Clinical practice guidelines for prevention and treatment of postoperative gastrointestinal disorder with Integrated Traditional Chinese and Western Medicine (2023).

Postoperative gastrointestinal disorder (POGD) was a common complication after surgery under anesthesia. Strategies in combination with Traditional Chinese Medicine and Western medicine showed some distinct effects but standardized clinical practice guidelines were not available. Thus, a multidisciplinary expert team from various professional bodies including the Perioperative and Anesthesia Professional Committees of the Chinese Association of Integrative Medicine (CAIM), jointly with Gansu Province Clinical Research Center of Integrative Anesthesiology/Anesthesia and Pain Medical Center of Gansu Provincial Hospital of Traditional Chinese Medicine and WHO Collaborating Center for Guideline Implementation and Knowledge Translation/Chinese Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) Center/Gansu Provincial Center for Medical Guideline Industry Technology/Evidence-based Medicine Center of Lanzhou University, was established to develop evidence-based guidelines. Clinical questions (7 background and 12 clinical questions) were identified through literature reviews and expert consensus meetings. Based on systematic reviews/meta-analyses, evidence quality was analyzed and the advantages and disadvantages of interventional measures were weighed with input from patients' preferences. Finally, 20 recommendations were developed through the Delphi-based consensus meetings. These recommendations included disease definitions, etiologies, pathogenesis, syndrome differentiation, diagnosis, and perioperative prevention and treatment.

Therapeutic role of Wuda granule in gastrointestinal motility disorder through promoting gastrointestinal motility and decreasing inflammatory level.

Introduction: Previous studies indicated that Wuda Granule (WDG) has been applied in the treatment of gastrointestinal motility disorder (GMD), but the effect and underlying mechanisms is yet to be elucidated. This study aimed to explore the mechanism and pharmacological effect of WDG for GMD via network analysis, verification of animal experiments and clinical experiments. Methods: The chemical components of WDG were identified from the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP, http://lsp.nwu.edu.cn/index.php), and the Encyclopedia of Traditional Chinese Medicine (ETCM, http://www.tcmip.cn/ETCM/index.php/Home/Index/) according to oral bioavailability (OB) ≥ 20% and drug-likeness (DL) ≥ 0.10. The targets of WDG compounds were retrieved from the Swiss Target Prediction database (http://www.swisstargetprediction.ch/) and targets related to GMD were retrieved from GeneCards database (https://www.genecards.org/). Network analysis were performed to screen the key active compounds of WDG and its hub targets. Then the pharmacological effect of WDG were verified via vivo experiments in rats and clinical experiments. Results: The results showed that 117 effective active compounds of WDG were screened and 494 targets of WDG compounds targeting GMD were selected. These targets were involved in the biological process of inflammatory regulation and the regulation of gastrointestinal motility. The mechanism was mainly involved in the regulation of PI3K-Akt signaling pathway and Rap1 signaling pathway. In addition, molecular docking analysis suggested that eight key active compounds of WDG may be mainly responsible for the effect of WDG on GMD by targeting HARS, AKT, and PIK3CA, respectively. Animal experiments and clinical trials both suggested that WDG could exert therapeutical effect on GMD via inhibiting inflammation and promoting gastrointestinal motility, it could also improve digestive function of patients with laparoscopic colorectal cancer after surgery. Conclusion: This study was the first to demonstrate that WDG improved GMD mainly via inhibiting inflammatory level and promoting gastrointestinal motility, providing new insights for the understanding of WDG for GMD, inspiration for future research and reference for clinical strategy in terms of the treatment of GMD.

An inorganic-organic-polymeric nanovehicle for targeting delivery of doxorubicin: Rational assembly, pH-stimulus release, and dual hyperthermia/chemotherapy of hepatocellular carcinoma.

Efficiently synergistic therapy of hepatocellular carcinoma (HCC) by chemotherapeutic drug and photothermal agent remains a considerable challenge. Here, we report a nanodrug that integrates specific hepatoma-targeted delivery, pH-triggered drug release, and cooperative photothermal-chemotherapy function. By grafting the easily self-assembled CuS@polydopamine (CuS@PDA) nanocapsulation with polyacrylic acid (PAA), an inorganic-organic-polymeric hybrid nanovehicle was developed as a dual photothermal agent and carrier for loading antitumor drug-doxorubicin (DOX) through electrostatic adsorption and chemical linking antibody against GPC3 commonly overexpressed in HCC, resulting in the nanodrug, CuS@PDA/PAA/DOX/GPC3. The multifunctional nanovehicle had excellent biocompatibility, stability, and high photothermal conversion efficiency, due to the rationally designed binary CuS@PDA photothermal agent. The 72-h accumulative drug release in pH 5.5 tumor microenvironment can reach up to 84%, far higher than 15% measured in pH 7.4 condition. Notably, in contrast to the merely 20% survival rate of H9c2 and HL-7702 cells exposed to free DOX, their viabilities in the nanodrug circumstance can maintain 54% and 66%, respectively, suggesting the abated toxicity to the normal cell lines. When exposed to the hepatoma-targeting nanodrug, the viability of HepG2 cells was found to be 36%, which further drastically declined to 10% plus 808-nm NIR irradiation. Moreover, the nanodrug is potent to cause tumor ablation in HCC-modeled mice, and the therapeutic efficacy can be greatly enhanced under NIR stimulus. Histology analyses reveal that the nanodrug can effectively alleviate the chemical damage to heart and liver, as compared to free DOX. This work thus offers a facile strategy for design of targeting anti-HCC nanodrug toward combined photothermal-chemotherapy.

Comparison for quantification of eight components in Alpinia officinarum Hance by using high-performance liquid chromatography coupled with diode array detector and charged aerosol detector with individual and substitute reference compound.

An efficient HPLC-DAD-CAD method was developed and compared for simultaneous quantification of four flavonoids and four diarylheptanoids in Alpinia officinarum Hance (A. officinarum) using individual and substitute reference compound. All calibration curves for investigated analytes showed good linear regression (R2> 0.9991). The LODs of investigated compounds for DAD and CAD were 0.15-7.92 ng (0.03-1.58 µg/mL) and 2.91-3.95 ng (0.58-0.79 µg/mL), respectively, whereas the LOQs were 0.52-26.39 ng (0.10-5.28 µg/mL) for DAD, and 9.70-13.18 ng (1.94-2.64 µg/mL) for CAD. Recoveries of all analytes, which ranged from 96.58% to 100.06% for DAD, and from 96.29% to 99.61% for CAD, were acceptable. According to the quantitative results, the eight compounds in A. officinarum can be accurately quantified with individual calibration curves by two detectors. In addition, to overcome the bottleneck of shortage of reference standards, diphenylheptane A and galangin, respectively, were selected for direct or calibrated quantitative determination of other diarylheptanoids and flavonoids in A. officinarum. The results showed the contents of eight components in A. officinarum determined by these methods were similar, which suggested that substitute reference compound was suitable for quantification of its analogues.

In silico analysis of the potential mechanism of a preventive Chinese medicine formula on coronavirus disease 2019.

ETHNOPHARMACOLOGICAL RELEVANCE: With the spread of Coronavirus Disease (2019) (COVID-19), combination with traditional Chinese medicine (TCM) has been widely used as a prevention and therapy strategy in China. Xin guan No.1 (XG-1) prescription is a preventive formula recommended by the Hunan Provincial Administration of TCM to prevent the pandemic of COVID-19. AIM OF THE STUDY: To explore the potential preventive mechanisms of XG-1 against COVID-19 in the combination of network pharmacology approach, single-cell RNA expression profiling analysis, molecular docking and retrospective study. MATERIALS AND METHODS: Encyclopedia of Traditional Chinese Medicine (ETCM) database was used to determine the meridian tropism, active components and target genes of XG-1. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis were conducted by R Cluster Profiler package (3.14.3). Single cell RNA sequencing (scRNA-seq) data of human lung (GSE122960) was downloaded from Gene Expression Omnibus (GEO) database and analyzed by R Seurat package (3.1.2). Cytoscape (3.7.2) was used to construct the interaction network. The main ingredients in XG-1 were identified by HPLC- Q-TOF- MS and used for molecular docking with COVID-19 3CL hydrolytic enzyme and angiotensin converting enzyme II (ACE2). A retrospective study of 47 close contact participants from Dongtang Community of Hunan Province was conducted to evaluated the preventive effect of XG-1. RESULTS: According to the network pharmacology analysis, XG-1 formula was closely related to lung-, spleen- and stomach-meridians and include a total of 206 active components and 853 target genes. GO and KEGG pathway enrichment revealed that XG-1 mainly regulated cellular amino acid metabolism process and neuroactive ligand-receptors interaction. The scRNA-seq profiling showed that angiotensin converting enzyme 2 (ACE2) was principally expressed in alveolar type 2 epithelial cells (AT2). 153 genes were up-regulated in AT2 cells expressing ACE2 and 12 genes were obtained by intersecting with XG-1 target genes, of which 3 were related to immunity. Five main chemical ingredients were detected in XG-1 sample by HPLC-Q-TOF-MS. The molecular docking showed that Rutin, Liquiritin and Astragaloside Ⅳ had a good affinity with COVID-19 3CL hydrolytic enzyme and ACE2. Compared with participants who didn't take XG-1, preventive treatment with XG-1gradules resulted in a significant lower rate of testing positive for SARS-CoV-2 nucleic acid (P < 0.0001). CONCLUSION: The present study showed that XG-1 exerts a preventive effect in close contacts against COVID-19. The underlying mechanism may be related to modulate immunity response through multiple components, pathways, and several target genes co-expressed with ACE2. These findings provide preliminary evidences and methodological reference for the potential preventive mechanism of XG-1 against COVID-19.

Characterization of the complete chloroplast genome of an endangered ornamental and medicinal plant Lilium pumilum.

Lilium pumilum DC. is a useful plant species not only for its showy flowers but also for its edible and medicinal values. Here we report on the complete chloroplast genome sequence of L. pumilum. The chloroplast genome is 152,573 bp in size and includes two inverted repeat regions of 52,984 bp, which is separated by a large single-copy region of 82,009 bp and a small single copy region of 17,580 bp. A total of 130 genes were predicted, including 38 tRNA, 8 rRNA, and 84 protein-coding genes. Phylogenetic analysis placed L. pumilum under the family Liliaceae.

Fenugreek attenuates obesity-induced inflammation and improves insulin resistance through downregulation of iRhom2/TACE.

AIMS: We previously reported that fenugreek-derived 4-hydroxyisoleucine ameliorates insulin resistance via regulation of TNF-α converting enzyme (TACE) expression. In the present study, we further investigate the effects and mechanisms of fenugreek on obesity-induced inflammation and insulin signaling in the high-fat diet (HFD)-challenged obese mice. MAIN METHODS: After 12 weeks of HFD intervention, mice were treated with the low or high dosages of fenugreek. Serum levels of glucose, insulin, lipid profile, inflammation cytokines, and adipokines were detected. Macrophage infiltration and adipose tissue morphology were observed. Western blot was conducted to investigate the expressions of inactive rhomboid 2 (iRhom2) and TACE as well as other signaling pathways in subcutaneous adipose tissue. KEY FINDINGS: We showed that fenugreek significantly suppressed body weight gain and fat accumulation in HFD-challenged obese mice. Meanwhile, fasting glucose, insulin, and HOMA-IR in fenugreek-treated mice were remarkably decreased, which were properly explained by fenugreek-induced activation of the insulin receptor signaling pathway. Moreover, the anti-inflammatory properties of fenugreek were shown by the decrease of systemic and local expressions of pro-inflammatory cytokines as well as reduced macrophage infiltration into adipose tissue. Additionally, fenugreek markedly deactivated NF-κB and JNK pathways. Finally, we demonstrated that fenugreek strikingly repressed the transcriptions and expressions of iRhom2 and TACE. SIGNIFICANCE: Fenugreek shows an encouraging and promising property in ameliorating insulin resistance and suppressing inflammation in obesity, which might be realized by fenugreek-mediated inhibition of iRhom2/TACE axis-facilitated TNF-α release from adipocytes.

Chaihu-Longgu-Muli decoction relieves epileptic symptoms by improving autophagy in hippocampal neurons.

ETHNOPHARMACOLOGICAL RELEVANCE: Chaihu-Longgu-Muli decoction (CLMD) is a well-known ancient formula in traditional Chinese medicine (TCM) to relieve disorder, clear away heat, tranquilize the mind and allay excitement. It has been used for the therapy of neuropsychiatric disorders such as epilepsy, dementia, insomnia, anxiety, and depression for several centuries in China. AIM OF THE STUDY: This paper is based on the assumption that the mechanism by which CLMD relieves epileptic symptoms in rats is associated with improving autophagy. Several experimental methods are designed to testify the hypothesis. MATERIALS AND METHODS: The lithium-pilocarpine-induced epilepsy model was established in rats. The seizure frequency was recorded. Morphology and number of autophagosomes in hippocampal dentate gyrus was detected with a transmission electron microscope (TEM). Expression of Beclin-1, microtubule-associated proteins 1A/1B light chain 3 (LC3), and mammalian target of rapamycin (mTOR) in dentate gyrus was measured by immunofluorescence assay, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western-blotting. RESULTS: CLMD could significantly relieve the seizure frequency and improve autophagy in hippocampal dentate gyrus. Meanwhile, the level of Beclin-1 and LC3B decreased significantly, while mTOR increased remarkably after medical intervention. CONCLUSIONS: CLMD could improve autophagy in hippocampal dentate gyrus due to epilepsy, especially at high dose. The mechanism may be related to upregulated expression of mTOR and downregulated expression of Beclin-1 and LC3B.

Comprehensive comparison of the anti-inflammatory activity and chemical consistency of traditional Chinese medicine formula granules with Ge-Gen decoction as a representative sample.

Traditional Chinese medicine formula granules (TCMFGs), an advanced dosage form of traditional Chinese medicine, are entering the market on a large scale. However, little attention has been paid to the simultaneous efficacy assessment and quality control of this advanced dosage form. In this study, a comprehensive comparison of the pharmacological activity and chemical consistency of TCMFGs from different manufacturers was performed. Ge-Gen decoction (GGD) samples were used as the target TCMFG. The in vitro anti-inflammatory effects among different types of GGDs indicate that all of them showed different abilities to reduce the lipopolysaccharide-activated production of nitric oxide, interleukin-1ß (IL-1ß), IL-6 and tumor necrosis factor-α. The results from a dimethylbenzene-induced inflammation model in mice indicated that the nine samples in this study showed significant in vivo anti-inflammatory effects. Qualitative and quantitative analyses were performed by multiwavelength ultra-high-performance liquid chromatography with diode array detection and electrospray ionization with quadrupole time-of-flight-tandem mass spectrometry. To visually interpret the differences in the chemical materials, a scatter plot analysis was performed. According to the scatter plot analysis, nine compounds were evaluated as important contributors to the differences. This is the first report of TCMFGs on the basis of the spectrum-effect consistency.

Anti-fibrotic Effects and Mechanism of Shengmai Injection () on Human Hepatic Stellate Cells LX-2.

OBJECTIVE: To investigate the effects of Shengmai Injection (, SMI) on the proliferation, apoptosis and N-myc downstream-regulated gene 2 (NDRG2, a tumour suppressor gene) expression in varying densities of human hepatic stellate cells LX-2. METHODS: LX-2 cells were cultured in vitro. Then, cells were plated in 96-well plates at an approximate density of 2.5×104 cells/mL and cultured for 48, 72, 96 or 120 h followed by the application of different concentrations of SMI (0.6, 1.2, 2.4, 4.8 or 6 µL/mL). Cell proliferation was measured after an additional 24 or 48 h using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of SMI on different cell growth states (cultured for 48, 72, 96, or 120 h) were observed by light microscopy at 24 h after treatment. When the cells reached 80% conflfluence, apoptosis was detected by flflow cytometry after 24 h. Lastly, LX-2 cells were treated with different concentrations of SMI and extracted with protein lysis buffer. The levels of NDRG2 were measured by Western blot. RESULTS: When the LX-2 cells grew for 48, 72, 96 and 120 h, 4.8 and 6 µL/mL of SMI significantly inhibited cell proliferation at 24 and 48 h after treatment (P<0.05). And 2.4 µL/mL of SMI also inhibited cell proliferation at 24 h after treatment when cell growth for 48 h (P<0.05) and at 48 h after treatment when cell growth for 72, 96 and 120 h (P<0.05). The NDRG2 expression level in the LX-2 cell was significantly increased when treated with SMI at concentrations of 1.2, 2.4, 4.8 or 6 µL/mL (P<0.05). CONCLUSION: The inhibitory effects of SMI on the proliferation of LX-2 cells were related to not only concentration dependent but also cell density. In addition, SMI (2.4, 4.8 and 6 µL/mL) could accelerate apoptosis in LX-2 cells, and the mechanism might be associated with NDRG2 over-expression.

Buyang huanwu decoction combined with BMSCs transplantation promotes recovery after spinal cord injury by rescuing axotomized red nucleus neurons.

ETHNOPHARMACOLOGICAL RELEVANCE: Buyang huanwu decoction (BYHWD) is a classic recipe in traditional Chinese medicine (TCM) to supplement Qi and activate blood. It has been used to recover the neural function after the injury of central nervous system for hundreds of years in China. AIM OF THE STUDY: This study investigated whether Buyang huanwu decoction (BYHWD) combined with bone marrow mesenchymal stem cells (BMSCs) transplantation had synergistic effect on neuroprotection of red nucleus neurons after spinal cord injury (SCI). MATERIALS AND METHODS: Rubrospinal tract (RST) transection model was established and BMSCs were collected. The forelimb locomotor function was recorded using inclined plate test and spontaneous vertical exploration. cAMP level in red nucleus was detected with Enzyme-linked immunosorbent assay (ELISA). Morphology and number of red nucleus neurons was observed using Nissl's staining. Expression of cAMP-response element binding protein (CREB), ras homolog gene family member A (RhoA) and nerve growth factor (NGF) in red nucleus was detected using immunohistochemistry, qRT-PCR and Western-blotting. RESULTS: The combination of BYHWD and BMSCs transplantation could improve the forelimb locomotor function significantly and give the red nucleus somas a better protection. Meanwhile, cAMP level, CREB and NGF increased, while RhoA decreased remarkably in the BYHWD+BMSCs group. CONCLUSIONS: BYHWD combined with BMSCs transplantation had synergistic effect on neuroprotection of red nucleus neurons after SCI; the mechanism may be related to up-regulating cAMP level, activating the cAMP/CREB/RhoA signaling pathway, and promoting expression of NGF.

Anti-fibrotic Effects and Mechanism of Shengmai Injection () on Human Hepatic Stellate Cells LX-2 / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of Shengmai Injection (, SMI) on the proliferation, apoptosis and N-myc downstream-regulated gene 2 (NDRG2, a tumour suppressor gene) expression in varying densities of human hepatic stellate cells LX-2.@*METHODS@#LX-2 cells were cultured in vitro. Then, cells were plated in 96-well plates at an approximate density of 2.5×10 cells/mL and cultured for 48, 72, 96 or 120 h followed by the application of different concentrations of SMI (0.6, 1.2, 2.4, 4.8 or 6 μL/mL). Cell proliferation was measured after an additional 24 or 48 h using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of SMI on different cell growth states (cultured for 48, 72, 96, or 120 h) were observed by light microscopy at 24 h after treatment. When the cells reached 80% conflfluence, apoptosis was detected by flflow cytometry after 24 h. Lastly, LX-2 cells were treated with different concentrations of SMI and extracted with protein lysis buffer. The levels of NDRG2 were measured by Western blot.@*RESULTS@#When the LX-2 cells grew for 48, 72, 96 and 120 h, 4.8 and 6 μL/mL of SMI significantly inhibited cell proliferation at 24 and 48 h after treatment (P<0.05). And 2.4 μL/mL of SMI also inhibited cell proliferation at 24 h after treatment when cell growth for 48 h (P<0.05) and at 48 h after treatment when cell growth for 72, 96 and 120 h (P<0.05). The NDRG2 expression level in the LX-2 cell was significantly increased when treated with SMI at concentrations of 1.2, 2.4, 4.8 or 6 μL/mL (P<0.05).@*CONCLUSION@#The inhibitory effects of SMI on the proliferation of LX-2 cells were related to not only concentration dependent but also cell density. In addition, SMI (2.4, 4.8 and 6 μL/mL) could accelerate apoptosis in LX-2 cells, and the mechanism might be associated with NDRG2 over-expression.

Screening and Identification of Antioxidants from Ophiocordyceps xuefengensis (Ascomycetes) by Using DPPH-HPLC-DAD-Q-TOF-MS/MS.

Ophiocordyceps xuefengensis has recently been identified as a sister taxon of O. sinensis and has long been recognized as a folk tonic, food, or invigorant in South China. This study had 3 aims: First, we analyzed the antioxidant activities of 3 kinds of caterpillar fungi; the antioxidative ability of the various caterpillar fungi were in the order of O. xuefengensis > Cordyceps militaris > O. sinensis. Second, we further identified the antioxidant compounds in O. xuefengensis by using various methods: multiwavelength DPPH high-performance liquid chromatography with diode array detection and quadrupole time-of-flight mass spectrometry. In this way, we found and identified 30 compounds that possessed potential antioxidant activity. Among these compounds, 23 compounds were determined to be composed of nucleosides, nucleotides, and amino acids. To our knowledge, several compounds in O. xuefengensis are reported here for the first time. Third, we compared the antioxidant markers in 3 caterpillar fungi. Results showed that the caterpillar fungi share some common antioxidants, while each also had its own characteristic compounds that correspond to its antioxidant capacity. This is to our knowledge the first report of the screening and identification of antioxidants in O. xuefengensis, and these results might provide beneficial contributions to understandings of O. xuefengensis on the basis of its chemical compounds and antioxidant activity.

Licorice root extract and magnesium isoglycyrrhizinate protect against triptolide-induced hepatotoxicity via up-regulation of the Nrf2 pathway.

Triptolide, the predominant biologically active component of the Chinese herb Tripterygium wilfordii Hook f., possesses numerous pharmacological activities, including anti-inflammatory, anti-fertility, anti-neoplastic, and immunosuppressive effects. However, toxicity and severe adverse effects, particularly hepatotoxicity, limit the clinical application of triptolide. Licorice root extract contains various bioactive compounds and is potent hepatoprotective. Magnesium isoglycyrrhizinate, a magnesium salt of the 18α-glycyrrhizic acid stereoisomer of glycyrrhizic acid, is used clinically in China to treat chronic viral hepatitis and acute drug-induced liver injury. The aim of this study was to investigate the role of the factor erythroid 2-related factor 2 pathway in the protective effects of LE and MIG against triptolide-induced hepatotoxicity. Hepatotoxicity models were established in L-02 cells and rats using triptolide, and the protective effects of LE and MIG were investigated in vitro and in vivo, respectively. LE and MIG significantly protected against triptolide-induced cytotoxicity. Additionally, triptolide decreased the mRNA and protein levels of Nrf2 and down-regulated Nrf2 target genes, including UGT1A, BSEP, and MRP2, while pretreatment with LE and MIG reversed these effects. Finally, Nrf2-involved antioxidant responses were activated in the presence of LE and MIG.

The pharmacological properties of Ophiocordyceps xuefengensis revealed by transcriptome analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: The Yao ethnic group in Xuefeng Mountains area have used Xuefeng cordyceps, the caterpillar-fungus complex of Ophiocordyceps xuefengensis, for treating a variety of diseases for long. Just like some other cordyceps, O. xuefengensis, which is identified as the sister taxon of O. sinensis in 2013, also seems to have broad pharmacological properties, not only enhancing human immunity, anti-bacteria, anti-virus, but also anti-tumor. However, investigation of the medicinal fugal species O. xuefengensis can be found only in few literature records since its pharmacological and therapeutic use is mainly in traditional Yao communities by local healers. AIM OF THE STUDY: The aim of this study is to collect samples of Xuefeng cordyceps and isolate the strain of O. xuefengensis, to determine bioactive components and evaluate the anti-tumor activity, to obtain the gene expression profile of O. xuefengensis and reveal its pharmacological properties by de novo transcriptome analysis. Accordingly, we attempt to provide information and give a comprehensive understanding of this mysterious medicinal fugal species from traditional Yao communities of China. MATERIAL AND METHODS: Bioactive components were determined with HPLC-DAD-Q-TOF-MS technology; in vitro anti-tumor activity against 6 cell lines was evaluated using standard MTT assay; transcriptome analysis was done by de novo sequencing; unique genes were functionally profiled basing on Gene Ontology Database and the targeted genes were examined by blast. RESULTS: Trace cordycepin, an anti-tumor agent, was detected in O. xuefengensis water extract. To some extent, the raw water extract of O. xuefengensis showed in vitro anti-tumor activity, against A549, HepG2, MCF-7, PC-3 and Raji cell lines. A total of 94,858 transcripts and 49,001 unique genes were obtained, amongst, 43.4% unique genes were matched with those of O. sinensis. Not all supposed genes related to cordycepin biosynthetic pathways were found by transcriptome analysis. CONCLUSION: According to the gene expression profile, O. xuefengensis is very close to medicinal fungus O. sinensis. Raw water extract of O. xuefengensis, to a certain degree, could inhibit the growth of tumor cells, indicating that this fungus could be a new resource for the exploration of anti-tumor drug.

A new cordycepin-producing caterpillar fungus Ophiocordyceps xuefengensis with artificial infection to the host, cultivation of mycelia and stromata.

Caterpillar fungi have numerous pharmacological and therapeutic applications in traditional medicine, due to a variety of active chemical constituents, such as cordycepin and adenosine. It is imperative to discover new resource for artificial cultivation and biometabolite production since the traditional natural species are endangered. In this study, a new strain HACM 001 was isolated and identified as Ophiocordyceps xuefengensis by rDNA-ITS sequencing. This strain showed the potential of artificial infection to caterpillar larvae leading to mummification, as well as fermentation mycelia in liquid culture and cultivation stromata in solid medium. Eight nucleosides and nucleobases, especially cordycepin and adenosine, were determined and analyzed with HPLC-DAD-Q-TOF-MS/MS technology. Cordycepin was detected in all forms of present O. xuefengensis strain at different contents, among which the highest content (37.1 µg/g) appeared in the stromata cultivated on solid medium. The content of adenosine in mycelia and stromata, respectively, reached 1155 µg/g and 1470 µg/g. Therefore, O. xuefengensis might be an alternative source for obtaining artificial fungus-caterpillar-larvae complex and producing cordycepin and adenosine.

Anti-fibrotic effects of Salvia miltiorrhiza and Ligustrazine Injection on LX-2 cells involved with increased N-myc downstream-regulated gene 2 expression.

OBJECTIVE: To investigate the effects of Salvia miltiorrhiza and Ligustrazine Injection (SML) on proliferation and apoptosis of human hepatic stellate cell LX-2 and the expression of N-myc downstreamregulated gene 2 (NDRG2, a tumor suppressor gene). METHODS: HSCs from the LX-2 cell line were cultured in vitro. The proliferative state of different initial LX-2 cell numbers was measured using a 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. LX-2 cells were plated in 96-well plates at an approximate density of 2.50×104 cells/mL and cultured for 24 h followed by the application of different concentrations of SML (1, 2, 4 and 8 µL/mL). Cell proliferation was measured using the MTT assay at 24 and 48 h. Apoptosis was detected by flow cytometry at 24 h. LX-2 cells were treated with different concentrations of SML and extracted with protein lysis buffer. The levels of NDRG2 and ß-catenin were measured by Western blot. RESULTS: With the exception of the 1 and 2 µL/mL concentrations, 4 and 8 µL/mL SML inhibited cell proliferation in a concentration-dependent manner at 24 and 48 h (P<0.05). With the exception of the 1 and 2 µL/mL concentrations, the NDRG2 expression level was greatly increased in a concentration-dependent manner. However, the level of ß-catenin was unaffected. CONCLUSION: SML inhibit LX-2 cell proliferation in a concentration-dependent manner, and the mechanism may be associated with NDRG2 over-expression.

Study of Evodia hot compress plus electro-acupuncture in patients who have undergone abdominal surgery.

BACKGROUND: The importance of postoperative gastrointestinal function recovery is being increasingly recognized. In this multi-center randomized controlled study, we evaluated the efficacy and safety of Evodia hot compress (ECS) plus electro-acupuncture (EA) for patients who developed postoperative gastrointestinal tract dysfunction after abdominal surgery. METHODS: 1009 patients who developed postoperative gastrointestinal tract dysfunction after abdominal surgery were enrolled. All patients received conventional therapies for 7 days from the 1st postoperative day and were randomly assigned to receive coarse salt hot compress, Evodia hot compress or Evodia hot compress plus electro-acupuncture twice a day for 7 days. RESULTS: The mean time to first flatus and to first bowel sounds was comparable among the four groups (P>0.05). The control group had a significantly shorter time to defecation compared with patients receiving coarse salt hot compress, Evodia hot compress or Evodia hot compress plus electro-acupuncture (P<0.05). In patients undergoing open hepatectomy, the time to first defecation was the shortest in those who received Evodia hot compress plus electro-acupuncture (89.3±25.5 h), which was significantly different from that of controls(134±31.1 h), those who received coarse salt hot compress (106.7±36.4 h) and those who received Evodia hot compress (109.9±42.1 h) (P<0.05) in patients undergoingopen cholecystectomy, the time to first defecation was the shortest in those who received Evodia hot compress (73.1± 24.7), which was significantly different from that of controls (77.8±29.7), those who received coarse salt hot compress 90.5±30.2) and those who received Evodia hot compress plus electro-acupunctur (83.9±34.0). CONCLUSION: Evodia hot compress plus electro-acupuncture confers benefit in postoperative recovery of gastrointestinal function of patients who have undergone abdominal surgery and it is overall safe to use. TRIAL REGISTRATION: Chinese Clinical Trial RegistryChiCTR-TRC-09000527.

Smilax china L. rhizome extract inhibits nuclear factor-κB and induces apoptosis in ovarian cancer cells.

OBJECTIVE: To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome (SCR) on ovarian cancer cells. METHODS: Ovarian cancer cells A2780 were treated with different concentrations of SCR extract (SCRE), and compared with controls. Effects on cell growth were evaluated by cell counting kit-8 (CCK-8) assay; proliferation effects by EdU incorporation assay; cell cycle by propidium iodide staining; apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide; cellular distribution of nuclear factor-κB (NF-κB) by immunofluorescence; protein levels of NF-κB, caspase-3, poly-adenosine diphosphate (ADP)-ribose polymerase (PARP), Bcl-2-associated X protein (Bax), cellular inhibitor of apoptosis (cIAP)-1, anti-X-linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma-extra large (Bcl-XL), B-cell lymphoma-2 (Bcl-2) and AKT by Western blotting; and effects of SCRE combined with cisplatin or adriamycin on A2780 cells by CCK-8 assay. RESULTS: SCRE suppressed A2780 cell proliferation in a dose-dependent manner (P<0.05,P<0.01), arrested cells in G2/M phase and induced apoptosis by activating caspase-3, PARP and Bax. SCRE treatment also correlated with inhibition of NF-κB and downregulation of Bcl-2, Bcl-XL, cIAP-1, XIAP and AKT. SCRE can promote chemosensitivity to cisplatin and adriamycin in A2780 cells (P<0.01). CONCLUSION: SCR effectively inhibits NF-κB, induces apoptosis and reduces chemoresistance to cisplatin and adriamycin in ovarian cancer cells, which might be its molecular basis for treating ovarian cancer.

[Effect of Trigonella foenum-graecum 4-hydroxyisoleucine on high-glucose induced insulin resistance in 3T3-L1 adipocytes of mice].

OBJECTIVE: To investigate the effect of 4-hydroxyisoleucine (4-HIL), an active component of Trigonella Foenum-graecum L. on high glucose induced insulin resistance (IR) in 3T3-L1 adipocytes, and to explore underlying molecular mechanisms. METHODS: 3T3-L1 adipocytes were treated with 25 mmol/L glucose and 0.6 nmol/L insulin to induce IR. They were intervened by different concentrations of 4-HIL (at 5, 10, and 20 micromol/L). [3H]-Deoxy-D-glucose up-taking method was used to detect the glucose uptake. The mRNA expression of cellular tumor necrosis factor-alpha (TNF-alpha) was detected by polymerase chain reaction (PCR). The content of TNF-alpha in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Palmitic acid (PA) acted as the control. RESULTS: After intervened by 25 mmol/L glucose and 0.6 nmol/L insulin for 18 h, the insulin-stimulated glucose transportation in 3T3-L1 adipocytes was inhibited by 63%. The mRNA expression of cellular TNF-alpha in adipocytes significantly increased, when compared with that in normal adipocytes (P < 0.05). The level of TNF-alpha secreted in the culture supernatant was increased by 70 pg/mL (P < 0.05). Similar changes occurred in the PA group. After exposure to 4-HIL (5, 10, or 20 micromol/L) for 24 h, the glucose transportation was increased by 35%, 50%, and 60%, respectively. PCR results showed that along with increasing 4-HIL concentrations, the mRNA expression of cellular TNF-alpha showed a decreasing trend, showing statistical difference when compared with the model group and the PA group (P < 0.05). Compared with the model group, the TNF-alpha level in the supernatant was respectively reduced by 10 pg/mL, 18 pg/mL, and 39 pg/mL after intervention (P < 0.05). CONCLUSION: 4-HIL could remarkably improve high glucose-induced IR in 3T3-L1 adipocytes. Meanwhile, 4-HIL could inhibit the secretion of TNF-alpha.