Extending the Grading of Recommendations Assessment, Development and Evaluation (GRADE) in Traditional Chinese Medicine (TCM): The GRADE-TCM.

AIM: To extend and form the "Grading of Recommendations Assessment, Development and Evaluation in Traditional Chinese Medicine" (GRADE-TCM). METHODS: Methodologies were systematically reviewed and analyzed concerning evidence-based TCM guidelines worldwide. A survey questionnaire was developed based on the literature review and open-end expert interviews. Then, we performed expert consensus, discussion meeting, opinion collection, external examination, and the GRADE-TCM was formed eventually. RESULTS: 265 Chinese and English TCM guidelines were included and analyzed. Five experts completed the open-end interviews. Ten methodological entries were summarized, screened and selected. One round of consensus was conducted, including a total of 22 experts and 220 valid questionnaire entries, concerning 1) selection of the GRADE, 2) GRADE-TCM upgrading criteria, 3) GRADE-TCM evaluation standard, 4) principles of consensus and recommendation, and 5) presentation of the GRADE-TCM and recommendation. Finally, consensus was reached on the above 10 entries, and the results were of high importance (with voting percentages ranging from 50 % to 81.82 % for "very important" rating) and strong reliability (with the Cr ranging from 0.93 to 0.99). Expert discussion meeting (with 40 experts), opinion collection (in two online platforms) and external examination (with 14 third-party experts) were conducted, and the GRADE-TCM was established eventually. CONCLUSION: GRADE-TCM provides a new extended evidence-based evaluation standard for TCM guidelines. In GRADE-TCM, international evidence-based norms, characteristics of TCM intervention, and inheritance of TCM culture were combined organically and followed. This is helpful for localization of the GRADE in TCM and internationalization of TCM guidelines.

[Effects of density on growth and gene transcription characteristics of Rehmannia glutinosa].

The present study analyzed the effects of planting density on the development, quality, and gene transcription characte-ristics of Rehmannia glutinosa using 85-5 and J9 as materials with three planting densities of 5 000, 25 000, and 50 000 plants/Mu(1 Mu≈667 m~2). The agronomic characteristics of leaves and tuberous roots, the content of catalpol and acteoside, and the changes of gene expression were determined. The results showed that the leaf size, the diameter of tuberous root, leaf biomass, tuberous root number, and tuberous root biomass per plant at low density were significantly higher than those of medium and high densities. The content of catalpol and acteoside in leaves was higher at high density. The content of catalpol in tuberous roots was higher at low density, and the change trend was similar to that in leaves, while the content of acteoside in tuberous roots was higher at high density. Transcriptome analysis found that about 1/2 of the expansin genes could change regularly in response to density treatment, which was rela-ted to the development of tuberous roots. The change trend of the gene expression of multiple catalytic enzymes involved in the biosynthesis of catalpol and acteoside was consistent with that of their content, which was presumedly involved in the accumulation and regulation of density-responsive medicinal components. Based on the analysis of the development, medicinal components, and gene expression characteristics of R. glutinosa at different densities, this study is expected to provide an important basis for regulating the quality and yield of medicinal materials of R. glutinosa by managing the planting density.

[Effect and mechanism of moxibustion on oxidative stress injury in rats with Parkinson's disease by activating Nrf2/ARE pathway].

OBJECTIVE: To observe the effect of moxibustion on oxidative stress injury of nigrostriatal system in rats with Parkinson's disease (PD) based on nuclear factor erythroid 2-related factor (Nrf2)/antioxidant response element (ARE) pathway, and to explore its mechanism. METHODS: A total of 48 SD rats were randomized into a blank group, a sham-operation group, a model group and a moxibustion group, 12 rats in each group. Unilateral two-point injection with 6-hydroxydopamine (6-OHDA) was adopted in the model group and the moxibustion group to establish the PD model; the operation manipulation in the sham-operation group was the same as the model group and the moxibustion group, and the same volume of 0.9% sodium chloride solutions was given by unilateral two-point injection. Moxibustion was adopted at "Baihui" (GV 20) and "Sishencong" (EX-HN 1) in the moxibustion group for 20 min, once a day, 6 times a week for 6 weeks. No intervention was given in the other 3 groups. Morphology of right mesencephalon substantia nigra was observed by HE staining, the expression of tyrosine hydroxylase (TH) in right mesencephalon substantia nigra was detected by immunohistochemistry method, the expression of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-Px) in corpus striatum was detected by colorimetry method, and the expression of Nrf2 and heme oxygenase-1 (HO-1) proteins was detected by Western blot in the 4 groups. RESULTS: Clear tissue structure and complete dopaminergic neurons of right mesencephalon substantia nigra were observed in the blank group and the sham-operation group; unclear tissue structure, decreased and swelling dopaminergic neurons were observed in the model group; compared with the model group, more neurons were observed and the swelling of cyton was reduced in the moxibustion group. Compared with the sham-operation group, the expression of TH in right mesencephalon substantia nigra was decreased in the model group (P<0.01); compared with the model group, the expression of TH in right mesencephalon substantia nigra was increased in the moxibustion group (P<0.05). Compared with the sham-operation group, the expression of ROS, MDA was increased (P<0.01), the expression of GSH, GSH-Px, Nrf2 and HO-1 was decreased in the model group (P<0.01, P<0.05); compared with the model group, the expression of ROS, MDA was decreased (P<0.05, P<0.01), the expression of GSH, GSH-Px, Nrf2 and HO-1 was increased in the moxibustion group (P<0.05, P<0.01). CONCLUSION: Moxibustion can alleviate oxidative stress injury of nigrostriatal system in rats with Parkinson's disease by activating the Nrf2/ARE pathway, and protect the dopamine neurons.

[Comparative observation of acupoint thread-embedding and acupuncture therapy of Shugan Tiaoshen for insomnia of liver depression and qi stagnation].

OBJECTIVE: To compare the effect of Shugan Tiaoshen (soothing the liver and regulating mind) acupoint thread-embedding therapy and Shugan Tiaoshen acupuncture therapy for insomnia of liver depression and qi stagnation. METHODS: A total of 68 patients with insomnia of liver depression and qi stagnation were randomly divided into an acupoint embedding group (34 cases) and an acupuncture group (34 cases). Two groups were respectively treated with acupoint thread-embedding and acupuncture at Baihui (GV 20), Shenmen (HT 7), Neiguan (PC 6), Taichong (LR 3), Ganshu (BL 18) and Xinshu (BL 15). The treatment in the acupoint embedding group was given once a week, and in the acupuncture group was given once every other day for 6 weeks. The Pittsburgh sleep quality index (PSQI), self-rating anxiety scale (SAS), self-rating depression scale (SDS), TCM syndrome scores before and after treatment and medical expenses were compared between the two groups, and the clinical efficacy was evaluated. RESULTS: After treatment, the PSQI total scores, each sub-item score and SAS and SDS scores of the two groups were lower than those before treatment (P<0.05); the sleep quality, time during fall asleep, sleep time, sleep efficiency, daytime dysfunction scores and PSQI total score in the acupoint embedding group after treatment were lower than those in the acupuncture group (P<0.05). After treatment, except for constipation and yellowish urine, the sub-item scores and total scores of TCM syndromes in the two groups were lower than those before treatment (P<0.05); the sleep disturbance score of the acupoint embedding group was lower than that of the acupuncture group (P<0.05). The medical expenses in the acupoint embedding group was lower than that in the acupuncture group (P<0.05). The total effective rate of the acupoint embedding group was 85.3% (29/34), and the acupuncture group was 82.4% (28/34), there was no significant difference between the two groups (P>0.05). CONCLUSION: Shugan Tiaoshen acupoint thread-embedding therapy and Shugan Tiaoshen acupuncture therapy both have significant curative effect on insomnia of liver depression and qi stagnation, but acupoint thread-embedding therapy is better than acupuncture therapy in improving sleep quality and reducing medical expenses.

[Molecular cloning and expression analysis of iridoid synthase genes from Rehmannia glutinosa].

Iridoid synthase( IS),the key enzyme in the natural biosynthesis of vegetal iridoids,catalyzes the irreversible cyclization of 10-oxogeranial to epi-iridodial. In this study,we screened the Rehmannia glutinosa transcriptome data by BLASTn with Catharanthus roseus CrIS cDNA,and found four c DNA fragments with length of 1 527,1 743,1 425,1 718 bp,named RgIS1,RgIS2,RgIS3 and RgIS4,respectively. Bioinformatics analysis revealed that the four iridoid synthase genes encoding proteins with 389-392 amino acid residues,protein molecular weights were between 44. 30-44. 74 k Da,and theoretical isoelectric points were between 5. 30 and 5. 87. Subcellular localization predictions showed that the four iridoid synthase were distributed in the cytoplasm. Structure analysis revealed that R. glutinosa iridoid synthases contain six conserved short-chain dehydrogenase/reductase( SDR) motifs,and their 3 D models were composed typical dinucleotide-binding " Rossmann" folds covered by helical C-terminal extensions. Using the amino acid sequences of four R. glutinosa iridoid synthases,phylogenetic analysis was performed,the result indicated that RgIS3,CrIS and Olea europaea OeIS were grouped together,the other R. glutinosa iridoid synthases and fifteen proteins in other plants had close relationship. Real-time fluorescent quantitative PCR revealed that RgIS1 and RgIS3 highly expressed in unfold leaves,however,RgIS2 and RgIS4 highly expressed in stems and tuberous roots,respectively. RgIS3 showed higher expression levels in non-radial striations( nRS) of the two cultivars,and RgIS1 and RgIS2 had higher expression levels in nRS of QH,while RgIS4 had less expression levels in nRS of QH1. RgIS1,RgIS2 and RgIS3 were up-regulated by Me JA treatment,although the time and degree of response differed. Our findings are helpful to reveal molecular function of R. glutinosa iridoid synthases and provide a clue for studing the molecular mechanism of iridoid biosynthesis.

Exploration on feasibility of introducing bioassay method into quality evaluation of Chinese herbal medicines by studying on the correlation between antioxidant activity of Prunella vulgaris and its total phenolic acids content for example / 中国中药杂志

This paper aims to investigate the correlation between the antioxidant activity of Prunella vulgaris and its total phenolic acids content by measuring the antioxidant activity of different sources and different organs of P. vulgaris and the total contents of protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid in these samples. Using the 50% methanol extract of P. vulgaris samples as the research object, DPPH method and HPLC method were used respectively to determine the antioxidant activities and the total contents of the above-mentioned five analytes in P. vulgaris samples. 0.5 mL of 50% methanol extract of P. vulgaris reacts with 0.1 mmol•L⁻¹ DPPH ethanol solution for 60 min, then the absorbance of the reaction solution was measured at 517 nm, scavenging rate and IC₅₀ values were calculated by the absorbance and the sample concentration for evaluating the antioxidant activity. HPLC analysis was made on a C₁₈ Epic column, with acetonitrile-0.1% formic acid aqueous solution as mobile phase (gradient elution), and the detection wavelength was set at 280 nm. The correlation between the antioxidant capacity of different habitats and different organs of P. vulgaris and the total contents of five kinds of phenolic acids was analyzed by partial least squares method. The reaction dose-response range of 50% methanol extract of P. vulgaris with 0.1 mmol•L⁻¹ DPPH ethanol solution was 0.300-1.65 g•L⁻¹. When the quantities of potocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid were respectively in 0.007 84-0.980, 0.011 5-1.44, 0.008 64-1.08, 0.080 0-1.00 and 0.079 8-0.998 μg range, their quantities were in good linear relationship with the corresponding peak areas. The average recovery of 5 components were 97.76%, 96.88%, 100.3%, 102.1%, 104.5%, with RSD of 1.8%, 1.6%, 1.7%, 1.6% and 1.7%, respectively. In a certain range of crude drug quantity, the antioxidant activity of each organ of P. vulgaris and total phenolic acids content inside has a good linear correlation. Therefore, in certain quality range of crude drug, DPPH bioassay combined with HPLC content determination can be used for the quality control of P. vulgaris, as is a new method for the quality control of P. vulgaris.

Study on UPLC fingerprint of red ginseng based on good separation and good purity of chromatographic peaks / 中国中药杂志

This study is to establish the UPLC fingerprint of red ginseng. The separation was performed on a Waters Acquity BEH C₁₈ column (2.1 mm × 50 mm,1.7 μm), with the mobile phase consisting of acetonitrile and water for gradient elution. The detection wavelength was set at 203 nm. The UPLC fingerprint of red ginseng was established by using sample chromatography of 22 different purchase areas and 26 common peaks were found. Compared with the reference substances, 11 of the common peaks were identified as ginsenosides Rg₁, ginsenoside Re, ginsenoside Rf, ginsenoside Rh₁, ginsenoside Rg₂, ginsenoside Rb₁, 20(S)-ginsenoside F₁, ginsenoside Rb₂, ginsenoside Rb3, 20(S)-ginsenoside Rg₃ and 20(R)-ginsenoside Rg₃, respectively. It is worth noting that 20(S)-ginsenoside Rg₃ and 20(R)-ginsenoside Rg₃ are the characteristic ingredients of red ginseng, and they could be used not only for distinguishing red ginseng and ginseng, but also for process controlling of the preparation of red ginseng. The similarity was analyzed with' Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica, and the similarity of 18 batches samples is up to 0.9. Compared to the literature methods, the method is simple, time-saving,specific for the separation of ginsenosides from red ginseng. So, this method could be used for the species identification and quality control of ginseng, red ginseng and American ginseng, and it will alsoprovide a theoretical basis of raising quality standards of the above mentioned Chinese herb medicines.

Catgut implantation at acupoints for allergic rhinitis: a systematic review.

OBJECTIVE: To assess the effectiveness and the possible adverse effects of catgut implantation at acupoints for allergic rhinitis (AR). METHODS: This systematic review was carried out in accordance with the Cochrane Handbook version 5.1.0 and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Extensive literature searches were conducted in PubMed, Excerpta Medical Databases, the Cochrane Library, the China National Infrastructure, Wanfang Chinese Digital Periodical and Conference Database, and the Weipu Chinese Science and Technique Journals Database. The Chinese Clinical Trial Registry Center was also searched for ongoing trials up to September 2012. Randomized controlled trials (RCTs) or quasi-RCTs were included. Risk of bias assessment was performed using the Cochrane tool for assessing risk of bias. RESULTS: Five RCTs with 285 participants were found from 49 relevant studies, but there was just one RCT which met the inclusion criteria for this review. The study showed that treatment of catgut implantation at acupoints could lead to a better alleviation of the signs and symptoms of AR than the crude herb moxibustion. No adverse events were reported in this study. CONCLUSIONS: Because of the methodological shortcoming and the risk of bias of the included trial, catgut implantation was proved with only limited evidence for the treatment of AR. Robust RCTs with high quality and larger sample size in this field are hoped to be carried out in the future.

[Effect of ru'ai shuhou recipe on immune response in HER2/neu tranagenic mice undergoing breast cancer carcinogenesis process].

OBJECTIVE: To explore the immune response induced by HER2/neu oncogene in the breast cancer (BC) carcinogenesis process and the immunological mechanism of Ru'ai Shuhou Recipe (RSR) in the prevention and treatment of BC. METHODS: HER2/neu transgenic spontaneous breast tumor model mice were fed with RSR from 5 weeks old, the occurrence of breast tumor in them was observed, and the changes of T cell-mediated immune response and associated cytokines were detected during the carcinogenesis process, i. e., when mice aged between 15 and 25 weeks. RESULTS: RSR showed significant effects in postponing and reducing the carcinogenesis of primary breast tumor, up-regulating the amount of T cell in splenic lymphocyte in tumor-bearing mice, promoting the proliferation of T lymphocyte, and inducing the secretion of cytokines such as interleukin-2, interleukin-12 and interferon-y. CONCLUSIONS: A serial immune response reveals in the carcinogenesis process. The immunologic function of HER-2/neu transgenic mice is significantly different to that of the same strain non-transgenic mice. Effect of RSR in preventing and postponing breast cancer carcinogenesis is possibly realized through enhancing the anti-tumor immune response of transgenic mice themselves.