A 26-week repeated dose toxicity evaluation of sporoderm-removed Ganoderma lucidum spores in rats.

Ganoderma lucidum spore powder is a traditional Chinese medicine with a variety of health benefits. Sporoderm-removed Ganoderma lucidum spores (RGLS) can be more effectively absorbed and utilized by the body. Due to the extensive clinical application and lack of long-term (>30 days) safety evaluation of RGLS, it is necessary to evaluate its repeated dose toxicity during a longer administration period. Here, we conducted a 26-week repeated dose toxicity test of RGLS in Sprague‒Dawley (SD) rats. The male and female rats were orally administered RGLS at doses of 0, 0.4, 1.2, and 4.0 g/kg once daily for a period of 26 weeks. The safety profile of RGLS was assessed through in vivo observations of survival, body weight, and food consumption; hematological, biochemical, and urine analyses; immunotoxicity assays; and histopathological examinations. The results showed that no significant systemic toxicity was observed following 26 weeks of repeated RGLS administration. Our data showed a no-observed adverse effect level (NOAEL) of 4.0 g/kg, which is approximately 20 times higher than the human equivalent dose. Our results support that RGLS can be considered a safe medicinal or food product that can be added to a healthy diet.

Intestinal microbiota analyses of five economic fishery resources in the South China Sea.

The investigation of intestinal microbiota can provide evidence for revealing the growth and development regulation, feeding habits, environmental adaptability and pollutant indication of marine organisms. To data, the intestinal microbiota of marine organisms in the South China Sea is relatively lacking. To supplement these information, we sequenced intestinal microbiota from five fishery resources (including Auxis rochei, A. thazard, Symplectoteuthis oualaniensis, Thunnus albacores, and Coryphaena equiselis) in the South China Sea using high-throughput Illumina sequencing technology. After filtering, a total of 18,706,729 reads were finally produced and then clustered into OTUs. The mean number of OTUs detected in A. rochei, A. thazard, C. equiselis, S. oualaniensis, and T. albacores was 127, 137, 52, 136, and 142, respectively. Although the Actinobacteria, Bacteroidetes, Cyanobacteria, Deferribacteres, Firmicutes, Proteobacteria, Spirochaetes, Tenericutes, [Thermi], and unclassified_Bacteria were the most abundant in the five species, Photobacterium is the most abundant microbiota. Meanwhile, intestinal microbiota showed species- and sampling sites- specificity, thus only 84 microbiota species were common to all species. Additionally, the potential functions of OTUs in the five species is mainly involved in the synthesis and metabolism of carbohydrate, amino acid, fatty acid and vitamin. This study can provide basic data for clarifying the diversity and species- specificity of intestinal microbiota of five species in the South China Sea, and help to improve the intestinal microbiota database of marine organisms.

Traditional Chinese medicine manipulative reduction combined with percutaneous vertebroplasty for treating type III Kummell's disease: A case report.

BACKGROUND: A patient with type III Kummell's disease had a ruptured posterior cortex of the fractured vertebral body, which caused spinal cord compression. An open surgery was considered the best choice of operation. However, the patient and her family refused open surgery and instead demanded a minimally invasive surgical treatment such as percutaneous vertebroplasty (PVP). After preoperative discussion, we finally adopted the novel therapy of traditional Chinese medicine manipulative reduction (TCMMR) combined with PVP. CASE SUMMARY: A patient with type III Kummell's disease exhibiting bone block-induced spinal cord compression was admitted to our hospital. She suffered from a variety of medical disorders but refused open surgery, and instead asked for PVP surgery. TCMMR, in parallel with PVP, was used to restore the height of the compressed vertebral body and reduce the symptoms of spinal cord compression by the bone block in order to strengthen the vertebral body and prevent further collapse. The surgery was very successful. The height of the compressed vertebra was restored, and the symptom of spinal cord compression by bone block was reduced successfully via TCMMR. The fractured vertebra was solidified by the PVP. The pain visual analog score declined from preoperative 7 scores to postoperative 2 scores, and the Frankel spinal cord scale increased from preoperative D degree to postoperative E degree. CONCLUSION: The new method has advantages in treating patients with type III Kummell's disease who cannot be treated with open surgery.

Promotion of angiogenesis in vitro by Astragalus polysaccharide via activation of TLR4 signaling pathway.

During the implantation of functional tissue-engineered constructs for treating bone defects, a functional vascular network is critical for the survival of the construct. One strategy to achieve rapid angiogenesis for this application is the co-culture of outgrowth endothelial cells (OECs) and primary human osteoblasts (POBs) within a scaffold prior to implantation. In the present study, we aim to investigate whether Astragalus polysaccharide (APS) promotes angiogenesis or vascularization via the TLR4 signaling pathway in a co-culture of OECs and POBs. The co-cultures were treated with various concentrations of APS for 24 h and, subsequently, another 7 days, followed by CD31 staining and analysis of micro-vessel-formation areas using software. Additionally, APS (0.4 mg/ml for 24 h) was added to monocultures of OECs or POBs for evaluating proliferation, apoptosis, angiogenesis, osteogenesis, TLR4 signaling pathway, and inflammatory cytokine release. We found that APS promoted angiogenesis in the co-culture at the optimal concentration of 0.4 mg/ml. TLR4 activation by APS up-regulated the expression level of TLR4/MyD88 and enhanced angiogenesis and osteogenesis in monocultures of OECs and POBs. The levels of E-selectin adhesion molecules, three cytokines (IL-6, TNF-α, and IFN-γ), and VEGF and PDGF-BB, which can induce angiogenesis, increased significantly (p < .05) following APS treatment. Therefore, APS appears to promote angiogenesis and ossification in the co-culture system via the TLR4 signaling pathway. PRACTICAL APPLICATIONS: This study demonstrates that APS may promote angiogenesis and osteocyte proliferation in OEC and POB co-culture systems through the MyD88-dependent TLR4 signaling pathway. APS might represent a potential therapeutic strategy in tissue-engineered bone implantation for the treatment of large bone defects; additionally, it has the advantage of safety, as it exhibits low or no side effects. In the future, it is expected to be used in vitro for the construction of tissue-engineered bone and in vivo after implantation in patients with bone defects for promoting rapid vascularization and ossification of tissue-engineered bone and early fusion with the recipient's bone. In addition, as a food additive, Astragalus membranaceus can be used as a tonic material in patients recovering from a fracture for promoting blood-vessel formation at the fracture site and fracture recovery. Combining traditional Chinese medicine with tissue engineering can provide further strategies for promoting the development of regenerative medicine.

Efficacy of compound traditional Chinese medicine oil in alleviating regorafenib-induced hand-foot skin reaction.

Objective: Regofinib is a novel, oral, anticancer target which greatly improves survival of patients with colorectal cancer. However, it causes 47%-71% patients occur hand-foot skin reaction (HFSR), which affects the quality of life (QOL) or prognosis of patients. However targeted and effective methods are rare. The study attempted to test the efficacy of the compound traditional Chinese medicine (CTCM) oil in relieving HFSR and improving the QOL. Methods: The present study was conducted in 85 patients with metastatic colorectal cancer presenting with HFSR from July 2019 to June 2020. These patients were divided into the control group (n = 42) and the intervention group (n = 43). The HFSR in the control group was managed using traditional methods, whereas that in the intervention group was treated with a CTCM oil comprising five selected herbs. This oil functions by regulating the vascular endothelial growth factor receptor, inflammatory factors, and immune status, and it was provided to the patients to apply externally twice a day. HFSR remission and QOL were evaluated in both groups after 2 weeks of intervention. Results: The post-intervention HFSR remission rate was significantly higher in the intervention group than in the control group (65.1% vs. 16.7%) (P < 0.01). The overall QOL, physical function, social function, and emotional function of the patients in the intervention group were significantly improved (P < 0.05), whereas the pain symptoms were more significantly improved (P < 0.01). Conclusions: The CTCM oil can effectively alleviate HFSR and improve the QOL of patients using regorafenib.

Dual-Directional Regulation of Belamcanda chinensis Extract on Ovalbumin-Induced Asthma in Guinea Pigs of Different Sexes Based on Serum Metabolomics.

To explore the potential biomarkers and metabolic pathways underlying the anti-asthma effects of Belamcanda chinensis extract, this study established an ovalbumin-induced allergic bronchial asthma model in guinea pigs. Sixty guinea pigs were randomly divided into the blank control group, model control group, Belamcanda chinensis extract groups (0.8 g/kg, 1.2 g/kg, 1.6 g/kg, respectively), and dexamethasone acetate tablet group (0.5 mg/kg). Starting on the 22nd day, the drugs were administered by gavage for seven consecutive days. Serum and bronchoalveolar lavage fluid (BALF) were collected. The levels of IL-4 and IgE in the serum and IFN-γ and TNF-α in the BALF were detected by ELISA. UPLC-MS was combined with multivariate statistical analyses, including partial least squares discriminant analysis (PLS-DA), and differential metabolites between groups were identified. Metabolic pathway analysis was performed by querying the KEGG (Kyoto Encyclopedia of Genes and Genomes) online database. Female and male guinea pigs were analyzed. The results showed that compared with the model group, the IgE and IL-4 serum levels were significantly decreased in the 1.6 g/kg group, and the IFN-γ level in the BALF was significantly increased. The TNF-α level was significantly decreased in the 1.2 g/kg and 1.6 g/kg groups. There were 39 common differential metabolites among females and males, and 37 differential metabolites showed opposite regulatory trends in the serum of guinea pigs with asthma and after treatment, mainly involving 17 metabolic pathways, such as pantothenate and CoA biosynthesis, the arachidonic acid mechanism, and glycerophospholipid metabolism. Belamcanda chinensis extract improved OVA-induced asthma, as determined based on immune mechanisms, inflammation, nerve metabolism, and energy metabolism. The serum levels of metabolites produced by the model animals exhibited distinct sex-specific differences, and the treatment effect of Belamcanda chinensis extract also showed sex-specific differences and bidirectional regulation.

Exploring the molecular mechanism of action of Polygonum capitatum Buch-Ham. ex D. Don for the treatment of bacterial prostatitis based on network pharmacology and experimental verification.

ETHNOPHARMACOLOGY RELEVANCE: Polygonum capitatum Buch-Ham. ex D. Don (CNPC2009), a traditional Miao-national herbal medicine, has been widely used with considerable therapeutic efficacy in the treatment of various urologic disorders including prostatitis. However, the molecular mechanism of action (MOA) remains unclear. AIM OF THE STUDY: In this study, UPLC-Q-Exactive-MS and Network pharmacological methods were used to explore the underlying molecular MOA of Polygonum capitatum Buch-Ham. Ex D. Don (P.capitatum) for the treatment bacterial prostatitis (BP). MATERIALS AND METHODS: The UPLC-Q-Exactive-MS technique was used to identify the chemical components of P. capitatum. Databases such as SwissTargetPrediction, Gene Cards, and OMIM were used to predict the targets of P. capitatum for the treatment of BP. The Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) was used to analyze the protein-protein interaction (PPI) and construct a PPI network, and the Metascape was used for Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. In addition, experimental treatment of Escherichia coli (E.coli)-induced BP was verified. RESULTS: A total of 31 molecular components were identified by UPLC-Q-Exactive-MS. Network pharmacology revealed that P. capitatum may act on the AKT1, PI3K, MTO, EGFR and other targets through active components such as Gallic acid, Quercetin, Luteolin, Protocatechuic Acid, Kaempferol and thereby regulate PI3K-AKT, ErbB, AMPK, HIF-1, and other signaling pathways to intervene in the pathological mechanism of BP. Verification through experimental results showed that compared with the model group, treatment with P. capitatum could significantly inhibit bacterial growth in prostate tissues, lowered the prostate index, down-regulated the levels of inflammatory mediators(IL-1ß, IL-6, and TNF-α) in prostate tissues, and down-regulate the protein expression and mRNA expression levels of AKT and PI3K. CONCLUSION: This study preliminarily revealed the MOA of P. capitatum for treating BP with multiple components, multiple targets, and multiple pathways, especially affecting the PI3K-AKT signaling pathways.

Mechanisms of Dendrobium officinale polysaccharides in repairing gastric mucosal injuries based on mitogen-activated protein kinases (MAPK) signaling pathway.

The present study aimed to investigate the protective effects and molecular mechanisms of Dendrobium officinale polysaccharides on gastric mucosal injuries. Following one week of continuous intragastric administration, a gastric mucosal injury model was established using intragastric administration of anhydrous ethanol. The area of gastric ulcer was measured, the contents of interleukin- 6 (IL-6), epidermal growth factor receptor (EGFR), and thyroid transcription factor 1 (TFF-1) in serum were detected by enzyme linked immunosorbent assay (ELISA), and the expressions of EGFR, TFF-1, IL-6, Raf-2, MAP kinase kinase 1 (MEK1), MEK2, and ERK1 in the gastric tissue were determined utilizing qPCR, Western blotting and immunohistochemistry. Simultaneously, Dendrobium officinale polysaccharides and anhydrous ethanol were added to the gastric mucosal cells (GES1) cultured in vitro, and the protective effects of Dendrobium officinale polysaccharides on cell viability was detected using Cell Counting Kit (CCK)-8. The addition of Dendrobium officinale polysaccharides markedly improved the gastric epithelial defect, inflammatory cell infiltration, and redness and swelling stemmed from gastric mucosal injuries and greatly reduced the area of gastric ulcer. The inhibition rates of gastric ulcer were 48.12 ± 2.98, 42.95 ± 1.52, and 27.96 ± 2.05% in the high, medium, and low concentration Dendrobium officinale polysaccharide groups, respectively. Dendrobium officinale polysaccharides could increase the expressions of EGFR and TFF-1 and decrease the expressions of IL-6, Raf-2, MEK1, MEK2, and ERK1. Dendrobium officinale polysaccharides could reduce the level of inflammatory factors and protect gastric mucosa by inhibiting the expression of MAPK pathway genes and proteins.

New Monoterpenoid Indoles with Osteoclast Activities from Gelsemium elegans.

The well-known toxic medicine Gelsemium elegans is widely and historically used to treat bone fracture and skin ulcers by the folk people of China. Two new monoterpenoid indole alkaloids, gelselegandines D and E, together with the known analogue gelegamine A were isolated from G. elegans. Their structures were elucidated by means of spectroscopic techniques and quantum chemical calculations. All isolated compounds were tested for the effects on RANKL-induced osteoclast formation. Interestingly, gelselegandine E and gelegamine A, respectively, showed significant promoting and inhibitory activities on osteoclastogenesis, while gelselegandine D had no activity under the same concentration. This work suggested the different configurations for the carbons near the C-19/20 oxygen rings of the isolated compounds may be the key active groups on osteoclast formation and provided the evidence for the rationality as the traditional treatment for bone-related diseases of G. elegans.

MnO2 nanosheet-mediated target-binding-induced FRET strategy for multiplexed microRNAs detection and imaging in living cells.

In the regulatory network, miRNAs play a regulatory role in a cooperative or antagonistic manner. Simultaneous accurate detection and imaging of multiplexed miRNAs in living cells are of great significance for miRNA-associated biological research and disease diagnosis and treatment. Herein, a MnO2 nanosheet-mediated target-binding-induced fluorescence resonance energy transfer (FRET) strategy was developed for detection and imaging of multiplexed miRNAs in living cells. Two pairs of DNA probes (P1-AF 488/P1'-Cy3 and P2-AF 488/P2'-AF 594) contained the complementary sequence to target miRNAs (miRNA-373 and miRNA-96) and labelled with different fluorescence dyes were designed. They were adsorbed onto MnO2 nanosheets by physisorption to form DNA/MnO2 nanocomposite probes. When the DNA/MnO2 nanocomposite probes were taken up by cells, the MnO2 nanosheets were reduced by intracellular glutathione, accompanying the release of DNA probe pairs. Then the DNA probe pairs specifically recognized and combined with miRNA-373 and miRNA-96 to form stable duplexes, respectively, bringing labelled fluorophores into close proximity to occur FRET. Based on this, the simultaneous imaging of miRNA-373 and miRNA-96 in MDA-MB-231 and L02 cells was successfully implemented. The results displayed a higher expression level of target miRNAs in MDA-MB-231 cells compared to L02 cells. The changes in expression levels of miRNA-96 induced by anti-miRNA-96 or mimics in MDA-MB-231 cells could also be monitored. In addition, the ratiometric detections of multiplexed miRNAs were achieved by utilizing the DNA probe pairs. The proposed strategy provides an alternative method for simultaneous accurate detection and imaging of multiplexed miRNAs and has potential application in biomedical applications.

Genus Alangium - A review on its traditional uses, phytochemistry and pharmacological activities.

The species from Alangium have been used as folk medicine to treat rheumatism, skin diseases, diabetes by the people of Southeast Asia. Previous phytochemical studies have shown this genus are rich sources of alkaloids, glycosides, and terpenoids, which have attracted considerable attention of many researchers due to their markedly diverse and complex architecture. The crude extracts as well as the monomeric compounds from the title genus possess anti-tumor, anti-inflammatory, antibacterial, anti-oxidant pharmacological activities. Besides, some isolates from Alangium exhibited the effects on skeletal, smooth muscle and the nervous system. As a large genus of medicinal plants, the medicinal value of Alangium has been widely reported, but there is no review that provide a systematic summary towards its chemical constituents and pharmacological activities, to our knowledge. This work aims to present a comprehensive overview on the traditional uses, phytochemistry, and pharmacological activities of medicinal plants in the genus Alangium, and to explore the evidence supporting its ethnopharmacological effectiveness.

[Research progress of Rehmanniae Radix in prevention and treatment of osteoporosis].

Osteoporosis fracture with high disability and mortality is a difficult problem that seriously affects the life quality of individuals. At present, there is still a lack of anti-osteoporosis drugs with clear target and significant efficacy in the clinical practice. Rehmanniae Radix and its prescriptions have significant clinical effects. In this regard, more and more studies have reported the effects and mechanisms of Rehmanniae Radix and its active components, and the certain research outputs have been achieved. In this article, the PubMed, Web of science, China National Knowledge Infrastructure, and Wanfang database were searched to collect and organize the latest research progress of Rehmanniae Radix treatment of osteoporosis in the recent 10 years. We summarized the research dynamics as well as the function indexes and mechanisms of the raw and processed Rehmanniae Radix, active ingredients such as catalpol, aucubin, acteoside and Rehmanniae Radix polysaccharide, and their formulating prescriptions, and then excavated the potential active ingredients, targets and signaling pathways, including the effect on bone marrow mesenchymal stem cells, promoting the osteoblast proliferation and promoting osteogenesis differentiation(increasing alkaline phosphatase, typeⅠ collagen, osteoprotegerin, and osteocalcin and promoting calcium deposits), increasing the bone density, inhibiting the osteoclast quantity and differentiation, promoting the osteoclast apoptosis, and reducing tartrate resistant acid phosphatase and bone resorption pit area to provide the reference and develop new ideas for developing Rehmanniae Radix prescriptions for treatment of osteoporosis and exploring its mechanism.

Dendrobine targeting JNK stress signaling to sensitize chemotoxicity of cisplatin against non-small cell lung cancer cells in vitro and in vivo.

BACKGROUND: Lung cancer is a leading cause of cancer-related death worldwide. Cisplatin-based chemotherapy is the standard treatment for lung cancer, but chemoresistance and adverse effects especially cardiotoxicity limit its efficacy. PURPOSE: The efficacy of combination treatment of dendrobine, a plant alkaloid isolated from Dendrobium nobile, with cisplatin was examined as a possible anti-non-small cell lung cancer strategy. METHODS: The cytotoxicity of dendrobine and cisplatin against A549 lung cancer cells was analyzed by MTT and colony formation assays. Apoptosis was measured by annexin V/PI double staining. Apoptosis-related proteins were assessed by western blotting and qPCR analysis. In vivo efficacy was determined using A549 xenograft in nude mice. JNK and Bim inhibition were achieved by siRNA knockdown and/or chemical inhibition. Cardiotoxicity was assessed by serum creatine phosphokinase activity assay. RESULTS: Dendrobine induced apoptotic cell death through mitochondrial-mediated pathway. Combination treatment of dendrobine with cisplatin showed enhanced cytotoxicity through stimulation of JNK/p38 stress signaling pathways and, consequently, the induction of apoptosis involving pro-apoptotic proteins Bax and Bim. In addition, dendrobine attenuated the body weight reduction and cardiotoxicity induced by cisplatin in nude mice. CONCLUSION: The combination treatment showed enhanced anticancer activity toward non-small cell lung cancer cells without aggravating the cardiotoxic effects of cisplatin suggesting that the combination strategy deserves further investigation for human lung cancer treatment.

Isoforskolin and forskolin attenuate lipopolysaccharide-induced inflammation through TLR4/MyD88/NF-κB cascades in human mononuclear leukocytes.

The principal active component of isoforskolin (ISOF) is from the plant Coleus forskohlii, native to China, which has attracted much attention for its biological effects. We hypothesize that ISOF and forskolin (FSK) pretreatment attenuates inflammation induced by lipopolysaccharide (LPS) related to toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and nuclear factor kappa B (NF-κB) signaling. Mononuclear leukocytes (MLs) from healthy donors' blood samples were separated by using density gradient centrifugation. Protein levels of TLR4, MyD88, and NF-κB were detected using western blot and inflammatory cytokines interleukin (IL) 1ß, IL-2, IL-6, IL-21, IL-23, tumor necrosis factor (TNF) α, and TNF-ß were tested by enzyme-linked immunosorbent assay and Quantibody array in MLs. Our results showed that LPS augmented the protein levels of TLR4, MyD88, and NF-κB in MLs and the production of IL-1ß, IL-2, IL-6, IL-21, IL-23, TNF-α, and TNF-ß in supernatants of MLs. Despite treatment with ISOF and FSK prior to LPS, the protein levels of TLR4, MyD88, NF-κB, IL-1ß, IL-2, IL-6, IL-21, IL-23, TNF-α, and TNF-ß in MLs were apparently decreased. roflumilast (RF) and dexamethasone (DM) had a similar effect on MLs with ISOF and FSK. Our results, for the first time, have shown that ISOF and FSK attenuate inflammation in MLs induced by LPS through down-regulating protein levels of IL-1ß and TNF-α, in which TLR4/MyD88/NF-κB signal pathway could be involved.

Baicalin protects mice against Salmonella typhimurium infection via the modulation of both bacterial virulence and host response.

BACKGROUND: The worsening problems of antibiotic resistance prompt the need for alternative strategies. Baicalin, which is isolated from Scutellaria baicalensisi, has been demonstrated to exhibit anti-inflammatory, anti-virulence and antimicrobial effects. Salmonella typhimurium is an important foodborne pathogenic bacteriaum that causes gastrointestinal disease in humans and many animals. PURPOSE: The aim of this study was to investigate the effects of baicalin on S. typhimurium infection in mice and its possible mechanism in vitro. STUDY DESIGN: To evaluate the effect of baicalin in vivo, mice were orally administered of baicalin, and then were infected by an intragastric administration of S. typhimurium. The minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of baicalin, baicalein, and oroxylin A against S. typhimurium were detected under the guides of the Clinical and Laboratory Standards Institute. In vitro, Caco-2 cells were infected with S. typhimurium in the presence or absence of baicalin, baicalein, and oroxylin A at sub-MICs. METHODS: In the in vivo experiment, the body weight loss, the serum levels of TNFα,  IL-6, and lactic dehydrogenase (LDH), the pathological changes of the caecum and the caecum bacterial burdens were examined. The MICs and MBCs of baicalin, baicalein, and oroxylin A against S. typhimurium were detected by two-fold serial dilutions. In vitro, Caco-2 cells were infected with S. typhimurium, and the invasion capacity, TNFα, nitrate, and LDH were analysed. The transcription levels of Salmonella pathogenicity island 1 virulence associated genes (sopB, sopE, sopE2) of S. typhimurium in the presence of baicalin, baicalein, and oroxylin A were detected by qRT-PCR. RESULTS: Our results showed that baicalin significantly decreased the body weight loss, the serum levels of TNFα,  IL-6, and LDH, and the caecum bacterial burdens of mice challenged with S. typhimurium. Histological examination showed that baicalin decreased the lesion in the caecum of S. typhimurium-infected mice. MICs and MBCs of baicalin, and oroxylin A. against S. typhimurium were > 128 µg/ml. MICs and MBCs of baicalein against S. typhimurium were 64 µg/ml, and > 128 µg/ml, respectively. Pretreatment of Caco-2 cells or S. typhimurium with baicalin, baicalein, and oroxylin A significantly inhibited the invasion of Caco-2 cells by S. typhimurium in a dose-dependent manner. Sub-MICs of baicalin, baicalein, and oroxylin A also significantly decreased the levels of TNFα, nitrate, and LDH from S. typhimurium-infected Caco-2 cells. Moreover, the transcription levels of sopB, sopE, and sopE2 were significantly suppressed by baicalin, baicalein, and oroxylin A. CONCLUSIONS: These results demonstrated that baicalin is a promising agent for the prevention of S. typhimurium infection via the modulation of both bacterial virulence and host response.

[Analysis of alcohol extract and water extract of Polygonum capitatum by UPLC-Q-TOF-MS].

In this study, we used Ultra Performance Liquid Chromatography-Time-of-Flight Mass Spectrometry(UPLC-TOF-MS)to identify the chemical constituents in both ethanol and water extract of Polygonum capitatum. A Waters ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7 µm) was used for separation. The mobile phase was consisted of(A) 0.10% formic acid in water and(B)0.10% formic acid in acetonitrile, and the flow rate was 0.35 mL•min⁻¹. ESI source in negative ion mode was used for MS detection. Structural identification was carried out according to the accurate mass and matching with database. The results showed that flavonoids, polyphenols and lignans were the main components in both extracts. However, the chemical compositions of both extracts were different, e.g. there are less hydrolyzable tannins, loss of ellagic acid and more anthocyanins in ethanol extract. In a conclusion, this study provides an important scientific basis for identifying the active ingredients in P. capitatum, which also help to reveal the pharmacological effect of P. capitatum.

Phenylboronic acid modified solid-phase extraction column: Preparation, characterization, and application to the analysis of amino acids in sepia capsule by removing the maltose.

Maltose, a common auxiliary material of pharmaceutical preparation, may disturb the analysis of total amino acids in sepia capsule by aldolization. Therefore, it is necessary to remove the maltose through a convenient method. In this work, a phenylboronic acid modified solid-phase extraction column has been synthesized and used to remove the maltose. The materials were synthesized by one step "thiol-ene" reaction and the parameters of the column such as absorption capacity, recovery, and absorption specificity have been investigated. The results showed the column (0.5 cm of length × 0.5 cm of inner diameter) can absorb 4.6 mg maltose with a linear absorption and absorption specificity. Then this technique was applied in the quantification of amino acids in sepia capsule. After the optimization of the method, four kinds of amino acids, which were the most abundant, were quantified by high-performance liquid chromatography with diode array detection. The amounts of the four kinds of amino acids are 1.5∼2 times more than that without the treatment of solid-phase extraction column, which almost overcomes the influence of the maltose. All the results indicate that the phenylboronic acid modified solid-phase extraction column can successfully help to accurately quantify the total amino acids in sepia capsule.

Interference of strength training can obviously increase the effect of moxibustion treatment on patients with knee joints injuries.

OBJECTIVE: Observe the effects of Acupoint moxibustion combined with muscle training in treating the patients with knee joints strain. METHODS: The 36 patients with knee joint strains were divided into the experimental group and control group with the method of random number table, each group including 18 cases. The control group was treated with Acupoint moxibustion, while the experimental groups were treated with Acupoint moxibustion combined with muscle training. Before the treatment and after the treatment lasting 12 weeks, the therapy effects and improvement of the knee-joint muscle force for the 2 groups of patients were respectively evaluated. RESULTS: Through the treatment of 12 weeks, the clinical symptoms of control group were evidently improved than prior-treatment , but the improvement effects of the knee-joint muscle force (the peak torques of bend and stretch respectively were (32.8 ± 8.8) N·m and (35.0 ± 11.2) N·m were not significant (P > 0.05); while the clinical symptoms and knee-joint muscle force of experimental group (the peak torques of bend and stretch respectively were (40.3 ± 9.3) N·m and (42.3 ± 10.6) N·m were evidently improved than prior-treatment, and the improvement range was also evidently better than the control group (P < 0.05). CONCLUSION: The Acupoint moxibustion combined with muscle force training had synergistic effects in treating the patients with elderly knee-joint strain, could further relieve the pain on knee joints, and improve the joint' s movement, such therapy was worthy to promote and apply in clinic.

Scutellarin Reduces Endothelium Dysfunction through the PKG-I Pathway.

Purpose. In this report, we investigated the protective mechanism of scutellarin (SCU) in vitro and in vivo which could be involved in endothelial cGMP-dependent protein kinase (PKG), vasodilator stimulated phosphoprotein (VASP) pathway, and vascular endothelium dysfunction (EtD). Method. Human brain microvascular endothelial cells (HBMECs) with hypoxia reoxygenation (HR) treatment and rats with cerebral ischemia reperfusion (CIR) treatment were applied. Protein and mRNA expression of PKG, VASP, and p-VASP were evaluated by Western blot and RT-PCR methods. Vascular EtD was assessed by using wire myography to determine endothelium-dependent vasorelaxation in isolated rat basilar artery (BA). Result. In cultured HBMECs, SCU (0.1, 1, and 10 µM) increased cell viability, mRNA, protein level, and phosphorylative activity of PKG and VASP against HR injury. In HR model of BA, SCU increased protein level of P-VASP. In rat CIR model, wire myography demonstrated that SCU (45 and 90 mg/kg, i.v.) significantly reduced ischemic size by partially restoring the endothelium dependent vasodilation of BA; PKG inhibitor Rp-8-Br-cGMPS (50 µg/kg, i.v.) reversed this protection of SCU in CIR rats. Conclusion. SCU protects against cerebral vascular EtD through endothelial PKG pathway activation.

[HPLC Fingerprint and Analysis of Component Difference of Crude and Wine Processed Products for Dipsacus asper Root].

OBJECTIVE: To study the HPLC fingerprint of Dipsacus asper root before and after wine processed, and to compare the changes of fingerprint and chemical composition. METHODS: The HPLC analysis was performed on an Agilent C18 (250 mm x 4. 6 mm, 5 jim) column with gradient elution of acetonitrile-0. 05% phosphoric acid at a flow rate of 1. 0 mL/min, and the column temperature was set at 25 °C. RESULTS: The content of chemical composition changed in different degree after processed, the asperosaponin VI content increased significantly, and two new chromatographic peaks were found in the fingerprint. CONCLUSION: This method can be used to reflect the difference of chemical composition of Dipsacus asper root and its wine processed products. It would he an efficient way for qualitative control of Dipsacus asper root.