Hen Egg White Lysozyme (HEWL) Confers Resistance to Verticillium Wilt in Cotton by Inhibiting the Spread of Fungus and Generating ROS Burst.

Verticillium wilt is a soil-borne vascular disease caused by the fungal pathogen Verticillium dahliae. It causes great harm to upland cotton (Gossypium hirsutum) yield and quality. A previous study has shown that Hen egg white lysozyme (HEWL) exerts strong inhibitory activity against V. dahliae in vitro. In the current study, we introduced the HEWL gene into cotton through the Agrobacterium-mediated transformation, and the exogenous HEWL protein was successfully expressed in cotton. Our study revealed that HEWL was able to significantly inhibit the proliferation of V. dahlia in cotton. Consequently, the overexpression of HEWL effectively improved the resistance to Verticillium wilt in transgenic cotton. In addition, ROS accumulation and NO content increased rapidly after the V. dahliae inoculation of plant leaves overexpressing HEWL. In addition, the expression of the PR genes was significantly up-regulated. Taken together, our results suggest that HEWL significantly improves resistance to Verticillium wilt by inhibiting the growth of pathogenic fungus, triggering ROS burst, and activating PR genes expression in cotton.

Co-expression of nitrogenase proteins in cotton (Gossypium hirsutum L.).

Chemical nitrogen fertilizer can maintain crop productivity, but overuse of chemical nitrogen fertilizers leads to economic costs and environmental pollution. One approach to reduce use of nitrogen fertilizers is to transfer nitrogenase biosynthetic pathway to non-legume plants. Fe protein encoded by nifH and MoFe protein encoded by nifD and nifK are two structural components of nitrogenase. NifB encoded by nifB is a critical maturase that catalyzes the first committed step in the biosynthesis of nitrogenase FeMo-cofactor that binds and reduces N2. Expression of the nifB, nifH, nifD and nifK is essential to generate plants that are able to fix atmospheric N2. In this study, the four genes (nifB, nifH, nifD and nifK) from Paenibacillu polymyxaWLY78 were assembled in plant expression vector pCAMBIA1301 via Cre/LoxP recombination system, yielding the recombinant expression vector pCAMBIA1301-nifBHDK. Then, the four nif genes carried in the expression vector were co-introduced into upland cotton R15 using Agrobacterium tumefaciens-mediated transformation. Homozygous transgenic cotton lines B2, B5 and B17 of T3 generation were selected by PCR and RT-PCR. qRT-PCR showed that nifB, nifH, nifD and nifK were co-expressed in the transgenic cottons at similar levels. Western blotting analysis demonstrated that NifB, NifH, NifD and NifK were co-produced in the transgenic cottons. Co-expression of the four critical Nif proteins (NifB, NifH, NifD and NifK) in cottons represents an important step in engineering nitrogenase biosynthetic pathway to non-legume plants.

Metabolic Engineering of Model Microorganisms for the Production of Xanthophyll.

Xanthophyll is an oxidated version of carotenoid. It presents significant value to the pharmaceutical, food, and cosmetic industries due to its specific antioxidant activity and variety of colors. Chemical processing and conventional extraction from natural organisms are still the main sources of xanthophyll. However, the current industrial production model can no longer meet the demand for human health care, reducing petrochemical energy consumption and green sustainable development. With the swift development of genetic metabolic engineering, xanthophyll synthesis by the metabolic engineering of model microorganisms shows great application potential. At present, compared to carotenes such as lycopene and ß-carotene, xanthophyll has a relatively low production in engineering microorganisms due to its stronger inherent antioxidation, relatively high polarity, and longer metabolic pathway. This review comprehensively summarized the progress in xanthophyll synthesis by the metabolic engineering of model microorganisms, described strategies to improve xanthophyll production in detail, and proposed the current challenges and future efforts needed to build commercialized xanthophyll-producing microorganisms.

Comparison of Activity and Safety of DSPAα1 and Its N-Glycosylation Mutants.

DSPAα1 is a potent rude thrombolytic protein with high medicative value. DSPAα1 has two natural N-glycan sites (N153Q-S154-S155, N398Q-K399-T400) that may lead to immune responses when administered in vivo. We aimed to study the effect of its N-glycosylation sites on DSPAα1 in vitro and in vivo by mutating these N-glycosylation sites. In this experiment, four single mutants and one double mutant were predicted and expressed in Pichia pastoris. When the N398Q-K399-T400 site was mutated, the fibrinolytic activity of the mutant was reduced by 75%. When the N153Q-S154-S155 sites were inactivated as described above, the plasminogen activating activity of its mutant was reduced by 40%, and fibrin selectivity was significantly reduced by 21-fold. The introduction of N-glycosylation on N184-G185-A186T and K368N-S369-S370 also considerably reduced the activity and fibrin selectivity of DSPAα1. The pH tolerance and thermotolerance of all mutants did not change significantly. In vivo experiments also confirmed that N-glycosylation mutations can reduce the safety of DSPAα1, lead to prolonged bleeding time, non-physiological reduction of coagulation factor (α2-AP, PAI) concentration, and increase the risk of irregular bleeding. This study ultimately demonstrated the effect of N-glycosylation mutations on the activity and safety of DSPAα1.

New insights explain that organic agriculture as sustainable agriculture enhances the sustainable development of medicinal plants.

As global health care demand continues to increase, medicinal plant productivity must progress without exhausting critical environmental resources. Hence, it is important to explore practices that can improve the quality, safety, and sustainability of medicinal plants, as well as ecological stability. Organic farming has recently gained significance as a sustainable cultivation alternative owing to increased awareness of the adverse effects of conventional cultivation method. Here, this study aimed to investigate the feasibility of organic farming as a solution for sustainable cultivation of medicinal plants from multiple perspectives and long-term benefits to the environment. Organic agricultural practices of medicinal plants were evaluated from a multi-dimensional perspective (environment, economy, and society) using extensive research data and literature and field surveys. Data from medicinal plant cultivation in Inner Mongolia were acquired for 76 sites from four data stations between 2014 and 2021. Data analysis revealed that organic medicinal plants can improve safety by reducing pesticide exposure risks. Simultaneously, organic agriculture of medicinal plants can improve biodiversity by effectively reducing pesticide and fertilizer use, which also provides natural safe products for health care. With the improvement of quality, the retail price will have a certain advantage, which will improve the income of farmers. Moreover, organic agriculture enhanced profitability because of the higher organic premium on medicinal plant products and improved ecosystem stability by increasing plant diversity. The findings of this study suggest that organic cultivation strategies can improve the quality and safety of medicinal plants and further provide a basis for promoting the sustainable development and ecological stability of medicinal plants. However, not all medicinal plant cultivators are guaranteed to adopt organic farming practices, but if all technological elements are correctly applied, the system can be maintained sustainably to expand the area of organically cultivated plants in the future.

Different N-Glycosylation Sites Reduce the Activity of Recombinant DSPAα2.

Bat plasminogen activators α2 (DSPAα2) has extremely high medicinal value as a powerful natural thrombolytic protein. However, wild-type DSPAα2 has two N-glycosylation sites (N185 and N398) and its non-human classes of high-mannose-type N-glycans may cause immune responses in vivo. By mutating the N-glycosylation sites, we aimed to study the effect of its N-glycan chain on plasminogen activation, fibrin sensitivity, and to observe the physicochemical properties of DSPAα2. A logical structure design was performed in this study. Four single mutants and one double mutant were constructed and expressed in Pichia pastoris. When the N398 site was eliminated, the plasminogen activator in the mutants had their activities reduced to ~40%. When the N185 site was inactivated, there was a weak decrease in the plasminogen activation of its mutant, while the fibrin sensitivity significantly decreased by ~10-fold. Neither N-glycosylation nor deglycosylation mutations changed the pH resistance or heat resistance of DSPAα2. This study confirms that N-glycosylation affects the biochemical function of DSPAα2, which provides a reference for subsequent applications of DSPAα2.

Xiaoke Decoction in treatment of type II diabetes: A Meta-analysis.

Objective: To systematically evaluate the clinical effect of Xiaoke Decoction in the treatment of type 2 diabetes. Methods: Chinese databases such as CNKI, Wanfang, Weipu Chinese Biomedical Journal Database, and Chinese Medical Biological Literature Database, PubMed, Cochrane Library, Embase, and Web of Science were searched for English language literature from their inception until November 2019. A Meta-analysis was performed using RevMan 5.3 and Stata 12.0. Results: Thirty-eight studies were included in this study, with a total of 3757 patients. It was found that adding Xiaoke Decoction could improve total efficiency. The Xiaoke Decoction groups surpassed the western medicine groups regarding improvement in total efficiency (OR = 3.49; 95% CI: 2.78-4.39, P < 0.00001). Adding Xiaoke Decoction could lower the fasting plasma glucose (FPG) level. The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in FPG levels (MD = -1.14; 95% CI: -1.36 to 0.92, P < 0.00001). Adding Xiaoke Decoction could lower the 2 h postprandial blood glucose (2hPG) level. The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in 2hPG (MD = -1.40; 95% CI: -1.61 to 1.19, P < 0.00001). Adding Xiaoke Decoction could lower glycated hemoglobin (HbA1c). The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in HbA1c (MD = -0.77; 95% CI: -0.95 to 0.58, P < 0.00001). It was found that adding Xiaoke Decoction could lower the traditional Chinese medicine (TCM) syndrome score. The TCM syndrome scores among patients in the Xiaoke Decoction group were lower than those among patients in the control group after treatment (MD = -4.90; 95% CI: -7.22 to 2.57, P < 0.0001). At the same time, we conducted a subgroup and sensitivity analysis of age and intervention duration on the heterogeneity of total efficiency, FPG, 2hPG, HbAlc, and TCM syndrome score outcome indicators. For detecting publication bias, an egger test was conducted. Conclusion: Compared with western medicine alone, Xiaoke Decoction has more advantages for the treatment of type 2 diabetes with respect to total efficiency, FPG, 2hPG, HbAlc, and TCM syndrome score.

The introduction of an N-glycosylation site into prochymosin greatly enhances its production and secretion by Pichia pastoris.

BACKGROUND: N-glycosylation is one of the most important post-translational modifications. Many studies have shown that N-glycosylation has a significant effect on the secretion level of heterologous glycoproteins in yeast cells. However, there have been few studies reporting a clear and unified explanation for the intracellular mechanism that N-glycosylation affect the secretion of heterologous glycoproteins so far. Pichia pastoris is an important microbial cell factory producing heterologous protein. It is of great significance to study the effect of N-glycosylation on the secretion level of heterologous protein. Camel chymosin is a glycoprotein with higher application potential in cheese manufacturing industry. We have expressed camel prochymosin in P. pastoris GS115, but the lower secretion level limits its industrial application. This study attempts to increase the secretion level of prochymosin through N-glycosylation, and explore the molecular mechanism of N-glycosylation affecting secretion. RESULTS: Adding an N-glycosylation site at the 34th amino acid of the propeptide of prochymosin significantly increased its secretion in P. pastoris. N-glycosylation improved the thermostability of prochymosin without affecting the enzymatic activity. Immunoprecipitation coupled to mass spectrometry (IP-MS) analysis showed that compared with the wild prochymosin (chy), the number of proteins interacting with N-glycosylated mutant (chy34) decreased, and all differential interacting proteins (DIPs) were down-regulated in chy34-GS115 cell. The DIPs in endoplasmic reticulum were mainly concentrated in the misfolded protein pathway. Among the five DIPs in this pathway, overexpression of BiP significantly increased the secretion of chy. The knockout of the possible misfolded protein recognition elements, UDP-glycose:glycoprotein glucosyltransferase 1 and 2 (UGGT1/2) had no effect on the growth of yeast cells and the secretion of prochymosin. CONCLUSIONS: In conclusion, N-glycosylation increased the secretion of prochymosin in P. pastoris trough the adjustment of intracellular interacted proteins. The results of our study may help to elucidate the molecular mechanism of N-glycosylation affecting secretion and provide a new research method to improve the secretion of heterologous glycoprotein in P. pastoris.

Ectopic Overexpression of CsECR From Navel Orange Increases Cuticular Wax Accumulation in Tomato and Enhances Its Tolerance to Drought Stress.

Drought stress often occurred in citrus to limit its growth, distribution, and fruit quality. Cuticular waxes play an important role in regulating plant tolerance to drought stress. Plant enoyl-CoA reductase (ECR) is involved in the biosynthesis of cuticular waxes and catalyzes the last step of very long-chain fatty acids (VLCFAs) elongation. In this study, a putative ECR gene, named CsECR, was cloned from "Newhall" navel orange. CsECR protein has high identities with other plant ECR proteins and contained a conserved NADP/NAD-binding motif and three conserved functional sites. The highest expression of CsECR was observed in leaves, followed by stems, flavedos, ovaries, juice sacs, stigmas, stamens, albedos, and petals. Besides, the expression of CsECR was significantly induced by PEG6000 and ABA treatments. Ectopic overexpression of CsECR increased the contents of total waxes and aliphatic wax fractions (n-fatty acids, unsaturated fatty acids, n-alkanes, alkenes, iso-, and anteiso-alkanes) in the leaves and fruits of the transgenic tomato. Furthermore, ectopic overexpression of CsECR reduced the cuticle permeability in the leaves and fruits of the transgenic tomato and increased its tolerance to drought stress. Taken together, our results revealed that CsECR plays an important role in plant response to drought stresses by regulating cuticular wax biosynthesis.

Efficacy and safety of micronized purified flavonoid fractions for the treatment of postoperative hemorrhoid complications: A systematic review and meta-analysis.

BACKGROUND: Hemorrhoids, a common anorectal disease, seriously affects patients' quality of life. Micronized purified flavonoid fractions (MPFF) have been shown to improve hemorrhoid symptoms. PURPOSE: To evaluate the efficacy and safety of MPFF in treating postoperative hemorrhoid complications. STUDY DESIGN: A systematic review and meta-analysis of existing literature on natural compounds for treating postoperative complications of hemorrhoids. METHODS: A literature search was conducted using five databases, namely PubMed, WanFang, CNKI, Embase, and the Cochrane Library, to identify randomized controlled trials (RCTs) on the effects of MPFF treatment on hemorrhoids. Stata 15.1 and Revman 15.4 were used to assess the data, while subgroup and sensitivity analyses were performed to evaluate potential heterogeneity, and trial sequential analysis (TSA) and Egger test were used to evaluate the reliability of each trial. RESULTS: A total of 22 RCTs, including 2,335 participants were included in the analysis. MPFF improved the clinical efficacy of post-hemorrhoidectomy and reduced the bleeding rate, pain score, and edema score, although no substantial effect on adverse reactions was reported. Subgroup analyses showed a significant reduction in pain score and bleeding rate in trials with duration of 4-10 days and an improvement in clinical efficacy. Treatment for ≥ 10 days significantly improved the edema score; a dosage range of 1,800-2,700 mg/day of MPFF significantly reduced edema and pain scores, whereas < 1,800 mg/day significantly improved clinical efficacy. CONCLUSIONS: Based on searching the relevant literatures, this is the first meta-analysis on MPFF treatment of postoperative hemorrhoid complications. Our findings, validated by TSA, suggest that MPFF is safe and effective in reducing postoperative hemorrhoid complications, and that dose and duration are key factors in its efficacy, as illustrated by subgroup analysis. However, due to the small sample size, the standardized treatment regimen of MPFF could not be obtained; therefore, further research is warranted.

Diammonium Glycyrrhizinate Preparation for Liver Function Recovery in Chronic Hepatitis B in China: A Meta-analysis with Trial Sequential Analysis.

BACKGROUND: The treatment of chronic hepatitis B (CHB) comprises a global medical problem, and the first-line clinical drugs have obvious shortcomings. The use of the plant extract diammonium glycyrrhizinate (DG) in food and medicine has gradually widened because of its safety and effectiveness. DG is mainly used for liver-disease treatment in clinical practice, but DG intervention for CHB lacks systematic evidence. METHODS: The included randomized controlled trials were analyzed by comparator and control respectively for alanine aminotransferase (ALT), aspartate transaminase (AST), total bilirubin (TBIL) levels, hepatitis B virus DNA negative conversion ratio, and total effective rate, and subgroup analysis was conducted for intervention time, intervention dosage form, comparator drug, and combination drug, among others. Trial sequential analysis was used to verify the results. RESULT: DG could effectively reduce ALT, AST, TBIL, and other liver-function indexes and had a definite effect on liver-function recovery. From the beginning of the intervention to 3 months, the effect was significantly better than that of conventional treatment. Compared with other drugs, different dosage forms had differences in efficacy, and DG enteric-coated capsules and injections were lower than compound glycyrrhizin and magnesium isoglycyrrhizin. Meanwhile, DG capsules had no significant difference from them. Meanwhile, trial sequential analysis of the main results confirmed the reliability of the conclusion. CONCLUSION: To our knowledge, this was the first relatively complete meta-analysis and systematic evaluation of the efficacy of DG intervention for CHB; liver-function recovery was discussed in the context of traditional Chinese medicine thinking, and DG's therapeutic effect on CHB was defined.

The Quality Monitoring of Cistanches Herba (Cistanche deserticola Ma): A Value Chain Perspective.

Cistanche deserticola Ma was used as a medicine food homology, which was mainly produced in the Alxa region of northwest China. In recent years, it has been widely used in various food items. The increasing demand for Cistanches Herba has led to problems such as overexploitation and quality deterioration. The quality and safety of herbal medicines are critical and have been shown to be affected by the value chain (VC). Using the VC framework, the study is embedded in a larger study aiming to investigate the effects of different VCs types on the quality and stakeholders of Cistanches Herba. In this study, 90 Cistanches Herba samples were collected during fieldwork. An additional 40 samples were obtained from the herbal markets and medicine purchasing stations. Semi-structured interviews and key informant interviews were performed to collect data on stakeholders in major production areas. These samples were analyzed using high performance liquid chromatography (HPLC) coupled with the k-means clustering method; a targeted quality assessment strategy based on chemical analysis was adopted to understand the quality of Cistanches Herba. Based on market research, the collected samples were divided into different grades through k-means clustering analysis. Moreover, quality differences of Cistanches Herba in Alxa region were explored through DNA barcoding and chemical analysis. Accordingly, 10 different types of VCs were determined in the production of Cistanches Herba. The results show that there is a close relationship between the quality of Cistanches Herba and stakeholder benefits. Vertical integration at different levels was found for independent farmer-based VCs, horizontal collaboration was found in the cooperative-based VCs. The vertical coordination has led to a more consistent traceability system and strict regulation of supply chains. At the same time, the Cistanches Herba were divided into three grades. Through DNA barcoding and chemical analysis, we found that the quality differences between Cistanches Herba in the Alxa area were not significant. It was found that geographical suitability and vertical integration could impact the quality and sustainable production of Cistanches Herba. At the same time, the well-developed VCs can provide products with reliable quality, and ensure adequate financial revenue for relevant stakeholders.

Comparative analysis of the cuticular waxes and related gene expression between 'Newhall' and 'Ganqi 3' navel orange during long-term cold storage.

Previously, we obtained a wax-deficient mutant 'Ganqi 3' (MT) from 'Newhall' navel orange (Citrus sinensis [L.] Osbeck cv. Newhall, WT). The weight loss and postharvest decay in MT fruit were much higher than those in WT fruit after long-term cold storage. To understand the underlying mechanism, the changes in the morphology, chemical composition and gene expression of cuticular waxes between WT and MT fruit were compared during 150 days of storage at 4 °C. The density of epicuticular wax crystals and the contents of most of the aliphatic wax fractions in MT fruit were much lower than those in WT fruit over 90 days of storage. Further research revealed that the differences in the morphology and chemical composition of cuticular waxes might be important causes for the differences of postharvest weight loss and decay rates between WT and MT fruit. Notably, the expression profiles of 16 wax-related genes in WT and MT fruit were consistent with the change trends of corresponding cuticular wax components during cold storage. These results suggest that the morphology and chemical composition of cuticular waxes may be regulated by wax-related genes and play an important role in regulating the postharvest weight loss and the tolerances to postharvest decay in navel orange.

Quality Control of Glehniae Radix, the Root of Glehnia Littoralis Fr. Schmidt ex Miq., Along its Value Chains.

Glehniae Radix (GR) is one of the major medicinal materials in China. The global demand for GR, especially in Asian countries, is constantly increasing, and the supply of wild medicinal materials falls short of the demand. Previous studies have shown that the production and processing modes of different value chains (VCs) impact the quality of medicinal materials. After 4 years of field and market research, this study includes interviews with stakeholders in the VCs, integrates different types of VCs, and further analyzes the VCs. GR characteristics were also assessed; the length and upper-middle diameter of the collected samples were measured, and the effective components of the samples were determined to rank the GR samples according to their quality. The effective components were further analyzed by the K-means clustering method. Concomitantly, the local price (the sales price of the place where the medicinal materials are produced) and market price (the sale price of medicinal materials in the market) of GR in Chifeng, Inner Mongolia, and Anguo, Hebei, were documented, and the ARIMA (Autoregressive Integrated Moving Average) method was used to predict the GR price. Ten VCs are summarized in this article. The results showed that the income of the staff at the beginning of the VC is inadequate. Regarding GR origin, Inner Mongolia GR showed higher quality than that of other areas. As a result, the price of medicinal materials is relatively high, which corresponds to the market price of the survey. The forecast results showed that the market price of GR would increase slightly in the future, which could provide reference for the selection of medicinal materials cultivation in the future. Through the study, it was found that the vertical integration in the VCs of GR could guarantee not only the benefit of the growers but also the traceability of the medicinal materials, which further guarantees the quality of the medicinal materials. However, the complex relationship between the cultivation area and the quality of the medicinal materials is not clear, which should be addressed in future research.

The effect of air temperature on hospital admission of adults with community acquired pneumonia in Baotou, China.

The relationship between air temperature and the hospital admission of adult patients with community-acquired pneumonia (CAP) was analyzed. The hospitalization data pertaining to adult CAP patients (age ≥ 18 years) in two tertiary comprehensive hospitals in Baotou, Inner Mongolia Autonomous Region, China from 2014 to 2018 and meteorological data there in the corresponding period were collected. The exposure-response relationship between the daily average temperature and the hospital admission of adult CAP patients was quantified by using a distributed lag non-linear model. A total of 4466 cases of adult patients with CAP were admitted. After eliminating some confounding factors such as relative humidity, wind speed, air pressure, long-term trend, and seasonal trend, a lower temperature was found to be associated with a higher risk of adult CAP. Compared to 21 °C, lower temperature range of 4 to -12 °C was associated with a greater number of CAP hospitalizations among those aged ≥ 65 years, and the highest relative risk (RR) was 2.80 (95% CI 1.15-6.80) at a temperature of - 10 °C. For those < 65 years, lower temperature was not related to CAP hospitalizations. Cumulative lag RRs of low temperature with CAP hospitalizations indicate that the risk associated with colder temperatures appeared at a lag of 0-7 days. For those ≥ 65 years, the cumulative RR of CAP hospitalizations over lagging days 0-5 was 1.89 (95% CI 1.01-3. 56). In brief, the lower temperature had age-specific effects on CAP hospitalizations in Baotou, China, especially among those aged ≥ 65 years.

Chitosan modifies glycemic levels in people with metabolic syndrome and related disorders: meta-analysis with trial sequential analysis.

BACKGROUND: Chitosan supplementation has been shown to modulate glycemic levels; however, studies have reported conflicting results. The present meta-analysis with trial sequential analysis was conducted to verify the overall influence of chitosan on glycemic levels in patients with metabolic syndrome. METHODS: The PubMed, Cochrane library, and EMBASE databases were systematically searched for randomized controlled studies of chitosan intake and glycemic levels. RESULTS: A total of ten clinical trials including 1473 subjects were included in this meta-analysis. Pooled effect sizes were determined by random-effects meta-analysis. Subgroup analysis was performed to analyze the sources of heterogeneity and their influence on the overall results. The results revealed a significant reduction in fasting glucose levels (SMD: - 0.39 mmol/L, 95% CI: - 0.62 to - 0.16) and hemoglobin A1c (HbA1c) levels (SMD: -1.10; 95% CI: - 2.15 to - 0.06) following chitosan supplementation but no effect on insulin levels (SMD: - 0.20 pmol/L, 95% CI: - 0.64 to 0.24). Subgroup analyses further demonstrated significant reductions in fasting glucose levels in subjects administered 1.6-3 g of chitosan per day and in studies longer than 13 weeks. Trial sequential analysis of the pooled results of the hypoglycemic effect demonstrated that the cumulative Z-curve crossed both the conventional boundary and trial sequential monitoring boundary for glucose and HbA1c. CONCLUSIONS: The glucose level of patients who are diabetic and obese/overweight can be improved by supplementation with chitosan for at least 13 weeks at 1.6-3 g per day. Additional clinical research data are needed to confirm the role of chitosan, particularly in regulating glycosylated hemoglobin and insulin.

Ectopic Expression of CsKCS6 From Navel Orange Promotes the Production of Very-Long-Chain Fatty Acids (VLCFAs) and Increases the Abiotic Stress Tolerance of Arabidopsis thaliana.

Cuticular wax is closely related to plant resistance to abiotic stress. 3-Ketoacyl-CoA synthase (KCS) catalyzes the biosynthesis of very-long-chain fatty acid (VLCFA) wax precursors. In this study, a novel KCS family gene was isolated from Newhall navel orange and subsequently named CsKCS6. The CsKCS6 protein has two main domains that belong to the thiolase-like superfamily, the FAE1-CUT1-RppA and ACP_syn_III_C domains, which exist at amino acid positions 80-368 and 384-466, respectively. CsKCS6 was expressed in all tissues, with the highest expression detected in the stigma; in addition, the transcription of CsKCS6 was changed in response to drought stress, salt stress and abscisic acid (ABA) treatment. Heterologous expression of CsKCS6 in Arabidopsis significantly increased the amount of VLCFAs in the cuticular wax on the stems and leaves, but there were no significant changes in total wax content. Compared with that of the wild-type (WT) plants, the leaf permeability of the transgenic plants was lower. Further research showed that, compared with the WT plants, the transgenic lines experienced less water loss and ion leakage after dehydration stress, displayed increased survival under drought stress treatment and presented significantly longer root lengths and survival under salt stress treatment. Our results indicate that CsKCS6 not only plays an important role in the synthesis of fatty acid precursors involved in wax synthesis but also enhances the tolerance of transgenic Arabidopsis plants to abiotic stress. Thus, the identification of CsKSC6 could help to increase the abiotic stress tolerance of Citrus in future breeding programs.

[Study on traditional processing method of Mongolian medicine and excipient usage based on data mining].

This paper explores Mongolian medicine processing methods and the use regularity of excipient by text mining techniques. Relevant books of Mongolian medicine processing were consulted to collect data on Mongolian medicine processing methods and excipient, and select data based on processing methods and excipient noun frequency statistics. Microsoft Excel 2010 software was used for statistical analysis and mining for the usage regularity of different types of Mongolian medicinal materials in different periods. And Cytoscape 3.6.1 software was used for visual presentation. The topological analysis showed the top five processing methods were net production, development, frying, calcining and cooking, and the top five processing excipient were fresh milk, wine, urine, cream and mineral borax. Frequency analysis showed that the plant medicinal materials were mostly recorded in the 18~(th) and 21~(st) centuries, especially in the 21 st century; the processing methods mostly contained water processing, repair processing and other methods. The mineral medicinal materials were mostly recorded in the 18~(th), 19~(th) and 21~(st) centuries; most of the processing methods were the fire processing method. The animal medicinal materials were recorded in the 18~(th), 19~(th) and 21~(st) century; the fire processing method occupied a major position, and the repair processing and the grinding processing were markedly increased in the 21~(st) century. In the use of excipient, liquid excipient were mostly used in plant medicines. Solid excipient were most commonly used in the 18~(th) century. Animal excipient were mostly used during the processing in the 18~(th) century. The use of liquid excipient gradually increased in the 19~(th) and 21~(st) centuries. This study summarizes the traditional processing methods of Mongolian medicine and the usage regularity of excipient, defines the characteristics of Mongolian medicine processing methods and excipient, and the characteristics of the combination of medicinal materials and excipient, so as to provide reference for the clinical use of Mongolian medicine.

A Na+/H+ antiporter, K2-NhaD, improves salt and drought tolerance in cotton (Gossypium hirsutum L.).

KEY MESSAGE: Overexpression of K2-NhaD in transgenic cotton resulted in phenotypes with strong salinity and drought tolerance in greenhouse and field experiments, increased expression of stress-related genes, and improved regulation of metabolic pathways, such as the SOS pathway. Drought and salinity are major abiotic stressors which negatively impact cotton yield under field conditions. Here, a plasma membrane Na+/H+ antiporter gene, K2-NhaD, was introduced into upland cotton R15 using an Agrobacterium tumefaciens-mediated transformation system. Homozygous transgenic lines K9, K17, and K22 were identified by PCR and glyphosate-resistance. TAIL-PCR confirmed that T-DNA carrying the K2-NhaD gene in transgenic lines K9, K17 and K22 was inserted into chromosome 3, 19 and 12 of the cotton genome, respectively. Overexpression of K2-NhaD in transgenic cotton plants grown in greenhouse conditions and subjected to drought and salinity stress resulted in significantly higher relative water content, chlorophyll, soluble sugar, proline levels, and SOD, CAT, and POD activity, relative to non-transgenic plants. The expression of stress-related genes was significantly upregulated, and this resulted in improved regulation of metabolic pathways, such as the salt overly sensitive pathway. K2-NhaD transgenic plants growing under field conditions displayed strong salinity and drought tolerance, especially at high levels of soil salinity and drought. Seed cotton yields in transgenic line were significantly higher than in wild-type plants. In conclusion, the data indicate that K2-NhaD transgenic lines have great potential for the production of stress-tolerant cotton under field conditions.

[Application and analysis of ethnic medicines in "Three Minorities" in Inner Mongolia].

This paper investigates and counts the ethnic medicines of the Ewenki,Daur and Oroqen ethnic groups,which are known as the " Three Minorities" in Inner Mongolia. Through the methods of literature collection,interview investigation,and resource investigation,different ethnic medicines were collected on the main diseases,drug varieties,drug-injection sites,and drug administration methods. Through data statistics and SPSS analysis,the similarities and individual differences between the three ethnic groups were clarified. The results indicated the predicament of the current national medicine,which is helpful for the protection and inheritance of ethnic medicine.