentatively identified, 30 of which were identified as new compounds. The new in vivo metabolic reactions in rats of prenylflavonoids of isomerization, polymerization, sulfation, amino acid conjugation, vitamin C conjugation and other known metabolic reactions were found in the metabolism of 8-prenylkaempferol. The numbers of detected metabolites in feces, urine, plasma, small intestine, stomach, kidneys, liver, heart, lungs, spleen and hepatic S9 fraction were 31, 19, 1, 20, 13, 8, 7, 3, 3, 1 and 11, respectively. This indicated that small intestine and stomach were the major organs in which the 8-prenylkaempferol metabolites were distributed. Furthermore, 16 metabolites were determined to have bioactivities based on the literature and 'PharmMapper' analysis. These findings are useful for better comprehension of the effective forms, target organs and pharmacological actions of 8-prenylkaempferol. Moreover, they provide a reference for the study of the metabolism and distribution of prenylflavonoid aglycone compounds.

[Treatment of Chronic Heart Failure Patients with Qi-Yang Deficiency and Blood Stasis Resistance Syndrome by Xnmallong Injection: a Multi-center Randomized Control Study].

OBJECTIVE: To evaluate the efficacy and safety of Xinmailong Injection (XI) in treatment of chronic heart failure (CHF) patients with qi-yang deficiency and blood stasis resistance syndrome (QY-DBSRS). METHODS: Totally 238 CHF patients with QYDBSRS were assigned to the treatment group (118 cases) and the control group (120 cases) by randomized, double-blind, placebo parallel controlled method. Patients in the treatment group received routine therapy and XI (100 mg/2 mL, by dripping at 5 mg/kg, twice per day for 5 consecutive days), while those in the control group received routine therapy and XI mimetic agent (100 mg/2 mL, by dripping at 5 mg/kg, twice per day for 5 consecutive days). The heart function classification of New York Heart Association (NYHA), 6-min walking distance, left ventricular ejection fraction (LVEF), scores for Chinese medical symptoms were observed before and after treatment, and safety assessed. RESULTS: Totally 235 patients actually entered full analysis set (FAS), including 120 cases in the control group and 115 cases in the treatment group. The total effective rate of heart function, 6-min walking distance and increased post-pre-treatment distance in the experimental group were superior to those of the control group with statistical difference (all P < 0.05). Compared with the control group, increased value of post-pre-treatment LVEF, the total effective rate of Chinese medical syndrome efficacy, scores for Chinese medical symptoms and decreased post-pre-treatment value of Chinese medical syndrome scores were obviously improved (all P < 0.05). There was no significant difference in the incidence of adverse events between the two groups (P > 0.05). CONCLUSIONS: XI could improve the heart function of CHF patients, improve Chinese medical symptoms, elevate exercise tolerance, and improve LVEF. It had no obvious toxic and side effects.

[Simultaneous determination of seven bioactive constituents in Smilacis Glabrae Rhizoma by high-performance liquid chromatography].

This study is to develop an HPLC method for the simultaneous determination of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoisoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in Smilacis Glabrae Rhizoma. Samples of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma were separated on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL · min(-1). The detected wavelength was set at 230 nm and the column temperature was maintained at 35 °C. The contents of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in 84 Smilacis Glabrae Rhizoma samples were in the range of trace-1.381, trace-9.913, trace-3.673, 0.6706-27.08, trace-1.181, trace-4.833 and trace-2.754 mg · g(-1), respectively. The established method was used for analysis of common adulterants. The results demonstrated that the contents of (-)-epicatechin in Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma were 0.01163-0.06007 mg · g(-1) and 0.01583-0.08585 mg · g(-1), respectively, while the other six constituents were not detected. The method is simple and accurate, and can be used for the quality control of Smilacis Glabrae Rhizoma. The constituents of Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma are significantly different from Smilacis Glabrae Rhizoma, and they are not suitable for using as Smilacis Glabrae Rhizoma.