Transcriptome and Metabolome Integration Reveals the Impact of Fungal Elicitors on Triterpene Accumulation in Sanghuangporus sanghuang.

Sanghuangporus sanghuang is a large wood-decaying mushroom highly valued in traditional Chinese medicine due to its medicinal properties, including hypoglycemic, antioxidant, antitumor, and antibacterial properties effects. Its key bioactive compounds include flavonoids and triterpenoids. Specific fungal genes can be selectively induced by fungal elicitors. To investigate the effect of fungal polysaccharides derived from Perenniporia tenuis mycelia on the metabolites of S. sanghuang, we conducted metabolic and transcriptional profiling with and without elicitor treatment (ET and WET, respectively). Correlation analysis showed significant differences in triterpenoid biosynthesis between the ET and WET groups. In addition, the structural genes associated with triterpenoids and their metabolites in both groups were verified using quantitative real-time polymerase chain reaction (qRT-PCR) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Through metabolite screening, three triterpenoids were identified: betulinol, betulinic acid, and 2-hydroxyoleanolic acid. Excitation treatment increased the level of betulinic acid by 2.62-fold and 2-hydroxyoleanolic acid by 114.67-fold compared to WET. The qRT-PCR results of the four genes expressed in secondary metabolic pathways, defense gene activation, and signal transduction showed significant variation between the ET and WET groups. Overall, our study suggests that the fungal elicitor induced the aggregation of pentacyclic triterpenoid secondary metabolites in S. sanghuang.

Long-term phosphorus addition alters plant community composition but not ecosystem stability of a nitrogen-enriched desert steppe.

Under ongoing global change, whether grassland ecosystems can maintain their functions and services depends largely on their stability. However, how ecosystem stability responds to increasing phosphorus (P) inputs under nitrogen (N) loading remains unclear. We conducted a 7-year field experiment to examine the influence of elevated P inputs (ranging from 0 to 16 g P m-2 yr-1) on the temporal stability of aboveground net primary productivity (ANPP) under N addition of 5 g N·m-2·yr-1 in a desert steppe. We found that under N loading, P addition altered plant community composition but did not significantly affect ecosystem stability. Specifically, with the increase in the P addition rate, declines in the relative ANPP of legume could be compensated for by an increase in the relative ANPP of grass and forb species, yet community ANPP and diversity remained unchanged. Notably, the stability and asynchrony of dominant species tended to decrease with increasing P addition, and a significant decrease in legume stability was observed at high P rates (>8 g P m-2 yr-1). Moreover, P addition indirectly affected ecosystem stability by multiple pathways (e.g., species diversity, species asynchrony, dominant species asynchrony, and dominant species stability), as revealed by structural equation modeling results. Our results suggest that multiple mechanisms work concurrently in stabilizing the ecosystem stability of desert steppes and that increasing P inputs may not alter desert steppe ecosystem stability under future N-enriched scenarios. Our results will help improve the accuracy of vegetation dynamics assessments in arid ecosystems under future global change.

Characterizing Diversity Based on the Chemical and Nutritional Composition of Shiitake Culinary-Medicinal Mushroom Lentinula edodes (Agaricomycetes) Commonly Cultivated in China.

A comparative study was carried out on the chemical composition and nutritional value of six cultivated Lentinula edodes strains that are widely appreciated in China. The results demonstrated that all investigated L. edodes were good sources of protein (14.87%-27.13%), carbohydrates (62.03%-75.56%), and dietary fiber (35.88%-42.49%) and had low ash (5.24%-6.38%) and low fat (0.80%-1.70%) content. There were significant differences among different cultivars. Shenxiang 215 had high crude protein and dietary fiber contents. Potassium was the most abundant mineral element, followed by phosphorus. Different cultivars exhibited distinct fatty acid compositions and free amino acid profiles. Shenxiang 215 had high essential amino acid content. Polyunsaturated fatty acids were predominant in cultivars 0912, Huxiang F4, and Huxiang F2; however, monounsaturated fatty acids were predominant in other strains. Cultivar 0912 had a better mineral and polyunsaturated fatty acid profile. The amino acid profile and protein quality were systematically investigated referring to the latest version of international amino acid reference patterns, including the amino acid score, ratio coefficient of amino acid, ratio coefficient score of amino acid, essential amino acid index, and protein digestibility - corrected amino acid score. The results demonstrated that Shenxiang 18 had better protein quality. These findings provide a reference for breeders to select parents for directional quality breeding.

Effect of combination therapy with neural stem cell transplantation and teramethylpyrazine in rats following acute spinal cord injury.

OBJECTIVES: This study was to explore the effects of teramethylpyrazine (TMP) administered in conjunction with neural stem cell transplantation on motor function, pathological lesions and the Janus kinase (JAK)2/signal transducer and activator of transcription 3 signal transduction pathway in rats following acute spinal cord injury (SCI). METHODS: Female Sprague-Dawley rats were randomly divided into sham, model, neural stem cells (NSCs) and NSCs+TMP groups. Motor function was evaluated using the Basso, Beattie, Bresnahan scale. Spinal cord neuropathies and neuron apoptosis were observed by HE and TUNEL staining. The brain-derived neurotrophic factor (BDNF), Nogo-A, JAK2 and p-JAK2 protein levels were measured by western blot analysis. RESULTS: NSCs+TMP significantly improved rat motor function, attenuated impaired spinal cords, and decreased cellular apoptosis, compared with NSCs therapy alone (P < 0.05). In addition, expression of BDNF protein was significantly higher in NSCs+TMP rats compared with other groups regardless of time postinjury (P < 0.05). The highest expression levels of Nogo-A protein were observed in the model group. The expression of p-JAK2 in the NSCs+TMP group was relatively lower than the model and NSCs groups (P < 0.05). CONCLUSIONS: In rats with SCI, NSCs+TMP effectively improved motor function and offered spinal cord protection by increasing BDNF and decreasing Nogo-A levels, as well as inhibiting the JAK2/STAT3 signal transduction pathway, suggesting that TMP could be a useful agent in NSCs transplantation in the treatment of SCI.

Overexpression of the saccharopine dehydrogenase gene improves lysine biosynthesis in Flammulina velutipes.

Saccharopine dehydrogenase (EC 1.5.1.7) regulates the last step of fungal lysine biosynthesis. The gene (Fvsdh) encoding saccharopine dehydrogenase was identified and cloned from the whole genome of Flammulina velutipes. The genomic DNA of Fvsdh is 1257 bp, comprising three introns and four exons. The full-length complementary DNA of Fvsdh comprises 1107 bp with a deduced amino acid sequence of 368 residues. A 1,000-bp promoter sequence containing the TATA box, CAAT box, and several putative cis-acting elements was also identified. The results of tissue expression analysis showed that the expression level of the Fvsdh gene was higher in the pileus than in the stipe whether in the elongation or maturation stage. Further research showed that the lysine contents were 3.03 and 2.95 mg/g in maturation-pileus and elongation-pileus, respectively. In contrast, the lysine contents were 2.49 and 2.07 mg/g in elongation-stipe and maturation-stipe, respectively. To study the function of Fvsdh, we overexpressed Fvsdh in F. velutipes and found that Fvsdh gene expression was increased from 1.1- to 3-fold in randomly selected transgenic strains. The lysine contents were also increased from 1.12- to 1.3-fold in these five transformants, except for strain T3, in which the lysine contents were the same as the control. These results indicate that the expression of the Fvsdh gene can affect the lysine content of F. velutipes.

Preparation of molecularly imprinted polymers coupled with high-performance liquid chromatography for the selective extraction of salidroside from Rhodiola crenulata.

Salidroside is one of the bio-active compounds found in Rhodiola crenulata. To find an easy, time saving and efficient way to extract, purify and enrich salidroside from Rhodiola and other natural plants, we prepared a highly selective molecularly imprinted polymer (MIP) for extraction and preconcentration of salidroside using salidroside (SD) as a template, acrylamide (AM) as a functional monomer, ethylene glycol dimethacrylate (EDMA) as a crosslinking monomer, and dimethyl formamide (DMF) as a porogen. The performance of the MIPs was evaluated through selective recognition capacity and adsorption isotherms and kinetics. The results showed that MIPs possessed excellent specific recognition toward SD and could effectively discriminate its structural analogue. The application of the developed MIPs as a selective sorbent for solid-phase extraction (SPE) of SD was also investigated. Under the optimum conditions, a rapid, economical, and efficient method based upon MIP-SPE coupled with high-performance liquid chromatography (HPLC) was developed for the determination of SD in Rhodiola crenulata. The method showed satisfactory recoveries (from spiked real samples at 3 fortification levels of 0.5, 1 and 10 mg L-1) of 88.74%- 97.64% with relative standard deviations (RSDs) ranging from 2.05%-3.54%. Furthermore, MIP-SPE was successfully used to separate and purify SD from different parts in Rhodiola crenulata and it should be available for determination of salidroside in others herbs.

Changes in C:N:P stoichiometry modify N and P conservation strategies of a desert steppe species Glycyrrhiza uralensis.

Numerous studies have concluded that carbon (C):nitrogen (N):phosphorus (P) stoichiometry in both soils and plants tends to be decoupled under global change. We consequently hypothesized that plants will adjust nutrient conservation strategies to balance the altered elemental stoichiometry accordingly. To test our hypothesis, we conducted two pot-cultured experiments (with 8-level water and 6-level N addition treatments) using N-fixing species Glycyrrhiza uralensis Fisch from a desert steppe in northwestern China. We observed that high water availability lowered total N content and the N:P ratio in soils, further promoting both N and P resorption from senescing leaves of G. uralensis. High N addition enhanced soil N availability and the N:P ratio, thereby reducing N resorption, but increasing P resorption of G. uralensis. Comparatively, there were also great changes in senescing leaf C:N:P stoichiometry while no clear changes were observed in either green leaf or root C:N:P stoichiometry of G. uralensis. As expected, the altered C:N:P stoichiometry may, in turn, modify N and P conservation strategies through their close linkages with N and P uptake in green leaves of G. uralensis. This modification may also further exert effects on N and P cycling of the desert steppe.

Ophiopogonin D improves osteointegration of titanium alloy implants under diabetic conditions by inhibition of ROS overproduction via Wnt/ß-catenin signaling pathway.

A high failure rate of titanium implants in diabetic patients has been indicated in clinical evidences. Excessive oxidative stress at the bone-implant interface plays an important role in the impaired osteointegration under diabetic conditions. While the underlying mechanisms remain unknown and the targeted treatments are urgently needed. Ophiopogonin D (OP-D), isolated from Chinese herbal Radix Ophiopogon japonicus, is generally reported to be a potent antioxidant agent. In the present study, we hypothesized that OP-D exerted promotive effects on osteointegration against oxidative stress, and investigated the underlying mechanisms associated with alteration of Wnt/ß-catenin signaling pathway. Rabbit osteoblasts incubated on titanium alloy implant were co-cultured with normal serum (NS), diabetic serum (DS), DS + OP-D, DS + NAC (a potent ROS inhibitor) and DS + OP-D + Dkk1 (a Wnt inhibitor) for examinations of osteoblast behaviors. For in vivo study, titanium alloy implants were implanted into the femoral condyle defects on diabetic rabbits. Results demonstrated that diabetes-induced oxidative stress resulted in osteoblast dysfunctions and apoptotic injury at the bone-implant interface, concomitant with the inactivation of Wnt/ß-catenin signaling. Importantly, OP-D administration attenuated oxidative stress, directly reactivating Wnt/ß-catenin signaling. Osteoblast dysfunctions were thus reversed as evidenced by improved osteoblast adhesion, proliferation and differentiation, and ameliorated apoptotic injury, exerting similar effects to NAC treatment. In addition, the positive effects afforded by OP-D were confirmed by improved osteointegration and oetogenesis within the titanium alloy implants in vivo by Micro-CT and histological analyses. Furthermore, the pro-osteogenic effects of OP-D were almost completely abolished by the Wnt inhibitor Dkk1. These results demonstrated, for the first time, OP-D administration alleviated the damaged osteointegration of titanium alloy implants under diabetic conditions by means of inhibiting oxidative stress via a Wnt/ß-catenin-dependent mechanism. The OP-D administration would become a reliable treatment strategy for implant failure therapy in diabetics due to the optimal anti-oxidative and pro-osteogenic properties.

Phosphorus addition changes belowground biomass and C:N:P stoichiometry of two desert steppe plants under simulated N deposition.

Many studies have reported that increasing atmospheric nitrogen (N) deposition broadens N:phosphorus (P) in both soils and plant leaves and potentially intensifies P limitation for plants. However, few studies have tested whether P addition alleviates N-induced P limitation for plant belowground growth. It is also less known how changed N:P in soils and leaves affect plant belowground stoichiometry, which is significant for maintaining key belowground ecological processes. We conducted a multi-level N:P supply experiment (varied P levels combined with constant N amount) for Glycyrrhiza uralensis (a N fixing species) and Pennisetum centrasiaticum (a grass) from a desert steppe in Northwest China during 2011-2013. Results showed that increasing P addition increased the belowground biomass and P concentrations of both species, resulting in the decreases in belowground carbon (C):P and N:P. These results indicate that P inputs alleviated N-induced P limitation and hence stimulated belowground growth. Belowground C:N:P stoichiometry of both species, especially P. centrasiaticum, tightly linked to soil and green leaf C:N:P stoichiometry. Thus, the decoupling of C:N:P ratios in both soils and leaves under a changing climate could directly alter plant belowground stoichiometry, which will in turn have important feedbacks to primary productivity and C sequestration.

[Analysis on adverse event in acupotomy therapy based on literature research].

OBJECTIVE: To explore the current domestic situation of the adverse event in acupotomy so as to promote the standardization of acupotomy and improve the clinical effect and safety of acupotomy therapy. METHODS: The method of retrospective journal researching was used. The case report about the adverse event in acupotomy from CNKI, WANFANG and VIP databases was retrieved and the types, the features, the causes, the numbers of the papers and the cases were analyzed. RESULTS: Fifty-eight papers were included, with total 343 cases. It was indicated that the adverse events in acupotomy were mainly postoperative local pain, hematoma, swelling, nerve damage, tendon rupture, adhesion, injury, infection, syncope, anesthesia accident, undeserved hormone use, etc. The lower level of standardization of acupotomy in the clinical use may lead to adverse events. CONCLUSION: Safe acupotomy treatment for some complicated diseases and dangerous parts remains to be improved, and the standardization of the acupotomy is an important approach to improve its safety. The related work needs to be carried out.

A summary and evaluation of current evidence for myocardial infarction with Chinese medicine.

OBJECTIVE: To provide evidence-based recommendations for clinical application and provoke thoughts for future researchers by conducting a comprehensive summary and evaluation of the current evidence profile for the role of Chinese medicine (CM) in treating myocardial infarction (MI). METHODS: Online databases including PubMed, EMBase, Cochrane Library, Chinese National Knowledge Infrastructure (CNKI), Chinese Biomedical Medicine (CBM), VIP Journal Integration Platform, and Wanfang database were systematically searched for literatures on CM in treating MI. After screening, studies were categorized into 5 types, i.e. systematic review (SR), randomized controlled trial (RCT), observational study, case report and basic research. General information was abstracted, and the quality levels of these studies and their conclusions were summarized and assessed. RESULTS: A total of 452 studies including 10 SRs, 123 RCTs, 47 observational studies, 28 case reports, and 244 basic researches were selected. Clinical studies centered primarily on herbal decoction and mostly were not rigorously performed. High-quality studies were predominantly on Chinese patent medicines (CPMs) such as Danshen Injection (), Shenmai Injection (), Shengmai Injection () and Qishen Yiqi Dripping Pills (). The most frequently observed pattern of drug combination was decoction plus injection. Results of SRs and clinical studies showed that CM may reduce mortality, decrease risk of complication, reduce myocardial injury, improve cardiac function and inhibit ventricular remodeling. Findings from basic researches also supported the positive role of CM in reducing infarct size and myocardial injury, promoting angiogenesis, preventing ventricular remodeling and improving cardiac function. According to the current evidence body, CM has proven effects in the prevention and treatment of MI. It is also found that the effects of CPMs vary with indications. For instance, Shenmai Injection has been found to be especially effective for reducing the incidence of acute clinical events, while CPMs with qi-nourishing and bloodcirculating properties have been proven to be effective in inhibiting ventricular remodeling. High quality evidence supports the use of CM injection for acute MI and CPM for secondary prevention. Reports on adverse events and other safety outcomes associated with CM for MI are scarce. CONCLUSIONS: Sufficient evidence supported the use of CM as an adjuvant to Western medicine for preventing and treating MI. The choice of drug use varies with disease stage and treatment objective. However, the quality of the evidence body remains to be enhanced.

Oleuropein, a natural extract from plants, offers neuroprotection in focal cerebral ischemia/reperfusion injury in mice.

Oleuropein (OLE) was found to have anti-inflammatory and anti-oxidant effects. The latest study has shown that it can resist myocardial injury that follows an acute myocardial infarction and can rescue impaired spinal nerve cells. In this study, we investigated the neuroprotective effects of OLE on cerebral ischemia and reperfusion injury in a middle cerebral artery occlusion model in mice.OLE (100 mg/kg) was injected intraperitoneally 1h before ischemia. We found that the volume of cerebral infarction was significantly reduced after 75 min of ischemia and 24 h of reperfusion compared with the I/R (ischemia/reperfusion) group. This protective function occurred in a dose-dependent manner. We also found that treatment with OLE could reduce the cerebral infarct volume. The neuroprotective effect was prolonged from 2 h to 4 h when we injected OLE intracerebroventricularly after reperfusion. We then found that OLE can decrease the level of cleavedcaspase-3, an important marker of apoptosis, in the ischemic mouse brain. Finally, we explored the role of OLE in providing anti-apoptotic effects through the increased expression of Bcl-2 and the decreased expression of Bax, which are important markers in apoptosis. As shown above, the function and safety of OLE in cardiovascular disease may indicate that it is a potential therapeutic for stroke.

Curcumin promotes nerve regeneration and functional recovery after sciatic nerve crush injury in diabetic rats.

Curcumin is capable of promoting peripheral nerve regeneration in normal condition. However, it is unclear whether its beneficial effect on nerve regeneration still exists under diabetic mellitus. The present study was designed to investigate such a possibility. Diabetes in rats was developed by a single dose of streptozotocin at 50 mg/kg. Immediately after nerve crush injury, the diabetic rats were intraperitoneally administrated daily for 4 weeks with curcumin (50 mg/kg, 100 mg/kg and 300 mg/kg), or normal saline, respectively. The axonal regeneration was investigated by morphometric analysis and retrograde labeling. The functional recovery was evaluated by electrophysiological studies and behavioral analysis. Axonal regeneration and functional recovery was significantly enhanced by curcumin, which were significantly better than those in vehicle saline group. In addition, high doses of curcumin (100 mg/kg and 300 mg/kg) achieved better axonal regeneration and functional recovery than low dose (50 mg/kg). In conclusion, curcumin is capable of promoting nerve regeneration after sciatic nerve crush injury in diabetes mellitus, highlighting its therapeutic values as a neuroprotective agent for peripheral nerve injury repair in diabetes mellitus.

Quality assessment of randomized controlled trials reporting on knee osteoarthritis treated with warming needle moxibustion.

OBJECTIVE: To evaluate the quality of randomized controlled trials (RCTs) testing the efficacy of warming needle moxibustion on knee osteoarthritis (KOA). METHODS: Nine databases were searched systematically including: the Cochrane Library (1/1993-10/2012), PubMed (1/1980-10/2012), EMBASE (1/1990-10/12012), SCI Expanded (1/1986-10/2012), Science Direct (1/1979-10/2012), Chinese Biomedical Literature Database disc (1/1978-10/2012), China National Knowledge Infrastructure Database (1/1979-10/2012), China Science and Technology Journal Database (a full text issue database of China, 1/1989-10/2012), and Wanfang Database (a full text issue database of China, 1/1990-10/12012). Traditional reference retrieval was also conducted. Language was limited to Chinese and English. We identified 37 RCTs that used warming needle moxibustion as an intervention and they were assessed with the Consolidated Standards for Reporting of Trials Statement 2010 (CONSORT 2010) and Standards for Reporting Interventions Controlled Trials of Acupuncture 2010 (STRICT A 2010). RESULTS: Of the 37 reports, only seventeen (45.95%) mentioned the random allocation of sequence methods, nine used unacceptable methods, and only one (2.70%) gave the description of the mechanism of allocation concealment. One study (2.70%) mentioned blinding, one (2.70%) had a sample size calculation, and twenty-three (62.16%) used reasonable statistical methods. Thirteen (35.14%) described the background and reason for the RCTs, three (8.11%) gave a description of lost or eliminated patients, and two (5.41%) reported accidental situations. Only three (8.11%) gave analysis of the RCT limitations. No report mentioned intentional analysis. CONCLUSION: The quality of the RCTs assessed in this study was from moderate to low. The design of RCTs, the methods of statistical analysis, and the description of reports information needs to be improved. CONSORT 2010 and STRICT A 2010 should be used to standardize the reporting of acupuncture RCTs in the future.

[Acupoint: the sensor of the information in the organic body].

Based on the structure and function of acupoint and in association of the definition and principle of sensor, the acupoint is the sensitive element, being sensitive to the physical stimulation with acupuncture and moxibustion and sensitively responded to the disorders; the acupoint is the sensing element, transforming the changes of the acupoint information via the complicated internet conduction, integration and regulation, so as to generate the effects on organic body; the acupoint is the conversion element, transforming every irritation into the bioelectric signal or optical signal so that the organic body could recognize it. Therefore, the acupoint is regarded as the sensor of information in the organic body.

Bioavailability and delivery of nutraceuticals using nanotechnology.

Nanotechnology is an enable technology that has the potential to revolutionize agriculture and food systems. Driven by increasing consumer demand for healthy food products, researchers have been applying tools and knowledge in nanotechnology to address the issues relevant to food and nutrition. This concise review is mainly focused on nanoemulsions and polymer micelles-based delivery systems which have shown enhanced oral bioavailability and biological efficacies (that is, antiinflammation, anti-cancer, and so on) of different phytochemicals. Nanoemulsions are a class of extremely small droplets that appear to be transparent or translucent with a bluish coloration. They are usually in the range 50 to 200 nm but much smaller than the range (from 1 to 100 mum) for conventional emulsions. Nanoemulsion preparation, characterization, and bioavailability have been discussed. Curcumin nanoemulsions show 85% inhibition of TPA-induced mouse ear inflammation as well as the inhibition of cyclin D1 expression, while dibenzoylmethane (DBM) nanoemulsion shows about 3-fold increase in oral bioavailability compared to the conventional DBM emulsion. Biopolymer micelles show significantly improved water solubility/dispersibility and in vitro anti-cancer activity of phytochemicals. More research efforts are still needed for the understanding of the potential impacts of nanoencapsulated phytochemicals on the human body and environment to address the public concerns.

Water supply changes N and P conservation in a perennial grass Leymus chinensis.

Changes in precipitation can influence soil water and nutrient availability, and thus affect plant nutrient conservation strategies. Better understanding of how nutrient conservation changes with variations in water availability is crucial for predicting the potential influence of global climate change on plant nutrient-use strategy. Here, green-leaf nitrogen (N) and phosphorus (P) concentrations, N- and P-resorption proficiency (the terminal N and P concentration in senescent leaves, NRP and PRP, respectively), and N- and P-resorption efficiency (the proportional N and P withdrawn from senescent leaves prior to abscission, NRE and PRE, respectively) of Leymus chinensis (Trin.) Tzvel., a typical perennial grass species in northern China, were examined along a water supply gradient to explore how plant nutrient conservation responds to water change. Increasing water supply at low levels (< 9000 mL/year) increased NRP, PRP and PRE, but decreased green-leaf N concentration. It did not significantly affect green-leaf P concentration or NRE. By contrast, all N and P conservation indicators were not significantly influenced at high water supply levels (> 9000 mL/year). These results indicated that changes in water availability at low levels could affect leaf-level nutrient characteristics, especially for the species in semiarid ecosystems. Therefore, global changes in precipitation may pose effects on plant nutrient economy, and thus on nutrient cycling in the plant-soil systems.