Effects of butylphthalide combined with hyperbaric oxygen therapy on post-stroke cognitive impairment / 中国基层医药

Objective:To investigate the clinical efficacy of butylphthalide combined with hyperbaric oxygen therapy on post-stroke cognitive impairment in patients with acute ischemic stroke.Methods:A total of 90 patients with post-stroke cognitive impairment who were hospitalized within 72 hours of onset in Suining County People's Hospital from December 2019 to November 2020 were included in this study. They were randomly divided into a control group and an observation group ( n = 45/group). The control group was given conventional treatment and the observation group was given butylphthalide combined with hyperbaric oxygen therapy in addition to conventional treatment. The National Institutes of Health Stroke Scale score, Montreal Cognitive Assessment score, and Activities of Daily Living score were compared between the two groups before and after treatment. Results:Before treatment, there were no significant differences in the National Institutes of Health Stroke Scale score, Montreal Cognitive Assessment score, and Activities of Daily Living score between the two groups (all P > 0.05). At 14 days and 1 month after surgery, the National Institutes of Health Stroke Scale scores in the observation group were (4.02 ± 2.18) points and (3.21 ± 2.03) points, which were significantly lower than (5.21 ± 2.24) points and (4.62 ± 2.68) points in the control group ( t =2.55, 2.81, both P < 0.05). At 1 and 3 months after treatment, the Montreal Cognitive Assessment score in the observation group were (19.79 ± 5.67) points and (23.69 ± 2.67) points, which were significantly higher than (16.88 ± 5.12) points and (19.74 ± 2.29) points in the control group ( t = 2.56, 7.53, both P < 0.05). At 1 and 3 months after treatment, Activities of Daily Living scores in the observation group were (54.85 ± 5.69) points and (74.38 ± 4.98) points, which were significantly higher than (46.78 ± 6.24) points and (63.21 ± 5.24) points in the control group ( t = 6.41, 9.76, both P < 0.05). Conclusion:Butylphthalide combined with hyperbaric oxygen therapy for the treatment of post-stroke cognitive impairment in patients with acute ischemic stroke can alleviate neurologic deficits, and improve cognitive function and the ability of daily life.

Feasibility study on local infiltration anesthesia for postoperative analgesia in patients with hallux valgus / 中国骨伤

OBJECTIVE@#To evaluate the clinical efficacy of local infiltration anesthesia of ropivacaine combined with compound betamethasone for postoperative analgesia in patients with hallux valgus.@*METHODS@#From September 2019 to December 2020, 48 patients with hallux valgus were treated surgically. According to different postoperative analgesia methods, the patients were divided into combined local infiltration group and intravenous analgesia pump group. There were 24 cases, in the combined local infiltration group including 2 males and 22 females;the age ranged from 21 to 78 years old, with an average of (58.3±7.7) years old;soft tissue release and chevron osteotomy were performed in 15 cases and metatarsophalangeal joint fusion in 9 cases;immediately after operation, 20 ml of ropivacaine combined with compound betamethasone mixed diluent was used for local infiltration anesthesia once. There were 24 patients in intravenous analgesia pump group, including 3 males and 21 females;the age ranged from 23 to 81 years old, with an average of(56.8±8.3) years old;soft tissue release and Chevron osteotomy were performed in 17 cases and metatarsophalangeal joint fusion in 7 cases;immediately after operation, intravenous analgesia pump was used for analgesia. The basic flow was 2 ml / h;the self control dose was 0.5 ml;and the locking time was 15 min. Visual analogue scale (VAS) was recorded at 12, 24, 48 and 72 hours after operation;and the VAS was recorded at 24 hours after operation. The occurrence of adverse drug reactions at 0 to 12 hours, 12 to 24 hours and 24 to 48 hours after operation were recorded;and the healing of incision was recorded.@*RESULTS@#All patients were followed up, and the duration ranged from 14 to 17 days, with a mean of (14.60±0.92) days. There was significantdifference in VAS at 12, 24 and 48 hours between the combined local infiltration group and the intravenous analgesia pump group(@*CONCLUSION@#Compared with intravenous analgesia pump group, ropivacaine combined with compound betamethasone can significantly reduce postoperative wound pain without increasing adverse drug reactions, and does not increase wound infection.

L-theanine: an astounding sui generis amino acid in poultry nutrition.

L-theanine (γ-Glutamylethylamide) is a nonprotein water soluble amino acid (AA) mostly found in leaves of Camellia sinensis (green tea). This is a key component of green tea and is considered as the most abundant form of total AAs in green tea (i.e., about 50%). L-theanine is an exclusive taste ingredient of tea producing an attractive flavor and aroma in tea. It has biological effects such as antioxidant, growth promoter, immune booster, anti-stresser, hepatoprotective, antitumor, antiaging, antimicrobial, anti-inflammatory, and antianxiety activities that are worth noticing. It could reduce the oxidative impairment by reducing the synthesis of reactive oxygen species, oxidative parameters, and lipid damage as well as increasing the activity of antioxidant enzymes. The oral ingestion of L-theanine enhanced γδ T-cell proliferation. Therefore, it is being considered an essential compound of green tea that has the ability to improve immune function. The L-theanine can be used as a potential treatment for hepatic injury and immune-related liver diseases via the downregulation of the inflammatory response through the initiation of nitric oxide synthesis and glutathione production which are likely to be critical for the control of hepatic diseases as well as for the improvement of immune function. In addition, it could be used as a best natural feed additive with a potent antistressor by decreasing the levels of corticosterone, dopamine, and noradrenaline. After systematically reviewing the literature, it is noticed that most studies were carried out on mice, pig, human, and butterfly; while dietary supplementation studies of L-theanine in animal and poultry especially among broilers are very limited because of less awareness of this AA. So, the aim of this review is to encourage the veterinarian and poultry researchers to conduct more research at the molecular level about this AA to expose its more beneficial effects and its mechanism of absorption for potential use of this unique green tea AA in poultry nutrition.

Using Guduchi (Tinospora cordifolia) as an eco-friendly feed supplement in human and poultry nutrition.

Guduchi (Tinospora cordifolia) is a well-recognized and widely distributed traditional plant that is used successfully in Indian Ayurveda medicine. T. cordifolia has shown many promising biological activities, such as antioxidative, antimicrobial, antihyperglycemic, anti-inflammatory, osteoprotective, hepatoprotective, antidiarrheal, and antistress effects. Guduchi is a rich source of protein and micronutrients, such as iron, zinc, copper, calcium, phosphorus, and manganese. It also contains many secondary plant metabolites, such as terpenes, alkaloids, flavonoids, steroids, and glycosides. Based on previous studies in poultry, the supplementation levels of Guduchi range from 1 to 5 g/kg of diet (different sources, such as powder, extracts, roots, and leaves, have been used). It was suggested that this variation in supplementation levels depends on different factors, including the extraction method, the supplementation proposed, the method of supplementation (either in feed or drinking water), and the species and physiological status of the birds. Generally, dietary supplementation of poultry broilers with T. cordifolia yielded positive impacts on growth performance, body gains (increased by 4.8%), dressing percentage (increased by 7.1%), meat quality traits, and the shelf life of the meat. In addition, T. cordifolia exerted a palliative effect on the general health status of the birds through reducing live enzymes and plasma uric acids and enhancing the immune response, as indicated by the leukocyte count, hemagglutinin titer, interleukin activity, and mortality levels. Further investigations concluded that T. cordifolia showed strong antimicrobial effects against Escherichia coli and Salmonella enteritidis, with subsequent reductions in mortality. Moreover, T. cordifolia showed an ability to improve humoral and cell-mediated immunity against Newcastle disease, infectious anemia, gout, and aflatoxicosis. The current review discusses many beneficial properties of T. cordifolia, although the lack of pharmacological trials limits the use of this extract in poultry. Further research should be performed regarding the composition of the active compound, the possible mechanisms of action, and the effective doses to fully understand the activities and benefits of T. cordifolia as a growth performance improvement supplement.

Molecular genetics of medicinal plants, past achievement, and new era / 中国中药杂志

Unraveling the genetic basis of medicinal plant metabolism and developmental traits is a long-standing goal for pharmacologists and plant biologists. This paper discusses the definition of molecular genetics of medicinal plants, which is an integrative discipline with medicinal plants as the research object. This discipline focuses on the heredity and variation of medicinal plants, and elucidates the relationship between the key traits of medicinal plants(active compounds, yield, resistance, etc.) and genotype, studies the structure and function, heredity and variation of medicinal plant genes mainly at molecular level, so as to reveal the molecular mechanisms of transmission, expression and regulation of genetic information of medicinal plants. Specifically, we emphasize on three major aspects of this discipline.(1)Individual and population genetics of medicinal plants, this part mainly highlights the genetic mechanism of the domestication, the individual genomics at the species level, and the formation of genetic diversity of medicinal plants.(2)Elucidation of biosynthetic pathways of active compounds and their evolutionary significance. This part summarizes the biosynthesis, diversity and molecular evolution of active compounds in medicinal plants.(3) Molecular mechanisms that shaping the key agronomic traits by internal and external factors. This part focuses on the accumulation and distribution of active compounds within plants and the regulation of metabolic network by environmental factors. Finally, we prospect the future direction of molecular genetics of medicinal plants based on the rapid development of multi-omics technology, as well as the application of molecular genetics in the future strategies to achieve conservation and breeding of medicinal plants and efficient biosynthesis of active compounds.

In ovo delivery of various biological supplements, vaccines and drugs in poultry: current knowledge.

The technique of delivering various nutrients, supplements, immunostimulants, vaccines, and drugs via the in ovo route is gaining wide attention among researchers worldwide for boosting production performance, immunity and safeguarding the health of poultry. It involves direct administration of the nutrients and biologics into poultry eggs during the incubation period and before the chicks hatch out. In ovo delivery of nutrients has been found to be more effective than post-hatch administration in poultry production. The supplementation of feed additives, nutrients, hormones, probiotics, prebiotics, or their combination via in ovo techniques has shown diverse advantages for poultry products, such as improved growth performance and feed conversion efficiency, optimum development of the gastrointestinal tract, enhancing carcass yield, decreased embryo mortality, and enhanced immunity of poultry. In ovo delivery of vaccination has yielded a better response against various poultry pathogens than vaccination after hatch. So, this review has aimed to provide an insight on in ovo technology and its potential applications in poultry production to deliver different nutrients, supplements, beneficial microbes, vaccines, and drugs directly into the developing embryo to achieve an improvement in post-hatch growth, immunity, and health of poultry. © 2019 Society of Chemical Industry.

Potential nutraceutical and food additive properties and risks of coffee: a comprehensive overview.

Coffee is a composite mixture of more than a thousand diverse phytochemicals like alkaloids, phenolic compounds, vitamins, carbohydrates, lipids, minerals and nitrogenous compounds. Coffee has multifunctional properties as a food additive and nutraceutical. As a nutraceutical, coffee has anti-inflammatory, anti-oxidant, antidyslipidemic, anti-obesity, type-2 diabetes mellitus (DM), and cardiovascular diseases (CVD), which can serve for the treatment and prevention of metabolic syndrome and associated disorders. On the other hand, as a food additive, coffee has antimicrobial activity against a wide range of microorganisms, inhibits lipid peroxidation (LPO), and can function as a prebiotic. The outcomes of different studies also revealed that coffee intake may reduce the incidence of numerous chronic diseases, like liver disease, mental health, and it also overcomes the all-cause mortality, and suicidal risks. In some studies, high intake of coffee is linked to increase CVD risk factors, like cholesterol, plasma homocysteine and blood pressure (BP). There is also a little evidence that associated the coffee consumption with increased risk of lung tumors in smokers. Among adults who consume the moderate amount of coffee, there is slight indication of health hazards with strong indicators of health benefits. Moreover, existing literature suggests that it may be cautious for pregnant women to eliminate the chances of miscarriages and impaired fetal growth. The primary purpose of this narrative review is to provide an overview of the findings of the positive impacts and risks of coffee consumption on human health. In conclusion, to date, the best available evidence from research indicates that drinking coffee up to 3-4 cups/day provides health benefits for most people.

16S ribosomal RNA sequencing reveals a modulation of intestinal microbiome and immune response by dietary L-theanine supplementation in broiler chickens.

Despite the availability of abundant literature on green tea, studies on the use of L-theanine (an amino acid found only in green tea) as a feed additive in poultry especially broiler are limited. So, this study was conducted to explore the effects of L-theanine on the intestinal microbiome and immune response in a broiler. A total of 400-d-old chicks were randomly divided into four treatment groups (A, B, C, and D) using a complete randomized design. Treatments were as follows: A, control (basal diet); B, basal diet + 100 mg L-theanine/kg diet; C, basal diet + 200 mg L-theanine/kg diet; and D, basal diet + 300 mg L-theanine/kg diet. Mucosal samples from ileum and jejunum of broiler chicken were extracted at 21 and 42 d of age. Extraction of genomic DNA was followed by amplification of V3 and V4 hypervariable regions of 16S ribosomal RNA. After Illumina sequencing, results revealed that treatment with L-theanine significantly increased the population of Lactobacillus in ileum and jejunum as compared to a control group, but the higher population was observed in jejunum at both 21 and 42 d of age. The overall diversity of the jejunum microbiome in the treatment group was significantly lower than that of the ileum and control group (P < 0.05). Results of this study revealed that mRNA expression of TLRs (TLR-2 and TLR-4) and cytokines (TNF-α, IFN-γ, and IL-2) was decreased in response to treatment with L-theanine. Moreover, the negative correlation of abundance of Lactobacillus was observed with expression of IL-2 and IFN-γ in the intestine and these effects were highly significant (P < 0.01). In summary, our finding revealed that dietary supplementation of L-theanine exhibited a positive influence on intestinal bacteria by supporting beneficial microbes like Lactobacillus while decreasing harmful microbes like Clostridium.

Yucca schidigera can mitigate ammonia emissions from manure and promote poultry health and production.

Mojave yucca (Yucca schidigera) is widely grown in the deserts. This herb is commercially used because it is rich in saponins and phenolic compounds with antioxidant effect. Y. schidigera or its derivatives are included as nontoxic food supplements, in cosmetics, and in the pharmaceutical industry. Saponins originated from Y. schidigera have anti-inflammatory, antioxidant, immunostimulatory, growth promoter, hypocholesterolemic, and hypoglycemic effects. To date, the key role of Y. schidigera or its products in animal nutrition is to reduce the ammonia content in the atmosphere and fecal odor in poultry excreta. Mitigating ammonia by using this plant could be achieved by the modification of gut microbiota, enhancement in digestion, and absorption of nutrients, leading to a better growth and production performance of animals and poultry. Various methods were applied to mitigate the emission of odor from the litter by different strategies including biofilters, litter treatments, air scrubbers, neutralizing agents, windbreak walls, etc., but these techniques are expensive. This article provides a new insight to scientists and poultry breeders to use Y. schidigera plant or its products as inexpensive and safe sources of a feed supplement to overcome the ammonia and fecal odor problems, as well as reduce environmental pollution in poultry houses.

Research advances in mechanism of active components of traditional Chinese medicine for reducing uric acid / 中国药理学通报

Hyperuricemia is a heterogeneous group of diseases caused by disorders of purine metabolism and /or excretion of uric acid reduction .Chinese medicine treatment of hyperurice-mia has gradually attracted attention , and made significant pro-gress.In recent years a large number of experimental studies have shown that flavonoids , saponins , alkaloids and other active components of traditional Chinese medicine can reduce uric acid by inhibiting the activity of key enzymes in purine metabolism ,or by increasing the excretion of uric acid through regulating the expression of uric acid transporters , thereby playing an important role in reducing uric acid .Based on the pathogenesis of hyperu-ricemia, the mechanism of active components of traditional Chi-nese medicine for reducing uric acid is summarized , and recent research reports are reviewed .

Reconsidering betaine as a natural anti-heat stress agent in poultry industry: a review.

Betaine is found ubiquitously in plants, animals, microorganisms, and rich dietary sources including seafood, spinach, and wheat bran. The chief physiological role of betaine is to function as a methyl donor and an osmolyte. Betaine also acts as an osmolyte, to maintain the avian's cellular water and ion balance to improve the avian's capacity against heat stress via preventing dehydration and osmotic inactivation. It helps in maintaining the protective osmolytic activity, especially in heat-stressed birds. Betaine may promote various intestinal microbes against osmotic variations and thus improve microbial fermentation activity. Previous studies showed that dietary supplementation of betaine in poultry diets could positively affect nutrients' digestibility, reduce abdominal fat weight, and increase breast meat yield. In addition, betaine has been reported to protect internal organs and boost their performance. Its inclusion in poultry diet is sparing essential amino acids like choline and methionine. In addition, it may play an important role in lean meat production by positively affecting the lipid metabolism with increased fatty acids catabolism and thus reducing carcass fat deposition. The aim of this review article was to broaden the knowledge regarding betaine and its importance in the poultry industry to cope with the heat stress problem. Moreover, it should be added to the diet as a natural anti-stressor through different routes (water/feed) to overcome the heat stress problem. However, further studies need to be conducted at the genetic and molecular basis to elucidate the mechanism behind the betaine as a natural anti-heat agent to decrease the heat stress problem in the poultry industry.

Effect of computer-assisted postural balance training on balance disorders after cerebellar stroke / 国际脑血管病杂志

Objective To investigate the efficacy of computer-assisted postural balance training system combined with Bobath balance training on balance disorders after cerebellar stroke.Methods Forty patients with balance disorders after cerebellar stroke were randomly divided into either a combined training group or a control group (n =20 in each group).The control group was trained with Bobath balance training and routine rehabilitation treatment.On the basis of this,the combined training group used the computerassisted postural balance training system for rehabilitation treatment.The balance function,activities of daily living and motor function were evaluated with the Berg Balance Scale (BBS),modified Barthel Index,mBI)and Fugl-Meyer Assessment (FMA) before and after treatment,and the computer-assisted postural balance training system was used to conduct the postural balance ability according to the center of gravity track length,rectangle area of track,and peripheral area of track.Results There were no significant differences in various indexes before treatment between the patients of both groups.The scores of BBS,mBI and FMA after treatment were increased obviously (all P ＜ 0.001).The the center of gravity track length,rectangle area of track,and peripheral area of track were decreased compared before treatment (P ＜0.001 or 0.05).In addition to the FMA score,all the indexes of the combined training group after treatment were significantly better than those of the control group (all P＜0.05).Conclusions On the basis of Bobath balance training,using computer-assisted postural balance training system can obviously improve the balance disorder and extremities motion ability after cerebellar stroke,and thus improving the activities of daily living in cerebellar stroke patients.

Advance in transcriptomic studies of ginseng species / 中国中药杂志

There are many valuable medicinal plants in Ginseng genus belonging to Araliaceae. Among them, Panax ginseng, P. quinquefolium and P. notoginseng are the most famous species. With the development of next-generation sequencing (NGS) technologies, sequencing and analysis of transcriptomes have become powerful tools for discovery of novel genes, screening molecular markers and elucidation of specific biosynthetic pathway of secondary metabolites. Their transcriptomes provided abundant genes for further study on functional genomics. Here this paper summarized the recent advances in the transcriptomic studies of these three medicinal plants, including discovery of novel genes and elucidation of metabolic regulation, which will contribute to functional genomics in ginseng species.

Advance in flavonoids biosynthetic pathway and synthetic biology / 中国中药杂志

Flavonoids are the valuable components in medicinal plants, which possess a variety of pharmacological activities, including anti-tumor, antioxidant and anti-inflammatory activities. There is an unambiguous understanding about flavonoids biosynthetic pathway, that is,2S-flavanones including naringenin and pinocembrin are the skeleton of other flavonoids and they can transform to other flavonoids through branched metabolic pathway. Elucidation of the flavonoids biosynthetic pathway lays a solid foundation for their synthetic biology. A few flavonoids have been produced in Escherichia coli or yeast with synthetic biological technologies, such as naringenin, pinocembrin and fisetin. Synthetic biology will provide a new way to get valuable flavonoids and promote the research and development of flavonoid drugs and health products, making flavonoids play more important roles in human diet and health.

Strategies of elucidation of biosynthetic pathways of natural products / 中国中药杂志

Elucidation of the biosynthetic pathways of natural products is not only the major goal of herb genomics, but also the solid foundation of synthetic biology of natural products. Here, this paper reviewed recent advance in this field and put forward strategies to elucidate the biosynthetic pathway of natural products. Firstly, a proposed biosynthetic pathway should be set up based on well-known knowledge about chemical reactions and information on the identified compounds, as well as studies with isotope tracer. Secondly, candidate genes possibly involved in the biosynthetic pathway were screened out by co-expression analysis and/or gene cluster mining. Lastly, all the candidate genes were heterologously expressed in the host and then the enzyme involved in the biosynthetic pathway was characterized by activity assay. Sometimes, the function of the enzyme in the original plant could be further studied by RNAi or VIGS technology. Understanding the biosynthetic pathways of natural products will contribute to supply of new leading compounds by synthetic biology and provide "functional marker" for herbal molecular breeding, thus but boosting the development of traditional Chinese medicine agriculture.

Metabolomics research of medicinal plants / 中国中药杂志

Metabolomics is the comprehensively study of chemical processes involving small molecule metabolites. It is an important part of systems biology, and is widely applied in complex traditional Chinese medicine（TCM）system. Metabolites biosynthesized by medicinal plants are the effective basis for TCM. Metabolomics studies of medicinal plants will usher in a new period of vigorous development with the implementation of Herb Genome Program and the development of TCM synthetic biology. This manuscript introduces the recent research progresses of metabolomics technology and the main research contents of metabolomics studies for medicinal plants, including identification and quality evaluation for medicinal plants, cultivars breeding, stress resistance, metabolic pathways, metabolic network, metabolic engineering and synthetic biology researches. The integration of genomics, transcriptomics and metabolomics approaches will finally lay foundation for breeding of medicinal plants, R&D, quality and safety evaluation of innovative drug.

Progress in the study of Velvet and LaeA proteins and their relation to the development and bioactive compounds in medicinal fungi / 药学学报

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.

Progress in the study of Velvet and LaeA proteins and their relation to the development and bioactive compounds in medicinal fungi / 药学学报

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.

Cloning and expression analysis of a key device of HMGR gene involved in ginsenoside biosynthesis of Panax ginseng via synthetic biology approach / 药学学报

3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR), the first enzyme of mavalonic acid pathway, is one of the key devices involved in ginsenoside biosynthesis based on synthetic biology approach. The open reading frame of a novel HMGR gene from Panax ginseng (PgHMGR2) was cloned and analyzed in this study. PgHMGR2-encoding protein showed 71.6% sequence similarity to a P. ginseng HMGR in GenBank. The full-length cDNA sequence of PgHMGR2 containing 1 770 bp, which encodes 589 amino acids, was cloned by RT-PCR strategy from P. ginseng. The bioinformatic analysis showed that PgHMGR2-encoding protein contained two transmembrane regions and the HMG_CoA_reductase domain, without signal peptide. The protein sequence of PgHMGR2 had the highest sequence similarity (99%) with Panax quinquefolius HMGR (GenBank accession No. ACV65036). The expression level of PgHMGR2 was the highest in flower based on a real-time PCR analysis, followed by leaf and root, and the lowest was in stem. The result will provide a foundation for exploring the molecular function of PgHMGR2 involved in ginsenoside biosynthesis based on synthetic biology approach in P. ginseng plants.

Development of the devices for synthetic biology of triterpene saponins at an early stage: cloning and expression profiling of squalene epoxidase genes in panax notoginseng / 药学学报

Synthetic biology of traditional Chinese medicine (TCM) is a new and developing subject based on the research of secondary metabolite biosynthesis for nature products. The early development of synthetic biology focused on the screening and modification of parts or devices, and establishment of standardized device libraries. Panax notoginseng (Burk.) F.H.Chen is one of the most famous medicinal plants in Panax species. Triterpene saponins have important pharmacological activities in P. notoginseng. Squalene epoxidase (SE) has been considered as a key rate-limiting enzyme in biosynthetic pathways of triterpene saponins and phytosterols. SE acts as one of necessary devices for biosynthesis of triterpene saponins and phytosterols in vitro via synthetic biology approach. Here we cloned two genes encoding squalene epoxidase (PnSE1 and PnSE2) and analyzed the predict amino acid sequences by bioinformatic analysis. Further, we detected the gene expression profiling in different organs and the expression level of SEs in leaves elicited by methyl jasmonate (MeJA) treatment in 4-year-old P notoginseng using real-time quantitative PCR (real-time PCR). The study will provide a foundation for discovery and modification of devices in previous research by TCM synthetic biology. PnSE1 and PnSE2 encoded predicted proteins of 537 and 545 amino acids, respectively. Two amino acid sequences predicted from PnSEs shared strong similarity (79%), but were highly divergent in N-terminal regions (the first 70 amino acids). The genes expression profiling detected by real-time PCR, PnSE1 mRNA abundantly accumulated in all organs, especially in flower. PnSE2 was only weakly expressed and preferentially in flower. MeJA treatment enhanced the accumulation of PnSEI mRNA expression level in leaves, while there is no obvious enhancement of PnSE2 in same condition. Results indicated that the gene expressions of PnSE1 and PnSE2 were differently transcribed in four organs, and two PnSEs differently responded to MeJA stimuli. It was strongly suggested that PnSEs play different roles in secondary metabolite biosynthesis in P. notoginseng. PnSE1 might be involved in triterpenoid biosynthesis and PnSE2 might be involved in phytosterol biosynthesis.