Adventitious root culture of Lessertia frutescens for the production of triterpenoid saponins and polysaccharides.

Lessertia frutescens is a perennial shrub of commercial importance in South Africa, but the scarcity of plant resources has limited current product production. In this study, to provide an alternative approach for obtaining L. frutescens material, adventitious roots (ARs) were induced from sterilized seedlings and cultured in a suspension culture system. During this process, selection tests were conducted to find a suitable auxin and its concentration for AR induction and a suitable basal medium for AR growth and metabolite accumulation; a kinetic study was then performed to constructure kinetic models. The results showed that compared to other auxins and concentrations, indole-3-butyric acid at 3 mg/L was suitable for increasing the number and length of ARs during AR induction. In AR suspension culture, Schenk and Hildebrandt (SH) was better than other basal media, and the maximum AR fresh (86.9 g/L) or dry weight (5.5 g/L), total triterpenoid saponin (92.6 mg/g DW), and polysaccharide (114.7 mg/g DW) contents were determined in the 1.5×SH medium. In addition, AR biomass and metabolite contents reached the maximum on day 42. The kinetic models for AR growth and triterpenoid and polysaccharide production were constructed, providing the basis for further optimization of culture conditions and large-scale culture.

Evaluation of the "Relative Ordered Structure of Hericium erinaceus Polysaccharide" from Different Origins: Based on Similarity and Dissimilarity.

Polysaccharides are organic compounds widely distributed in nature, but structural order and disorder remain a formidable problem. In this study, based on the theoretical framework of the "relative ordered structure of polysaccharide" proposed in our previous work, the structural order of Hericium erinaceus polysaccharides from different regions was evaluated by FT-IR, methylation analysis, and 1H NMR spectroscopy combined with chemometric methods. The results of principal component analysis and heatmap cluster analysis revealed that 18-subfractions exhibit four different structural types with representative glycoside linkage types: fucogalactoglucan, glucofucogalactan, fucoglucan, and glucan. The main chain of heteroglucans often consists of ß-(1 → 6)-Glcp, ß-(1 → 4)-Glcp, and ß-(1 → 3)-Glcp residues, which are predominantly substituted at the O-3 and O-6 positions. The main chain structure of heterogalactans is α-(1 → 6)-Galp residues, which may be replaced by Fucp and Galp residues at O-2. Overall, our findings demonstrate the validity of the "relative ordered structure of polysaccharide" in Hericium erectus polysaccharides and simplify the complexity of polysaccharide structures.

Exploring the partial degradation of polysaccharides: Structure, mechanism, bioactivities, and perspectives.

Polysaccharides are promising biomolecules with lowtoxicity and diverse bioactivities in food processing and clinical drug development. However, an essential prerequisite for their applications is the fine structure characterization. Due to the complexity of polysaccharide structure, partial degradation is a powerful tool for fine structure analysis, which can effectively provide valid information on the structure of backbone and branching glycosidic fragments of complex polysaccharides. This review aims to conclude current methods of partial degradation employed for polysaccharide structural characterization, discuss the molecular mechanisms, and describe the molecular structure and solution properties of degraded polysaccharides. In addition, the effects of polysaccharide degradation on the conformational relationships between the molecular structure and bioactivities, such as antioxidant, antitumor, and immunomodulatory activities, are also discussed. Finally, we summarize the prospects and current challenges for the partial degradation of polysaccharides. This review will be of great value for the scientific elucidation of polysaccharide fine structures and potential applications.

Improving Flavonoid Accumulation of Bioreactor-Cultured Adventitious Roots in Oplopanax elatus Using Yeast Extract.

Oplopanax elatus is an endangered medicinal plant, and adventitious root (AR) culture is an effective way to obtain its raw materials. Yeast extract (YE) is a lower-price elicitor and can efficiently promote metabolite synthesis. In this study, the bioreactor-cultured O. elatus ARs were treated with YE in a suspension culture system to investigate the elicitation effect of YE on flavonoid accumulation, serving for further industrial production. Among YE concentrations (25-250 mg/L), 100 mg/L YE was the most suitable for increasing the flavonoid accumulation. The ARs with various ages (35-, 40-, and 45-day-old) responded differently to YE stimulation, where the highest flavonoid accumulation was found when 35-day-old ARs were treated with 100 mg/L YE. After YE treatment, the flavonoid content increased, peaked at 4 days, and then decreased. By comparison, the flavonoid content and antioxidant activities in the YE group were obviously higher than those in the control. Subsequently, the flavonoids of ARs were extracted by flash extraction, where the optimized extraction process was: 63% ethanol, 69 s of extraction time, and a 57 mL/g liquid-material ratio. The findings provide a reference for the further industrial production of flavonoid-enriched O. elatus ARs, and the cultured ARs have potential application for the future production of products.

A New Species of Free-living Marine Nematode (Ptycholaimellus: Chromadoridae: Chromadorida: Nematoda) from Mangrove Wetlands in China.

This study presents a new species of free-living marine nematode, Ptycholaimellus luoyang sp. nov., from mangrove wetlands in China. The identification was confirmed by analyzing morphological characteristics and three genes: COI, 18S rDNA, and 28S rDNA. This species is distinguished from allied species by its short cephalic setae, cylindrical pharynx with anterior swelling, sclerotized transverse ridges occurring near the dorsal tooth, rod-like gubernaculum and proximal, arch-like, slightly waved, middle curved, and distally pointed spicules. The Bayesian topology was regarded as morphological evidence of P. luoyang sp. nov. being a distinct species. Interspecific and intrageneric thresholds of the K2P distance divergence have been presented here for the first time.

METTL3 modulates m6A modification of CDC25B and promotes head and neck squamous cell carcinoma malignant progression.

BACKGROUND: N6-methyladenosine (m6A) RNA methylation and its methyltransferase METTL3 have been widely reported to be involved in different cancers by regulating RNA metabolism and function. Here, we aimed to explore the biological function and clinical significance of m6A modification and METTL3 in head and neck squamous cell carcinoma (HNSCC). METHODS: The prognostic value of METTL3 expression was evaluated using tissue microarray and immunohistochemical staining analyses in a human HNSCC cohort. The biological role and mechanism of METTL3 in HNSCC tumour growth, metastasis and angiogenesis were determined in vitro and in vivo. RESULTS: M6A levels and METTL3 expressions in HNSCC tissues were significantly increased compared with paired adjacent tissues. Meanwhile, METTL3 was an independent risk factor for the prognosis of HNSCC patients. Moreover, METTL3 overexpression promoted HNSCC cell proliferation, migration, invasion, and angiogenesis, while knockdown of METTL3 had an opposite effect in vivo and in vitro. Mechanistically, METTL3 enhanced the m6A modification of CDC25B mRNA, which maintained its stability and upregulated its expression, thereby activating G2/M phase of cell cycle and leading to HNSCC malignant progression. CONCLUSIONS: METTL3 may be a potential prognostic biomarker and therapeutic target for HNSCC.

Resonant sheath heating in weakly magnetized capacitively coupled plasmas due to electron-cyclotron motion.

An electron heating mechanism based on a resonance between the cyclotron motion of electrons and the radio frequency sheath oscillations is reported in weakly magnetized capacitively coupled plasmas at low pressure. If half of the electron cyclotron period coincides with the radio frequency period, then electrons will coherently collide with the expanding sheath and gain substantial energy, which enhances the plasma density. A relation between the magnetic field and the driving frequency is found to characterize this resonance effect and the kinetics of electrons are revealed at resonance conditions for various driving frequencies.

Ultrasound-Guided Percutaneous Needle Biopsy for Peripheral Pulmonary Lesions: Diagnostic Accuracy and Influencing Factors.

A retrospective study was carried out to evaluate the diagnostic accuracy and the factors influencing the diagnostic accuracy of 648 procedures of ultrasound-guided percutaneous needle biopsy (PNB) for peripheral pulmonary lesions (PPLs). We reviewed the histopathology results, the clinical records and the procedure reports of these 648 biopsies and the final diagnoses of 637 PPLs to determine the diagnostic accuracy of ultrasound-guided PNB. Factors that influenced the diagnostic accuracy were assessed by analysis of the biopsy procedures, which were classified as diagnostic cases (true-positive and true-negative) and non-diagnostic cases (false-positive, false-negative and indeterminate). Statistical analyses of factors that related to patient demographic characteristics, lesion characteristics and biopsy details were performed to determine possible effects on diagnostic accuracy. Biopsies were successfully performed in all cases, and 11 patients underwent second biopsies for the same lesions. Among the 637 PPLs, there were 326 (51.2%) malignant lesions, 272 (42.7%) benign lesions and 39 (6.1%) indeterminate lesions. Of the 272 benign lesions, 114 (41.9%) were found to be tuberculous. The overall diagnostic accuracy was 81.8%, and the rates of hemoptysis, symptomatic pneumothorax and chest-tube insertion were 8.0%, 1.7% and 0.9%, respectively. Lesions sizes were divided into 3 groups according to the measurement by ultrasound. For lesions that measured ≤20 mm, 21-49 mm and ≥50 mm, the diagnostic accuracy was 72.0%, 86.8% and 79.7%, while sensitivity and specificity were 54.3%-79.2%, 88.3%-90.7% and 79.4%-89.5% and 77.3%-100%, 96.8%-100% and 58.6%-100%, respectively. Diagnostic accuracy was significantly affected by lesion size when lesion size was measured by ultrasound (p = 0.006) and computed tomography (CT) (p = 0.001). In the 3 lesion groups of ≤20 mm, 21-49 mm or ≥50 mm, diagnostic accuracy among each group was significantly different (p <0.001). When lesion size was measured by ultrasound (p <0.001) and CT (p <0.001) and the 3 groups were analyzed (p <0.001), there was a statistically significant relationship between lesion size and the presence of necrosis. The rates of the presence of necrosis in lesions that measured ≤20 mm, 21-49 mm and ≥50 mm were 3.9%, 11.7% and 28.8%, respectively. No significance was found for age (p = 0.119), gender (p = 0.25), lesion location (p = 0.55), the presence of necrosis (p = 0.226), patient position (p = 0.25), needle size (p = 0.26), puncture angle (p = 0.34) and needle passes (p = 0.21). Ultrasound-guided PNB is an effective and safe diagnostic method for PPLs; the diagnostic accuracy is significantly affected by lesion size and decreases in smaller (≤20 mm) and larger (≥50 mm) lesions.

Efficacy, safety and influencing factors of intra-calf muscular injection of bone marrow mononuclear cells in the treatment of type 2 diabetes mellitus-induced lower extremity vascular disease.

The efficacy, safety and impact of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) associated with the intra-calf muscular injection of bone marrow mononuclear cells (BMMCs) in the treatment of type 2 diabetes mellitus (T2DM)-induced lower extremity vascular disease (LEVD) were evaluated. Patients with T2DM-LEVD were randomly divided into a control group and BMMCs group to assess the efficacy and safety of the treatment; serum VEGF and bFGF levels were detected. The BMMCs group was divided into superior genicular artery (SGA) and inferior genicular artery (IGA) subgroups as well as low-dose and high-dose subgroups for the comparison of efficacy indices. The BMMCs group exhibited significantly improved indices (P<0.05) compared with the control group and no fatalities or cancer occurred. There were no significant changes in serum VEGF and bFGF levels (P>0.05). The claudication distance in the IGA subgroup was significantly greater that in the SGA subgroup (P<0.05); the low-dose subgroup and the high-dose subgroup did not demonstrate any significant differences in each index (P>0.05). BMMC treatment for T2DM-LEVD was found to be safe and effective and had no significant impact on serum VEGF and bFGF levels in the short term; However, the degree of LEVD may affect its efficacy.

Co-expression of Cyanobacterial Genes for Arsenic Methylation and Demethylation in Escherichia coli Offers Insights into Arsenic Resistance.

Arsenite [As(III)] and methylarsenite [MAs(III)] are the most toxic inorganic and methylated arsenicals, respectively. As(III) and MAs(III) can be interconverted in the unicellular cyanobacterium Nostoc sp. PCC 7120 (Nostoc), which has both the arsM gene (NsarsM), which is responsible for arsenic methylation, and the arsI gene (NsarsI), which is responsible for MAs(III) demethylation. It is not clear how the cells prevent a futile cycle of methylation and demethylation. To investigate the relationship between arsenic methylation and demethylation, we constructed strains of Escherichia coli AW3110 (ΔarsRBC) expressing NsarsM or/and NsarsI. Expression of NsarsI conferred MAs(III) resistance through MAs(III) demethylation. Compared to NsArsI, NsArsM conferred higher resistance to As(III) and lower resistance to MAs(III) by methylating both As(III) and MAs(III). The major species found in solution was dimethylarsenate [DMAs(V)]. Co-expression of NsarsM and NsarsI conferred As(III) resistance at levels similar to that with NsarsM alone, although the main species found in solution after As(III) biotransformation was methylarsenate [MAs(V)] rather than DMAs(V). Co-expression of NsarsM and NsarsI conferred a higher level of resistance to MAs(III) than found with expression of NsarsM alone but lower than expression of only NsarsI. Cells co-expressing both genes converted MAs(III) to a mixture of As(III) and DMAs(V). In Nostoc NsarsM is constitutively expressed, while NsarsI is inducible by either As(III) or MAs(III). Thus, our results suggest that at low concentrations of arsenic, NsArsM activity predominates, while NsArsI activity predominates at high concentrations. We propose that coexistence of arsM and arsI genes in Nostoc could be advantageous for several reasons. First, it confers a broader spectrum of resistance to both As(III) and MAs(III). Second, at low concentrations of arsenic, the MAs(III) produced by NsArsM will possibly have antibiotic-like properties and give the organism a competitive advantage. Finally, these results shed light on the role of cyanobacteria in the arsenic biogeochemical cycle.

Three new and two known free-living marine nematode species of the family Ironidae from the East China Sea.

Three new and two known species of free-living marine nematodes of the family Ironidae from the East China Sea are described and illustrated. Conilia sinensis sp. nov. is identified by the relatively large body size (1883-2399 µm); the well developed lips; the number, shape and length of spicule (single and striated, length 87-100 µm as arc); the shape of telamon; the number of supplements (1). Pheronous donghaiensis sp. nov. is characterized by its sharp tail point; caudal gland absent; buccal cavity armed with four big solid teeth and rows of minute denticles; spicules stout, with central septum at proximal end, male caudal region with two rows of small conical subventral papillae. Trissonchulus latispiculum sp. nov. can be distinguished by its head not set off from remaining body, tail short and blunt, buccal cavity with minute denticles, spinneret opening slightly ventrally, spicule broad and alate with central septum and head on proximal end. Trissonchulus benepapillosus (Schulz, 1935) and Trissonchulus oceanus Cobb 1920 which are first reported from China, are redescribed in detail with emphasis on new or hitherto poorly described morphological features. Types are deposited in the Institute of Oceanology, Chinese Academy of Sciences.

Centipeda minima (Ebushicao) extract inhibits PI3K-Akt-mTOR signaling in nasopharyngeal carcinoma CNE-1 cells.

BACKGROUND: Centipeda minima (Ebushicao) has been used for the treatment of various diseases, such as nasal allergies, rhinitis and sinusitis, nasopharyngeal carcinoma, cough, and headache. This study aims to investigate the anticancer activities of Centipeda minima ethanol extracts (CME) against nasopharyngeal carcinoma cell CNE-1 and their underlying mechanism. METHODS: CNE-1 cells were treated with different concentrations (15-50 µg/mL) of CME for different time intervals (24, 48, and 72 h). Cytotoxicity of CME was determined by MTT assay. Cell morphological changes were observed by fluorescence microscopy after HO 33258 staining. Cell cycle status was evaluated by flow cytometry following propidium iodide staining. Apoptosis was detected by flow cytometry following annexin V-FITC/PI staining. The levels of apoptosis-associated and PI3K-Akt-mTOR signaling related proteins were measured by western blotting analysis. RESULTS: CME (15-50 µg/mL) significantly inhibited the proliferation of CNE-1 in a dose- and time-dependent manner (P = 0.026 for 15 µg/mL, P < 0.001 for 25, 30, 40, and 50 µg/mL, respectively); the IC50 values (µg/mL) were 41.57 ± 0.17, 30.34 ± 0.06 and 24.98 ± 0.08 for 24, 48 and 72 h treatments, respectively. Significant morphological changes of CNE-1 cells displaying apoptosis were observed after CME treatment. CME showed low cytotoxicity toward normal LO2 cells. CNE-1 cells were arrested in the G2/M phase while treated with 15, 25, 40 µg/mL of CME, respectively (P = 0.032, P = 0.0053, P < 0.001). CME (15, 25, 40 µg/mL) down-regulated Bcl-2 expression (P = 0.032, P = 0.0074, P < 0.001), and up-regulated Bax (P = 0.026, P = 0.0056, P < 0.001) with activation of caspase-3, caspase-8, caspase-9, and PARP observed in CNE-1 cells (P = 0.015, P = 0.0067, P < 0.001 for caspase 3; P = 0.210, 0.028, < 0.001 for caspase 8; P = 0.152, 0.082, 0.0080 for caspase 9; P = 0.265, 0.0072, < 0.001 for PARP). CME suppressed the activation of the PI3K-AKT-mTOR pathway (P = 0.03, 0.0007, 0.004, 0.006, 0.022 for p-PI3K, p-Akt-Ser(473), p-Akt-Thr(308), p-mTOR-Ser(2448), p-mTOR-Ser(2481), respectively after 40 µg/mL of CME treated for 24 h). CONCLUSION: CME inhibited the proliferation of CNE-1 cells and activation of the PI3K-AKT-mTOR signaling pathway.

[Analysis of Applying Chinese Medical Clinical Pathway for Treating Attention-deficit Hyperactivity Disorder].

OBJECTIVE: To evaluate the application effect of Chinese medical clinical pathway for treating attention-deficit hyperactivity disorder (ADHD), and to provide evidence for further improving clinical pathways. METHODS: Totally 270 ADHD children patients were recruited and treated at pediatrics clinics of 9 cooperative hospitals from December 2011 to December 2012. The treatment course for all was 3 months. Scores of attention deficit and hyperactivity rating scale, scores of behavior, Conners index of hyperactivity (CIH), and Chinese medical syndrome scores were compared between before and after treatment. The efficacy difference in various sexes, ages, and disease courses were evaluated by judging standards for Chinese medical syndrome and ADHD. RESULTS: Fifteen children patients who entered clinical pathway dropped out, and the rest 255 completed this trial. Compared with before treatment, total scores of attention deficit and hyperactivity rating scale, scores of attention deficit and hyperactivity rating scale, CIH, and Chinese medical syndrome scores obviously decreased (all P < 0.01). The total effective rate in disease efficacy was 87.8% (224/255 cases), and the total effective rate in Chinese medical syndrome curative effect was 87.5% (223/255 cases). The clinical curative effect was not influenced by age, gender, or course of disease when statistically analyzed from judging standards for Chinese medical syndrome or for disease efficacy. CONCLUSION: Intervention by Chinese medical clinical pathway could improve ADHD patients' symptoms, and its efficacy was not influenced by sex, age, or course of disease.

Ethanolic extract of rhizome of Ligusticum chuanxiong Hort. (chuanxiong) enhances endothelium-dependent vascular reactivity in ovariectomized rats fed with high-fat diet.

The rhizome of Ligusticum chuanxiong Hort. (LC), also known as chuanxiong, is a very common herb widely used to treat cardiovascular and cerebrovascular diseases. It is also used as a major ingredient in soups for regular consumption to promote good health. To study the protective effect of LC ethanolic extract (LCEE, 600 mg per kg per day, p.o.) on the integrity of the vascular system, ovariectomized (OVX) rats were fed with a high-fat diet (HFD) plus LCEE for 12 weeks. The animal model was used to mimic the dyslipidemic condition seen in postmenopausal women. LCEE was found significantly to reduce the body weight gain, improve serum lipid profiles (by lowering total cholesterol and low density lipoprotein cholesterol but raising high density lipoprotein cholesterol) and protect vascular endothelium in the HFD-fed OVX rats. It is postulated that LCEE could exert its vascular protective effect through multiple targets by (1) improving serum lipid profiles to reduce the detrimental effects of cholesterol; (2) reducing the ROS level in the body via enhancing the hepatic anti-oxidative activity or antioxidant level to scavenge the reactive oxygen species generated in the postmenopausal hypercholesterolemic condition; (3) stimulating eNOS-derived nitric oxide production; (4) counteracting the up-regulation of inflammatory cytokine (TNF-α, VCAM-1 and ICAM-1) expressions so as to reduce endothelium damage.

[Distribution of compact bone mesenchymal stem cells in lung tissue and bone marrow of mouse].

This study was aimed to investigate the distribution of compact bone mesenchymal stem cells(MSC) marked with lentiviral plasmid pGC FU-RFP-LV in lung tissue and bone marrow of mouse. The MSC were infected by lentivirus with infection efficiency 78%, the infected MSC were injected into BALB/c mice via tail veins in concentration of 1×10(6) /mouse. The mice were randomly divided into 4 group according to 4 time points as 1, 2, 5 and 7 days. The lung tissue and bone marrow were taken and made of frozen sections and smears respectively in order to observed the distributions of MSC. The results indicated that the lentiviral infected MSC displayed phenotypes and biological characteristics which conformed to MSC by immunophenotyping analysis and induction differentiation detection. After the MSC were infected with optimal viral titer MOI = 50, the cell growth no significantly changed; the fluorescent microscopy revealed that the distributions of MSC in bone marrow on day 1, 2, 5 and 7 were 0.50 ± 0.20, 0.67 ± 0.23, 0.53 ± 0.14, 0.33 ± 0.16; those in lung tissue were 0.55 ± 0.15, 0.47 ± 0.13, 0.29 ± 0.13, 0.26 ± 0.08. It is concluded that the distribution of MSC in lung tissue reaches a peak on day 1, while distribution of MSC in bone marrow reaches a peak on day 2. The distribution of mouse MSC relates with RFP gene expression and implantation of MSC in lung tissue and bone marrow.

[Research progress on immunologic mechanisms of mesenchymal stem cells for treatment of graft-versus-host disease].

Mesenchymal stem cells (MSC) are the non-hematopoietic stem cells with a multi-differentiation potentials, which has a low immunogenicity and immune regulation ability. MSC immune regulation ability is particularly important, such as MSC can inhibit the activation and proliferation of T, B lymphocytes, NK cells and dendritic cells (DC). Meanwhile, MSC is able to reconstruct the human hematopoietic microenvironment, improving the successful rate of hematopoietic stem cell transplantation. Graft versus host disease (GVHD) is the main factor causing hematopoietic stem cell transplantation-related mortality. Based on the above mentioned properties, MSCs are used to treat autoimmune diseases and GVHD, recently. Therefore, deep exploration of the cellular immune mechanisms of MSC to treat GVHD is particularly important. This review focuses on progress of research related to treatment of GVHD by MSC immune mechanisms and briefly summarizes the status of the clinical studies.

Freeliving marine nematodes as a pollution indicator of the Bohai Sea.

A hierarchical diversity index--taxonomic distinctness index delta+, which was first defined by Warwick and Clark in 1998, was employed to evaluate the pollution status of the Bohai Sea with freeliving marine nematodes. The result showed that the Bohai Bay and other coastal sampling sites might be affected by oil and gas production and other anthropogenic influences. In other words, anthropogenic disturbance was affecting this component of the benthos in these locations. And most offshore sampling sites in the middle of the Bohai Sea were clear and unpolluted.