Multiomic Analysis Reveals Core Regulatory Mechanisms underlying Steroidal Glycoalkaloid Metabolism in Potato Tubers.

Steroidal glycoalkaloids (SGAs) present in germinated potato tubers are toxic; however, the mechanisms underlying SGA metabolism are poorly understood. Therefore, integrated transcriptome, metabolome, and hormone analyses were performed in this study to identify and characterize the key regulatory genes, metabolites, and phytohormones related to glycoalkaloid regulation. Based on transcriptome sequencing of bud eyes of germinated and dormant potato tubers, a total of 6260 differentially expressed genes were identified, which were mainly responsible for phytohormone signal transduction, carbohydrate metabolism, and secondary metabolite biosynthesis. Two TCP14 genes were identified as the core transcription factors that potentially regulate SGA synthesis. Metabolite analysis indicated that 149 significantly different metabolites were detected, and they were enriched in metabolic and biosynthetic pathways of secondary metabolites. In these pathways, the α-solanine content was increased and the expression of genes related to glycoalkaloid biosynthesis was upregulated. Levels of gibberellin and jasmonic acid were increased, whereas that of abscisic acid was decreased. This study lays a foundation for investigating the biosynthesis and regulation of SGAs and provides the reference for the production and consumption of potato tubers.

Analysis of pathogenic bacteria and influencing factors of death in patients with severe neurological pulmonary infection / 中国医师杂志

Objective:To investigate the distribution of respiratory pathogens and risk factors of death in patients with pulmonary infection in neurosurgical intensive care unit (NICU).Methods:A total of 87 patients with pulmonary infection in the NICU of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine from January 2018 to December 2019 were collected, and the pathogens of their respiratory tract were analyzed to understand the types and distribution of bacteria in the lung infection. Univariate statistical analysis was used to analyze the relationship between the patient′s clinical outcome with age, diabetes, hypertension, renal insufficiency, hypoproteinemia, anemia, chronic respiratory disease, surgery, tracheotomy, and bacterial multi-resistance. Binary logistic regression analysis was used to analyze the influencing factors of death in NICU patients with pulmonary infection.Results:A total of 112 pathogenic bacteria were isolated in this research group, including 83 Gram-negative bacteria (74.11%), 22 Gram-positive bacteria (19.64%), and 7 Fungi (5.25%). Imipenem was highly sensitive to Gram-negative bacteria, vancomycin was highly sensitive to Gram-positive bacteria, and other drugs were highly resistant. 41 patients died (47.13%). Age≥60 ( OR=3.501, 95% CI: 1.152-10.638), renal insufficiency ( OR=3.872, 95% CI: 1.336-11.224), tracheotomy ( OR=0.317, 95% CI: 0.114-0.882), bacteria multi-drug resistance ( OR=3.480, 95% CI: 1.162-10.422) were independent risk factors for death in NICU patients with pulmonary infection. Conclusions:Patients with severe neurological diseases are in critical condition, and there are many patients with pulmonary infection, with poor prognosis and high mortality. Gram-negative bacteria are the most common respiratory pathogens. Carbapenems account for the highest proportion of antibiotics in clinic. Advanced age, renal insufficiency and bacterial multidrug resistance increase the mortality of patients, while early tracheotomy can reduce the mortality of patients.

Characteristic chromatogram and index components content of substance benchmark of Xuanfu Daizhe Decoction / 中国中药杂志

The methods for determining the characteristic chromatogram and index components content of Xuanfu Daizhe Decoction were established to provide a scientific basis for the quality evaluation of substance benchmarks and preparations. Eighteen batches of Xuanfu Daizhe Decoction were prepared with the decoction pieces of different batches and of the same batch were prepared respectively, and the HPLC characteristic chromatograms of these samples were established. The similarities of the chromatographic fingerprints were analyzed. With liquiritin, glycyrrhizic acid, 6-gingerol, ginsenoside Rg_1, and ginsenoside Re as index components, the high performance liquid chromatography was established for content determination with no more than 70%-130% of the mass average as the limit. The results showed that there were 19 characteristic peaks corresponding to the characteristic chromatograms of 18 batches of Xuanfu Daizhe Decoction, including 8 peaks representing liquiritin, 1,5-O-dicaffeoylqunic acid, ginsenoside Rg_1, ginsenoside Re, 1-O-acetyl britannilactone, ginsenoside Rb_1, glycyrrhizic acid, and 6-gingerol, and the fingerprint similarity was greater than 0.97. The contents of liquiritin, glycyrrhizic acid, 6-gingerol, and ginsenosides Rg_1 + Re in the prepared Xuanfu Daizhe Decoction samples were 0.53%-0.86%, 0.61%-1.2%, 0.023%-0.068%, and 0.33%-0.66%, respectively. Except for several batches, most batches of Xuanfu Daizhe Decoction showed stable contents of index components, with no discrete values. The characteristic chromatograms and index components content characterized the information of Inulae Flos, Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, and Zingiberis Rhizoma Recens in Xuanfu Daizhe Decoction. This study provides a scientific basis for the further research on the key chemical properties of substance benchmark and preparations of Xuanfu Daizhe Decoction.

Effects and mechanisms of Supplemented Gegen Qinlian Decoction Formula against podocyte pyroptosis and insulin resistance in model rats with diabetic kidney disease / 中国中药杂志

This study explored the in vivo effects and mechanisms of the modern classical prescription Supplemented Gegen Qinlian Decoction Formula(SGDF) against diabetic kidney disease(DKD). Sixty rats were randomly divided into the normal group, model group, SGDF group, and rosiglitazone(ROS) group. The modified DKD rat model was established by employing the following three methods: exposure to high-fat diet, unilateral nephrectomy, and intraperitoneal injection of streptozotocin(STZ). After modeling, rats in the four groups were treated with double distilled water, SGDF suspension, and ROS suspension, respectively, by gavage every day. At the end of the 6 th week of drug administration, all the rats were sacrificed for collecting urine, blood, and kidney tissue, followed by the examination of rat general conditions, urine and blood biochemical indicators, glomerulosclerosis-related indicators, podocyte pyroptosis markers, insulin resistance(IR)-related indicators, and key molecules in the insulin receptor substrate(IRS) 1/phosphatidylinositol-3-kinase(PI3 K)/serine threonine kinase(Akt) signaling pathway. The results showed that SGDF and ROS improved the general conditions, some renal function indicators and glomerulosclerosis of DKD model rats without affecting the blood glucose(BG). Besides, they ameliorated the expression characteristics and levels of podocyte pyroptosis markers, alleviated IR, and up-regulated the protein expression levels of the key molecules in IRS1/PI3 K/Akt pathway to varying degrees. In conclusion, similar to ROS, SGDF relieves DKD by targeting multiple targets in vivo. Specifically, it exerts the therapeutic effects by alleviating podocyte pyroptosis and IR. This study has preliminarily provided the pharmacological evidence for the research and development of new drugs for the treatment of DKD based on SGDF.

Quality evaluation of Aralia taibaiensis based on spectrum-activity relationship / 中国中药杂志

A spectrum-activity relationship is established with high performance liquid chromatography(HPLC) fingerprints and the in vitro antioxidant activity to improve the quality evaluation system of Aralia taibaiensis. The HPLC profiles of 12 batches of samples were collected, and the similarity evaluation, heat map analysis and principal component analysis were conducted for the chemometric study of the fingerprint data. Combined with grey correlation analysis, the contributions of the common peaks in the fingerprints to the antioxidant activity were clarified, and the important peaks reflecting the efficacy were identified. The results showed that 17 common peaks were found in 12 batches of A. taibaiensis samples, and 6 of them were identified as saponins. Similarity evaluation, heat map analysis and principal component analysis roughly classified the A. taibaiensis herbs into two categories, i.e.,(1) S1-S10, S12 and(2) S11. Twelve batches of samples showed different antioxidant activities in a dose-dependent manner. In particular, S9 had the strongest antioxidant activity, while S11 was the weakest in antioxidant capacity, which was basically consistent with the overall score results. The results of grey correlation analysis demonstrated that the 17 common peaks scavenged DPPH radicals in the following order: X_3>X_(17)>X_4>X_8>X_7>X_(13)>X_2>X_6>X_(11)>X_(10)>X_(16)>X_(12)>X_9>X_5>X_(14)>X_1>X_(15), and scavenged ABTS radicals in the order of X_4>X_3>X_7>X_8>X_2>X_(17)>X_(13)>X_6>X_(16)>X_(11)>X_5>X_(12)>X_(10)>X_9>X_(14)>X_1>X_(15). Among them, X_3, X_4, X_7(araloside C), X_8 and X_(17) were the important peaks reflecting the efficacy of A. taibaiensis, which were basically consistent with those contained in the principal component 1. In this study, the correlation between the HPLC fingerprints of 12 batches of A. taibaiensis and its antioxidant activity provides a reference for the Q-marker screening and quality control of A. taibaiensis.

Effect of electroacupuncture on serum melatonin and dopamine in aged insomnia / 中国针灸

OBJECTIVE@#To observe the clinical effect of electroacupuncture (EA) on aged insomnia, and explore its possible mechanism.@*METHODS@#A total of 60 patients with aged insomnia were randomly divided into an EA group (30 cases) and a sham EA group (30 cases, 1 case dropped off). The patients in the EA group were treated with acupuncture at Baihui (GV 20), Yintang (GV 29), Shenmen (HT 7), Sanyinjiao (SP 6), Xinshu (BL 15) and Shenshu (BL 23), and EA was used at Baihui (GV 20) and Yintang (GV 29), with intermittent wave, 2 Hz in frequency. In the sham EA group, the acupoints and the EA connection acupoints were the same as those in the EA group, 2-3 mm in depth, but no current was connected. The intervention was given 30 min each time, once every other day, 3 times a week for 4 weeks in the both groups. Before and after treatment, the Pittsburgh sleep quality index (PSQI) and Montreal cognitive assessment (MoCA) scale were used to assess sleep quality and cognitive function, and serum melatonin (MT) and dopamine (DA) levels were detected.@*RESULTS@#After treatment, the total score and sub-item scores of PSQI in the EA group were lower than those before treatment (@*CONCLUSION@#Electroacupuncture can improve sleep quality and cognitive function in aged insomnia patients, and its mechanism may be related to regulating serum MT and DA levels.

Inhibitory effect of Qing-Fei-Pai-Du decoction on coronavirus in vitro / 药学学报

Qing-Fei-Pai-Du decoction (QFPDD) is a combination of traditional Chinese medicine and plays an important role in the treatment of coronavirus disease 2019 (COVID-19). This study investigated the inhibitory effect of QFPDD on coronavirus replication and antiviral mechanism. The cytotoxicity of QFPDD was determined by PrestoBlue cell viability assay. Quantitive reverse transcription PCR (qRT-PCR) and immunofluorescence assay (IF) were used to detect the inhibitory effects of QFPDD on coronavirus at RNA and protein levels. qRT-PCR was used to detect the adsorption and penetration of coronavirus after QFPDD treatment. The effects of QFPDD on interferon (IFN) and interferon-stimulated genes (ISGs) were also detected by qRT-PCR. The results showed that QFPDD inhibited coronavirus at RNA and protein levels in a dose-dependent manner at non-toxic concentration, and QFPDD targeted in the early stages of coronavirus infection cycle. Preliminary mechanism studies have shown that QFPDD can directly block the virus entry into the cell by inhibiting virus adsorption, and QFPDD can also play an antiviral role by up-regulating the expression of IFN and ISGs. These results indicate QFPDD as a drug potential to treat coronavirus infection.

Effect of compound houttuynia mixture on influenza virus in vitro / 药学学报

Compound houttuynia mixture belongs to OTC class A medicine, which is made from Houttuynia cordata, Scutellaria baicalensis, Radix Isatidis, Forsythia, and Lonicera. As a kind of compound preparation of traditional Chinese medicine, houttuynia cordata mixture has extensive pharmacological effects, for example, clearing away heat and detoxifying, thus it is used for the sore throat, acute pharyngitis, and tonsillitis with wind-heat syndrome. In this study, the antiviral activity against influenza viruses and the primary mechanism of compound houttuynia mixture was evaluated. The antiviral effect of compound houttuynia mixture was determined by cytopathic effects (CPE), Western blot, quantitive reverse transcription PCR (qRT-PCR), and virus titer assays. The effect of houttuynia mixture on the replication cycle of influenza virus was evaluated by time-of-addition assay. In conclusion, the results showed that the compound houttuynia mixture had a broad-spectrum effect against influenza virus, including the international common influenza virus strains, the drug-resistant strains and the highly pathogenic avian influenza viruses H5N1 and H7N9. It mainly impairs the early stage of the viral replication.

Qualitative and quantitative analysis of Arnebiae Radix and Dictamni Cortex and efficacy study of herbal extracts on allergic contact dermatitis using 3D human reconstructed epidermis / 中草药·英文版

Objective: To evaluate the quality of Arnebiae Radix (AR) and Dictamni Cortex (DC) and study the efficacy of herbal extracts of these two herbs on the treatment of allergic contact dermatitis (ACD). Methods: Qualitative and quantitative analysis of effective components was performed using High Performance Thin Layer Chromatography (HPTLC), High Performance Liquid Chromatography (HPLC), and HPLC-Quadrupole Time of Flight-Mass Spectrometry (HPLC-QTOF-MS). In vitro allergic ACD 3D model was established by incubating 3D reconstructed human epidermis (RHE) with skin sensitizer, potassium dichromate. A total of 65 gene expression that were associated with ACD, which included 24 antioxidant responsive element (ARE) and 41 SENS-IS genes were quantified by qRT-PCR. More than or equal to 10 ARE genes and 18 SENN-IS genes were induced by 1.3-fold, demonstrating the successful establishment of in vitro ACD model. Oil extracts of AR and DC were applied on the in vitro ACD model to study the efficacy. Results: Batch 3 of AR and batch 2 of DC showed presence of all active ingredients with the highest concentrations. Active ingredients of the herbs were extracted using a special oil and formulated into herbal oil extracts. The herbal oil extracts were able to down regulate the induced genes in the in-vitro ACD skin model, bringing the tissue back to homeostatic status. Conclusion: The oil extracts showed the potent efficacy of using AR and DC in ACD treatment. The combination study will be done to optimize the formulation ratio which will be developed into a topical cream.

Predictive analysis of Q-marker of Glycyrrhizae Radix et Rhizoma based on network pharmacology combined with qualitative and quantitative research / 中草药

Objective: To make a preliminary prediction of the Q-marker of Glycyrrhizae Radix et Rhizoma from the perspective of the effectiveness and measurability of chemical components based on the concept of Q-marker of Chinese materia medica. Methods: Based on literature integration and data analysis, the source range of Glycyrrhizae Radix et Rhizoma Q-marker was screened, and the effectiveness of the ingredients was analyzed through network pharmacology. Qualitative and quantitative analysis of 15 batches of Glycyrrhizae Radix et Rhizoma from four places of origin was performed by HPLC. The pattern recognition method was used to screen out the main marker components that caused the differences between groups, which were combined with network pharmacological results to further determine the Q-marker of Glycyrrhizae Radix et Rhizoma. Results: Literature studies had determined that flavonoids and triterpenoids were the main source of Glycyrrhizae Radix et Rhizoma Q-marker; Network pharmacology results showed that liquiritin, glycyrrhizic acid and other components had high connectivity in the "component-target-pathway" network and were the main active components; The fingerprints of 15 batches of Glycyrrhizae Radix et Rhizoma samples were established, and five components, including liquiritin and liquiritin apioside, were identified as the main marker components by PLS-DA analysis; The content determination results of liquiritin, liquiritin apioside, glycyrrhizic acid and glycyrrhetinic acid showed that there were significant differences in the content of ingredients among different production areas. The qualitative and quantitative research on pharmacology combined with network pharmacology revealed that liquiritin, liquiritin apioside, glycyrrhizic acid and glycyrrhetinic acid can be used as Glycyrrhizae Radix et Rhizoma Q-marker. Conclusion: Taking flavonoids and triterpenoids as the source of Q-marker for Glycyrrhizae Radix et Rhizoma, the qualitative and quantitative (measurability) study of Glycyrrhizae Radix et Rhizoma herbs from multiple producing areas combined with network pharmacology (effectiveness) revealed liquiritin, liquiritin apioside, glycyrrhizic acid and glycyrrhetinic acid as the potential Q-marker of Glycyrrhizae Radix et Rhizoma are scientific and reasonable, which provide reference for quality control of Glycyrrhizae Radix et Rhizoma.

Insomnia treated with three acupuncture combinations guided by theory of / 中国针灸

OBJECTIVE@#To compare the clinical therapeutic effect and safety on insomnia among three acupuncture combinations, named @*METHODS@#A total of 90 patients with insomnia were randomized into a Tongdu Tiaowei group (30 cases, 1 case dropped off ), a Tongdu Yangxin group (30 cases, 2 cases dropped off ) and a Tongdu Tiaozang group (30 cases, 1 case dropped off ). Besides electroacupuncture (EA) applied to Baihui (GV 20) and Yintang (GV 29) in all of the three groups, acupuncture was added at Shenmai (BL 62) and Zhaohai (KI 6), Shenmen (HT 7) and Sanyinjiao (SP 6) as well as back-@*RESULTS@#After treatment, the score of each of 6 factors, named sleep quality, sleep latency, sleep time, sleep efficiency, sleep disturbance and daytime dysfunction, as well as the total score of PSQI scale were all reduced as compared with the scores before treatment in the patients of each group (@*CONCLUSION@#All of the three acupuncture combinations, named

Meta-analysis of Zhibitai Capsules combined with statin in reducing blood lipid levels in patients with coronary heart disease / 中国中药杂志

To systematically review the efficacy and safety of Zhibitai Capsules combined with chemical drugs versus chemical drugs alone in regulating blood lipid of patients of coronary heart disease, so as to provide evidence-based reference for clinical treatment. In this study, PubMed, EMbase, Cochrane Library, China Knowledge Network Database(CNKI), Technology Journal Database(VIP) and WanFang Database(WanFang) were retrieved to find the randomized controlled trials(RCT) about therapeutic efficacy of Zhibitai Capsules combined with statins(experimental group)versus statins alone(control group)in the treatment of regulating blood lipid of patients with coronary heart disease. The retrieval time was restricted to be from the inception to October 2019. The data were extracted from the randomized controlled trials. Meta-analysis was conducted by RevMan 5.3 statistical software after quality evaluation by Cochrane 5.1.0 quality evaluation tool(blood lipid level, inflammation indicators, traditional Chinese medicine syndrome score and adverse reactions). A total of 11 RCT were included, involving 1 538 patients. The results of Meta-analysis showed that in terms of decrease of total cholesterol(MD=-0.15,95%CI[-0.25,-0.05],P=0.004), decrease of triglycerides improvement(MD=-0.16,95%CI[-0.23,-0.10],P<0.000 01), decrease of low-density lipoprotein(MD=-0.08,95%CI[-0.15,-0.01],P=0.03), and increase of high-density lipoprotein(MD=0.06,95%CI[0.03,0.10],P=0.000 2), experimental group was better than control group. At the same time, the incidence of adverse reactions were low in the experimental group(OR=0.40,95%CI[0.18,0.85],P=0.02). As a result, in treatment of coronary heart disease, the therapeutic efficacy of Zhibitai Capsules combined with statins is better than statins alone in lowering total cholesterol level, triglyceride level, low-density lipoprotein level, and increasing high-density lipoprotein level. Patients in the experimental group had a low incidence of adverse events, but the heterogeneity was slightly higher, and the result had a poor stability. However, due to the small sample size of studies included, some experimental designs were not perfect, which reduces the recommendation level and evidence intensity of this system evaluation. Therefore, high-quality multi-center, large-sample, randomized, double-blind randomized controlled trials are needed for providing more reliable basis.

The application of emotion nursing of traditional Chinese medicine combined with cognitive behavior intervention in reducing the kinesiophobia of patients with lumbar disc herniation during rehabilitation / 中国实用护理杂志

Objective:To discuss the effect of lumbar disc herniation of motility kinesiophobia during convalescence which intervened by Chinese medicine nursing.Methods:A total of 172 patients with lumbar disc herniation treated at TCM Rehabitation Department at Three Gorges Hospital Affiliated to Chongqing University from May 1, 2017 to January 30, 2019 were selected and divided into control group and intervention group by random digits table method with 86 cases each. The control group was given routine psychological nursing, and the intervention group was given emotional nursing of traditional Chinese medicine combined with cognitive behavior intervention. The scores of anxiety, depression and kinesiophobia, the quality of life, the completion and accuracy of rehabilitation training before and after intervention were compared between the two groups.Results:There was no significant difference in the scores of anxiety, depression and kinesiophobia, the quality of life, the completion and accuracy of rehabilitation training before intervention between the two groups( P>0.05). The scores of anxiety, depression at 1,2,3 weeks after intervention was (54.61±7.23), (49.98±6.55), (45.03±6.21) and (55.16±7.53), (50.96±6.45), (46.31±5.72) in the intervention group, (59.69±7.52), (56.92±6.79), (53.62±7.28) and (60.65±7.96), (56.87±6.80), (52.80±5.92) in the control group, the differences were significant( Fgroup values were 10.157, 9.138, P<0.05). The scores of kinesiophobia, the quality of life at 1,2,3 weeks after intervention was (39.66±4.02), (30.58±4.02), (24.38±3.87) and (43.83±2.74), (56.72±3.81), (75.27±5.83)in the intervention group, (43.71±6.87), (37.83±6.81), (33.95±2.86) and (42.50±3.92), (46.05±3.95), (51.62±4.64) in the control group, the differences were significant( Fgroup values were 30.718, 30.938, P<0.01). The completion and accuracy of rehabilitation training at 1, 2, 3 weeks after intervention was (69.08±1.83), (77.05±2.04), (87.34±3.16) and (60.32±2.19), (70.42±3.22), (83.93±2.74) in the intervention group, (54.23±4.22), (62.72±2.65), (73.51±5.41) and (48.24±5.07), (59.77±4.13), (75.64±3.33) in the control group, the differences were significant( Fgroup values were 28.471, 25.139, P<0.01). Conclusions:Emotion nursing of traditional Chinese medicine combined with cognitive behavioral intervention is helpful to relieve the anxiety and depression of patients with lumbar disc herniation, effectively reduce the kinesiophobia of patients during rehabilitation, improve the quality of life of patients, improve the completion and accuracy of rehabilitation training of patients, and contribute to the improvement of rehabilitation effect and the realization of good treatment outcome.

Thermo- and pH-responsive, Lipid-coated, Mesoporous Silica Nanoparticle-based Dual Drug Delivery System To Improve the Antitumor Effect of Hydrophobic Drugs.

Evodiamine (EVO) and Berberine (BBR), from Euodiae Fructus and Coptidis rhizoma, have been used as an herbal medicine pair in traditional Chinese medicine to exert synergistic antitumor effects against various types of tumor cells. However, their clinical use is limited by their poor solubility and adverse toxic side effects. Mesoporous silica nanoparticles (MSNs) possess excellent properties such as a readily functionalized surface, prominent biocompatibility, and huge specific surface area for loading with hydrophobic and hydrophilic drug. On this basis, a novel temperature- and pH-responsive dual drug delivery platform has been developed, in which lipid-coated MSN@p(NIPAM- co-MA) codelivers EVO and BBR. The results indicate that the nanocarrier improves the efficacy and biocompatibility of the drug pair and maintain desirable drug profiles at the low pH and higher temperature of the tumor microenvironment. The dual drug-loaded MSNs showed excellent synergistic therapy effects in vitro (cytotoxicity, cell migration and invasion, angiogenesis) and in vivo (growth of tumor grafts in mice). Meanwhile, the dual drug-loaded nanoparticles showed lower systemic toxicity than either drug alone, the free drug combination, or Taxol. These results suggest that the temperature- and pH-sensitive lipid-coated MSNs are a promising novel carrier for both hydrophobic and hydrophilic drugs.

Identification and expression analysis of four CIPKs genes from a rare and medicinal plant, Dendrobium officinale / 药学学报

The calcineurin B-like protein (CBL)-interacting protein kinase (CIPK) plays a vital role in the growth, development, and stresses adaptation in plants by interaction with the calcium signaling. In this study, four full length cDNAs of CIPKs genes, namely DoCIPK1, DoCIPK2, DoCIPK3 and DoCIPK4 (GenBank accession No. KT957557, KT957558, KT957559 and KT957560, respectively) were cloned from the rear and medicinal plant, Dendrobium officinale, by rapid amplification of cDNA ends (RACE) for the first time. The corresponding encoded proteins, consisting of 473, 449, 451 and 440 amino acids (aa), respectively, with a molecular weight of 53.50, 50.93, 51.50 and 50.16 kDa, and an isoelectric point (pI) of 7.99, 9.25, 8.81 and 9.11, respectively, shared 70%-90%, 69%-80%, 78%-93%, and 66%-82% identities CIPKs with various plants. Each deduced protein contained a conserved protein kinase domain (respectively at 21 -275, 14-268, 16-271 and 12-266 aa position), a CIPKs family characteristic NAF/FISL domain (respectively at 335-391, 313-370, 310-369 and 305-362 aa position) and some functional motifs. The four DoCIPK proteins, without signal peptide or transmembrane region, were located in the plasma membrane and endoplasmic reticulum at the subcellular level. The three dimensional structure of the proteins were similar to that of Arabidopsis AtCIPK24. DoCIPK1 and DoCIPK3 were respectively clustered in the group E and A of the Arabidopsis and rice CIPK evolutionary tree, while DoCIPK2 and DoCIPK4 belonged to group C. The relative expression of DoCIPK1 showed no significant difference in the leaves and stems, and its transcripts in the roots was 0.35 fold over that in the leaves. The abundance of DoCIPK3 transcripts in the stems and the roots were 3.36 fold and 3.47 fold higher, respectively, than those in the leaves. DoCIPK2 exhibited similar expression pattern to DoCIPK4. Their relative expression in the leaves and the stems had no apparent difference, and the transcript levels were higher in the roots than that in the leaves, with 2.08 fold and 7.86 fold, respectively. Cloning, bioinformatics analyses, and expression patterns of the four DoCIPK genes provide a basis for functional elucidation of these genes further during the physiological responses in D. officinale.

Clinical effects of high frequency repeated transcranial magnetic stimulation therapy on dyskinesia in patients with incomplete spinal cord injury:a Meta-analysis / 中国骨伤

<p><b>OBJECTIVE</b>To systematically evaluate the clinical effect of high frequency repeated transcranial magnetic stimulation(HF rTMS)therapy on dyskinesia in patients with incomplete spinal cord injury.</p><p><b>METHODS</b>Randomized controlled trials(RCTs) about HF rTMS therapy on patients with motor incomplete spinal cord injury were searched electronically in PubMed, Google scholar, Cochrane library, Clinical trial, Medline, Web of science, CNKI, VIP, and Wanfang database before October 2016. Two reviewers independently screened the literatures according to the inclusion and exclusion criteria, as well as extracted the data and assessed the methodological quality. The observed outcomes included ASIA motor score, ASIA lower extremities motor score(LEMS), Modified Ashworth score (MAS), Ten-meter walking test (10MWT) and Walking index for SCI II(WISCI II), and the outcomes were analyzed using RevMan5.2 software provided by the Cochrane information management system.</p><p><b>RESULTS</b>Five RCTs involved 103 patients were included, and 61 patients(experimental group) accepted real rTMS and physical rehabilitation care for SCI, 51 patients(control group) accepted only physical rehabilitation care. There were significant differences in ASIA motor score, LEMS and 10MWT between two groups after HF rTMS therapy (statistics were=2.96,=0.003;=3.04,=0.002;=2.16,=0.03; respectively). When stimulating the leg motor cortex, there was significant difference in MAS between two groups(=2.79,=0.005), and when stimulating the vertex, there was no significant difference(=0.09,=0.93). There was no significant difference in WISCI IIscore after HF rTMS therapy between two groups(=0.90,=0.37).</p><p><b>CONCLUSIONS</b>HF rTMS can raise motor score in patients with incomplete spinal cord injury, improve the spasticity of the lower extremities, and increase the motor ability.</p>

Water-soluble extract of clove inhibits in vitro and in vivo growth of colon cancer by inducing autophagy / 中国药理学与毒理学杂志

OBJECTIVE Cloves(Syzygium aromaticum L.)have been used as both a spice and a traditional Chinese medicinal herb for thousands of years. However, relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti-tumor effects and mechanisms of water extract of cloves(WEC)against colorectal cancer. METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3a/b)were determined by western blot. RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50values of 150 μg·mL-1.The colony-formation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice. Flow cytometry result showed that WEC significantly induced autophagy, and the averaged relative values of fluorescence intensity were 206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I. CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo, which might be related with autophagy induction, and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.

Chemical constituents and anti-inflammatory activities of Maqian (Zanthoxylum myriacanthum var. pubescens) bark extracts.

In this study, 44 compounds in the petroleum ether extract of Maqian (Zanthoxylum myriacanthum var. pubescens) bark, a traditional Dai herbal medicine, were identified by GC-MS. Major components included 3(2H)-benzofuranone, asarinin and (dimethoxymethyl)-3-methoxy-benzene. A total of 18 compounds were isolated from the ethyl acetate extracts of Maqian bark by column chromatography and identified by chemical and spectral analyses. Rhoifoline B, zanthoxyline dimethoxy derivative, N-nortidine, nitidine, decarine are the major alkaloids. Both the petroleum ether and ethyl acetate extracts showed significant inhibition on NO production, which imply anti-inflammatory activity, in lipopolysaccharide-induced RAW 264.7 cells without cell toxicity. Decarine is the major anti-inflammatory constituent with NO IC50 values of 48.43 µM on RAW264.7 cells. The petroleum ether extract, the ethyl acetate extract and decarine showed anti-inflammatory activities through inhibiting TNF-α and IL-1ß production in lipopolysaccharide-stimulated THP-1 cells without cell toxicity too. Decarine showed anti-inflammatory activity on human colon cells by reducing IL-6 and IL-8 production in TNF-α+IL-1ß-induced Caco-2 cells. These results support the use of Maqian bark as a remedy for enteritis and colitis recorded by Dai medicine in China, and elucidate the major pharmacological compounds in Maqian bark.

Spatholobi Caulis extracts promote angiogenesis in HUVECs in vitro and in zebrafish embryos in vivo via up-regulation of VEGFRs.

ETHNOPHARMACOLOGICAL RELEVANCE: Spatholobi Caulis is a traditional blood-activating and stasis-dispelling herb medicine, which has been used to treat diseases related to blood stasis syndrome (BSS) by inhibiting platelet aggregation, stimulate hematopoiesis, etc. It has been demonstrated that pro-angiogenesis could improve BSS. However, the pro-angiogenic activity of Spatholobi Caulis was not well elucidated AIM OF STUDY: To determine the potential pro-angiogenic activity of Spatholobi Caulis and elucidate its underlying mechanism. The active fractions of Spatholobi Caulis were further screened. MATERIAL AND METHODS: Gelatin precipitation and reversed-phase liquid chromatography (RPLC) were used to purify the methanol extracts of Spatholobi Caulis, respectively. The RPLC was also used to prepare fractions. Total flavonoids of purified methanol extracts of Spatholobi Caulis (PSC) were determined using ultraviolet spectrophotometry. The morphological observation of subintestinal vessel plexus (SIVs) and tyrosine kinase inhibitor II (VRI)-induced intersegmental blood vessels (ISVs) loss in transgenic zebrafish Tg(fli-1a: EGFP)y1 were selected to evaluate the pro-angiogenic activity of PSC in vivo. Cell proliferation by MTT assay and cell migration assay were used to evaluate the pro-angiogenesis effect of PSC in human umbilical vein endothelial cells (HUVECs) in vitro. Both zebrafish and HUVECs were used in screening active fractions of PSC. The mechanism of PSC promoting angiogenesis were studied by real-time PCR in zebrafish and western blotting in HUVECs. RESULTS: Co-treatment PSC dramatically rescued VRI-induced ISVs loss in zebrafish embryos in a dose-dependent manner and 80% of the defective vascular recovered at the concentration of 30µg/ml compared with VRI-only group. PSC also concentration-dependently increased average sprouting number and diameter of SIVs in zebrafish embryo. Real-time PCR assay proved that PSC significantly restored the down regulation of VEGFRs including Flt-1, Kdr and Kdrl induced by VRI in zebrafish (P<0.001). Furthermore, PSC not only promoted proliferation and migration of normal HUVECs but also ameliorated VRI-induced HUVECs cytotoxicity. Western blotting assay showed that co-treatment of PSC increased the expression of VEGFRs and phosphorylation of MAPKs which decreased by VRI treatment. In addition, quality evaluation experiments showed that the content of total flavonoids of PSC reached 56.36% and the main pro-angiogenic fractions of PSC were F3, F4 and F5 both in zebrafish and HUVECs. CONCLUSIONS: Our data demonstrated that PSC presented pro-angiogenic activity both in zebrafish and HUVECs, and principal pro-angiogenic active components were likely flavonoids. Thus, the current study provided evidence for the clinical usage of Spatholobi Caulis in promoting blood circulation and removing stasis in traditional Chinese medicine (TCM).

Research on dissolution rule of main active ingredients in compatibility process of Compound Xuezhining / 中草药

Objective To study the content change of six main active ingredients (orange diclofenac, rubrofusarin-6-O-β-D- gentiobioside, emodin, stilbene glucoside, nuciferine, and quercetin) in the aqueous extract under different compatibility conditions of Compound Xuezhining (CX), using Kohonen maps model for data analysis of traditional Chinese medicine with obvious change in content, to further explore how the compatibility of TCM affect the dissolution characteristic of ingredient. Methods Through different combinations of different herbs in CX, the solubility of the six active ingredients was investigated by UPLC method before and after different compatibility modes; Analyzed the data and chosen Nelumbinis Folium which has significant quantity changes after compatibility as the object of research. The impacts of different compatibility modes on seven chemical compositions of Nelumbinis Folium were analyzed by supervised Kohonen maps model. Results The content change of six active ingredients after compatibility were different: the contents of nuciferine, orange diclofenac and emodin were mainly decreased, the content of stilbene glucoside was decreased, the content of quercetin was increased, and the content of rubrofusarin-6-O-β-D-gentiobioside showed no obvious regularity; When combined with Fructus Crataegi Pinnatifidae, seven chemical constituents in Nelumbinis Folium changed greatly and most of which were positively affected. Conclusion The effects of different compatibility modes on the solubility of active components are different in Chinese herbal formulae CX, and the content of chemical constituents in Nelumbinis Folium are more influential when combined with Fructus Crataegi Pinnatifidae.