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BACKGROUND: Astaxanthin is a kind of tetraterpene and has strong antioxygenic property. The biosynthesis of astaxanthin in engineered microbial chassis has greater potential than its chemical synthesis and extraction from natural producers in an environmental-friendly way. However, the cost-offsetting production of astaxanthin in engineered microbes is still constrained by the poor efficiency of astaxanthin synthesis pathway as a heterologous pathway. RESULTS: To address the bottleneck of limited production of astaxanthin in microbes, we developed in vitro and in vivo recombination methods respectively in engineered yeast chassis to optimize the combination of heterologous ß-carotene ketolase (crtW) and hydroxylase (crtZ) modules that were selected from different species. As a result, the in vitro and in vivo recombination methods enhanced the astaxanthin yield respectively to 2.11-8.51 folds and 3.0-9.71 folds compared to the initial astaxanthin pathway, according to the different combination of particular genes. The highest astaxanthin producing strain yQDD022 was constructed by in vivo method and produced 6.05 mg g-1 DCW of astaxanthin. Moreover, it was proved that the in vivo recombination method showed higher DNA-assembling efficiency than the in vitro method and contributed to higher stability to the engineered yeast strains. CONCLUSIONS: The in vitro and in vivo recombination methods of heterologous modules provide simple and efficient ways to improve the astaxanthin yield in yeast. Both the two methods enable high-throughput screening of heterologous pathways through recombination of certain crtW and crtZ derived from different species. This study not only exploited the underlying optimal combination of crtZ and crtW for astaxanthin synthesis, but also provided a general approach to evolve a heterologous pathway for the enhanced accumulation of desired biochemical products.
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Vias Biossintéticas , Engenharia Metabólica/métodos , Recombinação Genética , Saccharomyces cerevisiae/metabolismo , Escherichia coli/metabolismo , Oxigenases de Função Mista/genética , Oxigenases/genética , Saccharomyces cerevisiae/genética , Xantofilas/metabolismoRESUMO
BACKGROUND: Pituitary stalk interruption syndrome (PSIS) is a rare anatomical defect of the pituitary gland falling under the spectrum of holoprosencephaly phenotypes. It is characterized by a deficiency in anterior pituitary hormones, such as growth hormone, gonadotropins, and thyroid hormones. Due to the syndrome's rarity and nonspecific manifestations, there is a lack of standardized treatment strategies. Consequently, early diagnosis through imaging and on-time intervention are crucial for improving patients' outcomes. CASE SUMMARY: A 30-year-old man presented with absent secondary sexual characteristics and azoospermia. Laboratory evaluation revealed a deficiency in gonadotropins, while thyroid function was mostly within normal ranges. Magnetic resonance imaging of the pituitary gland showed pituitary stalk agenesis, hypoplasia of the anterior pituitary, and ectopic posterior pituitary, leading to the diagnosis of PSIS. Initially, the patient underwent 6 mo of gonadotropin therapy without significant changes in hormone levels and secondary sexual characteristics. Pulsatile gonadotropin-releasing hormone therapy was then administered, resulting in the detection of sperm in the semen analysis within 3 mo. After 6 mo, routine semen tests showed normal semen quality. The couple faced challenges in conceiving due to abstinence and underwent three cycles of artificial insemination, which was unsuccessful. They also attempted in vitro fertilization, but unfortunately, the woman experienced a miscarriage 10 wk after the embryo transfer. CONCLUSION: Early detection, accurate diagnosis, and timely treatment are crucial in improving the quality of life and fertility of PSIS patients.
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Introduction: Soil salinization has become one of the most serious environmental issues globally. Excessive accumulation of soluble salts will adversely affect the survival, growth, and reproduction of plants. Elaeagnus angustifolia L., commonly known as oleaster or Russian olive, has the characteristics of tolerance to drought and salt. Arbuscular mycorrhizal (AM) fungi are considered to be bio-ameliorator of saline soils that can enhance the salt tolerance of the host plants. However, there is little information on the root proteomics of AM plants under salt stress. Methods: In this study, a label-free quantitative proteomics method was employed to identify the differentially abundant proteins in AM E. angustifolia seedlings under salt stress. Results: The results showed that a total of 170 proteins were significantly differentially regulated in E.angustifolia seedlings after AMF inoculation under salt stress. Mycorrhizal symbiosis helps the host plant E. angustifolia to respond positively to salt stress and enhances its salt tolerance by regulating the activities of some key proteins related to amino acid metabolism, lipid metabolism, and glutathione metabolism in root tissues. Conclusion: Aspartate aminotransferase, dehydratase-enolase-phosphatase 1 (DEP1), phospholipases D, diacylglycerol kinase, glycerol-3-phosphate O-acyltransferases, and gamma-glutamyl transpeptidases may play important roles in mitigating the detrimental effect of salt stress on mycorrhizal E. angustifolia . In conclusion, these findings provide new insights into the salt-stress tolerance mechanisms of AM E. angustifolia seedlings and also clarify the role of AM fungi in the molecular regulation network of E. angustifolia under salt stress.
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Grain sorghum has been a significant contributor to global food security since the prehistoric period and may contribute even more to the security of both food and energy in the future. Globally, precise management techniques are crucial for increasing grain sorghum productivity. In China, with diverse ecological types, variety introduction occasionally occurs across ecological zones. However, few information is available on the effect of ecological type on genotype performance and how plant spacing configuration influences grain yield in various ecological zones. Hence, a series of two-year field experiments were conducted in 2020 and 2021 in four ecological zones of China, from the northeast to the southwest. The experiments included six widely adapted sorghum varieties under six plant spacing configurations (two row spacing modes: equidistant row spacing (60 cm) mode and wide (80 cm)-narrow (40 cm) row spacing mode; three in-row plant spacings: 10 cm, 15 cm, and 20 cm). Our results indicated that ecological type, variety, and plant spacing configuration had a significant effect on sorghum yield. Ecological type contributed the highest proportion to the yield variance (49.8%), followed by variety (8.3%), while plant spacing configuration contributed 1.8%. Sorghum growth duration was highly influenced by the ecological type, accounting for 87.2% of its total variance, whereas plant height was mainly affected by genotype, which contributed 81.6% of the total variance. All test varieties, developed in the south or north, can reach maturity within 94-108 d, just before fall sowing in central China. Generally, sorghum growth duration becomes longer when a variety is introduced from south to north. A late-maturing variety, developed in the spring sowing and late-maturing regions, possibly could not reach maturity in the early-maturing region. The row spacing modes had no significant affect on sorghum yield, but the equal-row spacing mode consistently caused higher yields with only one exception; this might imply that equal-row spacing mode was more advantageous for boosting sorghum yield potential. In contrast, decreasing in-row plant spacing showed significant positive linear associations with sorghum grain yield in most cases. In addition, these results demonstrated that sorghum is a widely adapted crop and enables success in variety introduction across ecological zones.
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Forest degradation succession often leads to changes in forest ecosystem functioning. Exactly how the decomposition of leaf litter is affected in a disturbed forest remains unknown. Therefore, in our study, we selected a primary Korean pine forest (PK) and a secondary broad-leaved forest (SF) affected by clear-cutting degradation, both in Northeast China. The aim was to explore the response to changes in the leaf litter decomposition converting PK to SF. The mixed litters of PK and SF were decomposed in situ (1 year). The proportion of remaining litter mass, main chemistry, and soil biotic and abiotic factors were assessed during decomposition, and then, we made an in-depth analysis of the changes in the leaf litter decomposition. According to our results, leaf litter decomposition rate was significantly higher in the PK than that in the SF. Overall, the remaining percent mass of leaf litter's main chemical quality in SF was higher than in PK, indicating that leaf litter chemical turnover in PK was relatively faster. PK had a significantly higher amount of total phospholipid fatty acids (PLFAs) than SF during decomposition. Based on multivariate regression trees, the forest type influenced the soil habitat factors related to leaf litter decomposition more than decomposition time. Structural equation modeling revealed that litter N was strongly and positively affecting litter decomposition, and the changes in actinomycetes PLFA biomass played a more important role among all the functional groups. Selected soil abiotic factors were indirectly driving litter decomposition through coupling with actinomycetes. This study provides evidence for the complex interactions between leaf litter substrate and soil physical-chemical properties in affecting litter decomposition via soil microorganisms.
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In our previous paper, we analyzed "Deqi" in book Huangdi Neijing (The Yellow Emperor's Internal Classic) and Nanjing (Canon of Difficult Medical Problems) from "Zhishen"(Treating mentality) and Tiaoqi (Regulating qi). In the present paper, the authors discuss the connotations of "Deqi" and related events in the later ages of the abovementioned two classic books to the later stage of the Qing Dynasty when involves about 20 classical works as Zhenjiu Dacheng ( The Great Compendium of Acupuncture and Moxibustion), Zhenjing Zhinan (Guide to the classics of Acupuncture), Zhenjiu Daquan (A Complete Works of Acupuncture and Moxibustion) etc. from 1) close association between "Deqi" and patients' mental activity; 2) how to wait for arrival of qi if the needling does not induce "Deqi" for the time being; 3) how to identify "qi-arrival" and then, performing suitable manipulations; 4) Deqi and shallow- or deep-needling; 5) putting more emphasis on patients' feeling and reactions, rather than the practitioners perception beneath the needle which is described in book Huangdi Neijing; and 6) not withdrawing the acupuncture needles if qi does not arrive. Generally, in the later ages, the connotations of Deqi are enriched greatly.
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Terapia por Acupuntura/história , Livros/história , Qi/história , China , História Antiga , Humanos , Medicina na LiteraturaRESUMO
Background. Deqi sensation is a complex but an important component for acupuncture effect. In this study, we tried to observe the relationship between Deqi and skin temperature changes and whether there was some relativity between Deqi and needle stimulations on cold congealing and dysmenorrhea rat model. Thirty-two female Sprague Dawley (SD) rats were randomly divided into four groups (Saline Control Group, Model Group, Group A with strong stimulation, and Group B with small stimulation). Group A and Group B were performed with different stimulations. We found that, compared with saline control group, model group, and Group B, Group A showed that the skin temperature changes on right acupoint SP6 and SP10 increased significantly at 5 min-10 min interval. The skin temperature changes on left SP6 decreased at instant-5 min interval. The skin temperature changes on right SP10 decreased significantly at instant-5 min interval and 10 min-20 min interval. Thermogenic action along Spleen Meridian of Foot Greater Yin was manifested as simultaneous skin temperature increase on right SP6 and SP10 at 5 min-10 min interval after needling SP6, which was helpful to illustrate the relationship between the characteristic of Deqi and needle stimulations.
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Acupuncturists have always paid high attention to deqi (needling sensation) in clinical. However, relationship between deqi and curative effect has not been elucidated yet. In order to have a further understanding on effect of deqi on clinical effect, through study on literature associated with deqi since the establishment of People's Republic of China in 1949, it is held that various factors can influence therapeutic effect of acupuncture treatment, including whether needling sensation appears or not, the speed of deqi, the intensity of deqi, different types of sensations as well as propagated sensation along meridians (PSM). Joint launched multidisciplinary researches should be carried out to reveal the influence principle of deqi on acupuncture effect, and to lay foundations for the further studies on mechanism of deqi.
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Terapia por Acupuntura , Qi , Acupuntura , Pontos de Acupuntura , Humanos , Meridianos , Resultado do TratamentoRESUMO
In this study, we try to evaluate the effects of acupuncture stimulation with different amounts at Sanyinjiao (SP6) on uterine contraction and uterus microcirculation in rats with dysmenorrhea of cold coagulation syndrome and to explore whether there is direct relativity between "De qi" and needle stimulus intensity. Diestrus female rats were randomly divided into 4 groups, including saline control group, model control group, "A" stimulus group (with strong stimulus), and "B" stimulus group (with weak stimulus). We found that dysmenorrhea rats of the cold coagulation syndrome present a high intensity in uterine tension and high contraction of microvascular diameter. Acupuncture applied with two different stimuli could relieve the symptoms, but, compared with "B" stimulus, "A" stimulus leads to better outcomes on reducing uterine contraction and increasing diameter of uterine microvascular; moreover, hand manipulation during needling mediates the curative effect on the microvascular diameter. Our finding indicates that using thick needles and deep insertion with hand manipulation are more effective and achieve desired level of "De qi" in dysmenorrhea rats.
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Although it is difficult in fully clarifying its mechanisms and effects, Deqi still can be considered as an instant "sign" of acupuncture response of the patient and acupuncturist, which has a significant value in clinic and research. This paper aims to take a history trace to the development of Deqi theory, understand the connotation of Deqi based on Chinese medicine theory, and establish an evaluation methodology accordingly. We believe that Deqi is not only the needling sensation, but also the perception of changes of qi (') flowing of the patient elicited by needling on acupoints. The signs of Deqi include the patient's subjective perception (needling sensation), the objective physiological changes (common referred to the skin redness around the acupoints and the response of brain), and the acupuncturists' perception. Although Deqi is essential for attaining the effect, it may not be the necessary sign of the ideal efficacy. It is found that the characteristics of Deqi sensations, Deqi's intensity, time duration, and the propagation will all affect the efficacy. Thus, acupuncturists should pay attention to elicit and control Deqi state, which is also the key point in modern research on the therapeutic implications of Deqi.
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UNLABELLED: Deqi (needle sensation), is closely related to clinical therapeutic effect of acupuncture in the treatment of different clinical conditions. In the present paper, the authors summarized various components or concepts of Deqi mainly in the foreign studies collected from Pubmed database. At present, foreign researches about Deqi mainly include 1) the subjects' subjective qualitative and quantitative descriptions about Deqi, 2) correlation between acupuncture stimulation induced Deqi and clinical therapeutic effects, and 3) responses of different brain regions or the connectivity of brain network shown by functional magnetic resonance imaging (fMRI) during Deqi by needling different acupoints. According to the current commonly used 6 questionnaires, aching, tingling, numbness, heaviness, dull pain, throbbing, and deep pressure sensation are the most frequently seen components of Deqi of acupuncture stimulation. The factors influencing Deqi are psychological state, personal previous experience and cultural background. Regarding the degree of Deqi and clinical effects or outcomes, the conclusions are controversial. RESULTS: of fMRI analysis shows that in spite of extensive deactivation of the limbic system has been found in many studies, and the connectivity of the intrinsic brain functional networks is increased during acupuncture-induced Deqi, the significance of activation or deactivation of some brain regions remains unclear, and the related mechanisms need to be studied further. No matter activation or deactivation of different brain regions during Deqi, the most important issue is the relationship between Deqi and clinical therapeutic effects. Further studies are definitely needed.
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Terapia por Acupuntura , Qi , Encéfalo/diagnóstico por imagem , Encéfalo/fisiologia , Humanos , Imageamento por Ressonância Magnética , Radiografia , SensaçãoRESUMO
Deqi sensation is believed to be important in clinical efficacy according to TCM theory. The measuring method of Deqi sensation has significant implications for the result of research trials. This study makes an investigation on acupuncture-experienced patients and expert acupuncturists in China and aims to find out the patient's needling sensations and acupuncturist's sensations which can be acceptable as descriptors of Deqi sensation, so as to provide foundation for more systematic and sensitive quantitative evaluation method of Deqi sensation. Results of this survey indicated that the Deqi sensation noted by both patient and acupuncturist is equally important to the treatment efficacy. It is found that there are some differences between the patients' real-life experience and the acupuncturists' expectations on patients' Deqi sensation. The "dull pain," "aching," "sore," "numb," "distended," "heavy," "electric," "throbbing," "warmness," "coolness," "spreading," and "radiating" can be considered as the main manifestations of Deqi sensations. The acupuncturists believed that Deqi sensations were mainly "pulling," "tight," and "throbbing." We suggest developing a questionnaire measuring the Deqi sensations which includes both the sensations of the patient and acupuncturist, and this would be very important and necessary for a better understanding of the relationship between Deqi sensation and acupuncture effects in future studies.