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The genus Gnaphalium (Asteraceae) is distributed across most regions of the world. Some of them are not only used as a food source, but has also been used for the treatment of various diseases including pain, rheumatism, chronic pharyngitis and arthritis in China since ancient times. In this review, we would like to summarize previous research on genus Gnaphalium, including traditional uses, phytochemistry, bioactivities and quality control. The data presented here on genus Gnaphalium was generated based on various scientific research databases, including SciFinder, PubMed, ScienceDirect, Wiley library, Web of Science, and CNKI. Analysis of these findings showed that plants in genus Gnaphalium have a capital power in various therapeutic uses, including pulmonary protection, antimicrobial activity, antioxidant activity, anti-inflammatory and anti-complementary activities, and antidiabetic effect. Consistent with this, the chemicals from the plant extracts revealed its richness in various chemicals, including flavonoids, phenolic acids, terpenoids, sterols, and others. In spite of its wide applicable value worldwide, the quality control of genus Gnaphalium is still based on local standards. Thus, we argue that a national standard is required for genus Gnaphalium in China, to validate its bioactivity and future clinical trials rigorously.

The objective of this study is to explore the mechanism of action at the contact interface between gears and particles in spindle barrel finishing, using the Discrete Element Method (DEM) for simulation. The motion of the particles in the vicinity of the gear and the contact particles on the tooth surface is first described. Then the effects of gear embedment depth, gear and roller speed on relative particle motion velocity and tooth contact force are investigated. Finally, the simulation results are verified by experiments. The results show that the action of spindle barrel finishing on the gear tooth face is cyclical in nature. Contact force on the upper and lower tooth surfaces of the gear is not uniform, and the contact force on the upper tooth surface is 1.5 to 1.8 times that on the lower tooth surface. Increasing the gear embedment depth mainly affects the contact force between the particles and the tooth surface. A 75% increase in embedment depth leads to a 76% rise in tooth surface contact force. Similarly, increasing the gear and drum speed mainly affects the relative movement speed between particles and the tooth surface. A 150% increase in gear and drum speed results in a 148% increase in the relative movement speed of particles in contact with the tooth surface. Increasing the embedment depth of the gear can reduce the processing variability of the gear tooth surface along the axial direction. After increasing the embedment depth from 80 mm to 140 mm, the roughness of the upper and lower tooth surfaces along the axial direction decreases from 17% and 36% to 62% and 55%, respectively. However, the processing variability along the tooth profile direction does not change significantly by changing the speed and embedment depth.

Abstract Subnival glasshouse plants provide a text-book example of high-altitude adaptation with reproductive organs enclosed in specialized semi-translucent bracts, monocarpic reproduction and continuous survival under stress. Here, we present genomic, transcriptomic and metabolomic analyses for one such plant, the Noble rhubarb (Rheum nobile). Comparative genomic analyses show that an expanded number of genes and retained genes from two recent whole-genome duplication events are both relevant to subnival adaptation of this species. Most photosynthesis genes are downregulated within bracts compared to within leaves, and indeed bracts exhibit a sharp reduction in photosynthetic pigments, indicating that the bracts no longer perform photosynthesis. Contrastingly, genes related to flavonol synthesis are upregulated, providing enhanced defense against UV irradiation damage. Additionally, anatomically abnormal mesophyll combined with the downregulation of genes related to mesophyll differentiation in bracts illustrates the innovation and specification of the glass-like bracts. We further detect substantial accumulation of antifreeze proteins (e.g. AFPs, LEAs) and various metabolites (e.g. Proline, Protective sugars, procyanidins) in over-wintering roots. These findings provide new insights into subnival adaptation and the evolution of glasshouse alpine plants.

Resource-transfer connections among bacteria, fungi, and microbivorous nematodes play an integral role in the decomposition pathways of soil micro-foodweb. Exploring the response of soil bacteria, fungi, and nematodes to grassland degradation is crucial for predicting the direction and trajectory of degradation. However, the knowledge about soil bacteria-fungi-nematode interactions in ecosystem function and nutrient cycling must be highlighted properly, especially in the Qinghai-Tibet Plateau. Therefore, this study focuses on four alpine meadows at different degenerate gradients (non-degraded grassland, slightly degraded grassland, moderately degraded grassland, and severely degraded grassland) on the Qinghai-Tibet Plateau. The investigation of soil bacteria-fungi-nematode interactions uses high-throughput sequencing and correlation network analysis. The results demonstrate significant variations in the composition of soil nematode genera, and bacterial and fungal phyla, across different degradation stages. Moreover, the diversity of nematodes, bacteria and fungi declined as grassland degradation worsened. Principal component analysis (PCA) revealed that alpine meadows with varying degradation gradients significantly influenced the community structure of bacteria, fungi, and nematodes. Mantel analysis indicated that soil water content (P

Characterizing the photosynthetic capacity of grassland is essential for exploring the carbon cycle under climate change. However, little research has been conducted to date on photosynthetic accumulation per leaf area (NPP/LAI) in spatial and temporal terms, especially on how they respond to climate change. Hence, this study analyzed the spatiotemporal patterns of NPP/LAI based on MODIS products, explored the partial correlation of NPP/LAI with climatic factors for the annual and growing season, and quantified the time lag effects of climatic factors on NPP/LAI for seasonal in alpine grassland ecosystems at the source of the Yellow River Basin. The results indicated that NPP, LAI, and NPP/LAI of growing season increased in the study area between 2000 and 2021 whiles annual NPP/LAI decreased. At the same time, the spatial distribution of the multiannual mean values of precipitation and temperature approximately reversed and gradually increased on the time scale. The annual NPP/LAI for each grassland type gradually decreased between 2000 and 2021, with the mean of the growing season in NPP/LAI ranging from montane meadows (12.595 g C·m–2) > swamps (11.859 g C·m–2) > alpine meadows (11.718 g C·m–2) > temperate steppes (10.994 g C·m–2). It is also interesting to note that the mean values for montane meadows and alpine meadows were slightly higher in summer than in autumn, while the opposite was true for temperate steppes and swamps. Regarding the effects of climate, the response of NPP/LAI to temperature and precipitation varied among grassland types on an annual and seasonal basis. In addition, the seasonal lag effect of climate factors showed that the overall degree of effect of winter temperature on NPP/LAI in subsequent seasons increased as the seasonal interval progressed, while the opposite was true for precipitation, meaning that temperature affected NPP/LAI over a longer period compared to precipitation. Moreover, the seasonal time lag effects of climatic factors in different grassland types had varying degrees of influence on NPP/LAI, with the NPP/LAI of montane meadows and alpine grasslands being more closely related to climatic factors than those of temperate steppes and swamps. This study provides a basis for effective monitoring of photosynthetic capacity dynamics and helps develop policies to cope with climate change in alpine grasslands.

Understanding the large-scale distribution patterns of invasive alien plants can not only identify potential high-risk areas but also provide key information for developing management strategies to mitigate plant invasion. Here, for the first time, we mapped the spatial distribution of 239 invasive plant species in China at much finer spatial resolution (50 × 50 km2). We also developed a new metric, invasion level score (ILS), which represent both invasive plant richness and the number of high-risk invasive plants per grid cell, to identify hotspots of plant invasion. Finally, we compared differences in invasive plant richness and ILS between protected and non-protected areas. We found that the counties and grids in the southern and eastern coastal regions of China have the highest species richness and ILS. The richness and ILS decrease from southeast to northwest China. Meanwhile, the high-risk areas for invasion level are concentrated in and around provincial capitals in the southern, eastern, and northern China. We also found that protected areas in China have significantly more invasive plants and higher ILS than non-protected areas. The grid cells with high invasive plant richness and ILS are mostly distributed in regions with high population density, likely due to urban effect. The need to direct management attention towards the southern and eastern China and particularly within protected areas was suggested by our findings. Our study provides a comprehensive assessment of the distribution and risk of invasive plants in China, as well as a scientific foundation for further advancing the prevention and management of invasive plants.

Shift work may adversely affect individuals’ health, thus, the current study aimed to investigate the association between shift work and health outcomes in the general population. A total of 41,061 participants were included in this online cross-sectional survey, among which 9612 (23.4%) individuals engaged in shift work and 31,449 (76.6%) individuals engaged in non-shift work. Multiple logistic regression analyses were conducted to explore the association between shift work and health outcomes (psychiatric disorders, mental health symptoms, and physical disorders). In addition, associations between the duration (≤1 year, 1–3 years, 3–5 years, 5–10 years, ≥10 years) and frequency of shift work (p < 0.001), mental health symptoms (OR = 1.76, 95% CI = 1.68–1.85, p < 0.001), and physical disorders (OR = 1.48, 95% CI = 1.39–1.57, p < 0.001). In addition, inverted U-shaped associations were observed between the duration of shift work and health outcomes. These results indicated that shift work was closely related to potential links with poor health outcomes. The findings highlighted the importance of paying attention to the health conditions of shift workers and the necessity of implementing comprehensive protective measures for shift workers to reduce the impact of shift work.

Coronavirus disease 2019 (COVID-19) has rapidly spread worldwide. Traditional Chinese Medicine (TCM) was considered important by Chinese health authorities in the fight against COVID-19. This review systematically analyzed and evaluated the safety and efficacy of TCM combined with Western Medicine (WM) for the treatment of COVID-19. We sought to provide summary evidence for clinicians when using TCM. We searched for studies in PubMed, Web of Science, Embase, Medline, the Cochrane Library, China National Knowledge Infrastructure, and Wanfang Data from database inception to June 1, 2021. Overall, 31 studies (14,579 participants) were involved in the final systematic review, including 15 randomized controlled trials and 16 observational studies. TCM combined with WM showed main outcomes of a higher clinical efficacy rate (odds ratio [OR] =2.48, 95% confidence interval [CI] =1.90–3.24, I2 = 4%) and lower case fatality rate (OR = 0.31, 95% CI = 0.19–0.49, I2 = 80%) compared with WM treatment alone. No significant overall adverse events were found between TCM plus WM group and WM group (OR = 1.43, 95% CI = 0.63–2.23, I2 = 75%). Some larger randomized control trials would assist in defining the effect of TCM combined with WM on the treatment of COVID-19 complications such as cardiac injury. TCM combined with WM may be safe and effective for the treatment of COVID-19.

Abstract Isatis indigotica Fort. (Chinese woad) is a species with an ancient and well-documented history as an indigo dye and medicinal plant. It is often confused with Isatis tinctoria L. (European woad), a medicinal plant in Europe. Here, the differences between I. indigotica and I. tinctoria are systematically described. The usage development history, clinical applications and pharmacological activities, and chemical components of I. indigotica are also summarized. Lignans, indole alkaloids, and their corresponding derivatives have been identified as the major active ingredients of I. indigotica and are associated with anti-viral, anti-inflammatory, anti-cancer, and other health-promoting activities. Notable progress has been made in understanding the biosynthetic pathway and regulation mechanism of lignans and indole alkaloids in I. indigotica, the results from which should facilitate the process of targeted metabolic engineering or synthetic biology. Moreover, multiple biotechnology methods such as polyploid breeding and genetic engineering have been used with I. indigotica to result in, for example, greater yields, higher levels of bioactive component accumulation, and enhanced stress tolerance to salt, drought, and insects. Some issues require additional analyses, and suggestions for future research on I. indigotica are also discussed.

Cofactors are crucial for the biosynthesis of natural compounds, and cofactor engineering is a useful strategy for enzyme optimization due to its potential to enhance enzyme efficiency. Secoisolariciresinol dehydrogenase (SIRD) was reported to convert secoisolariciresinol into matairesinol in an NAD+-dependent reaction. Here, a SIRD designated as IiSIRD2 identified from Isatis indigotica was found to utilize NADP+ as the cofactor. To explore the structural basis for this unique cofactor preference, model-based structural analysis was carried out, and it was postulated that a variation at the GXGGXG glycine-rich motif of IiSIRD2 alters its cofactor preference. This study paves way for future investigations on SIRD cofactor specificity and cofactor engineering to improve SIRD’s catalytic efficiency.

Soil water–salt variations at different spatiotemporal scales and their influencing factors are essential for regional vegetation restoration and management. However, the study of soil water–salt of the alpine area has not taken into account the effect of topography, especially in the long time series. This work investigated the spatiotemporal characteristics of water and salt in topsoil from 2000 to 2021, using regression analysis of on-site data and MODIS products. The heterogeneity and trends of soil water–salt in various topographic factors (elevation, slope, and aspect) were also explored by selecting the maximum area of each gradient factor as the constrained condition with the help of ArcGIS software. The results revealed that 1) the models applied to SWC (soil water content), EC (electrical conductivity), and pH utilizing vegetation indices were suitable for estimating the spatiotemporal variations of soil water–salt. 2) Soil water–salt exhibited local aggregation characteristics with an upward trend in SWC and a downward trend in EC and pH as a whole. 3) Mean annual SWC peaked at 3,400–3,800 m with increasing elevation and trended downward with increasing slope. The aspect of mean annual SWC was characterized by lower values in the southwest and south and higher values in the north, while the results for pH were reversed. The slope had a greater influence on EC than on SWC and pH. 4) The trend change of SWC, EC, and pH fluctuated at 2000–2,400 m and >4,600 m with a clear inflection point at 4°–8° and 20°–24°, respectively. In summary, the effects of topography responded to the distribution and trends of soil water–salt. The results have significant implications for implementing ecological improvements in regions with complicated topography and can serve as a reference for formulating future ecological policies.

B-box (BBX) proteins are one of the zinc-finger transcription factor that plays a critical role in plant development, growth, and multiple stress responses. Although BBX genes have been reported in many model organisms, no comprehensive study has yet been conducted on the BBX genes in Melilotus albus, and the biological functions of this family remain unknown. In this study, a total of 20 BBX (MaBBX) genes were identified in M. albus and were phylogenetically divided into five clades. BBX members within the same clade showed similar conserved domain, suggesting similarity of potential biological function. Analysis of MaBBX conserved motifs showed that every subfamily contained two common motifs. Distribution mapping shows that BBX proteins are nonrandomly localized in eight chromosomes. The synteny showed that most homologous gene pairs of the MaBBX gene family were amplified by segmental replication, which meant segmental replication was the main way for the MaBBX gene family to evolve. Additionally, the cis-element analysis predicted light-responsive, various hormone and stress-related elements in the promoter regions of MaBBXs. Furthermore, the expression levels of all 20 MaBBX genes were detected by qRT-PCR under salt, cold, and dark stresses in M. albus. Moreover, it was observed that 16 genes had higher expression levels after 3 h of salt treatment, 10 genes were significantly upregulated after 3 h of cold treatment, and all genes were up regulated after 3 h of dark treatment, and then appeared to decline. In addition, it was also noticed that MaBBX13 may be an important candidate for improving tolerance to abiotic stress. The prediction of protein tertiary structure showed that the tertiary structures of members of the same subfamily of MaBBX proteins were highly similar. The hypothesis exhibited that most of the MaBBX proteins were predicted to be localized to the nucleus and cytoplasm and was validated by transient expression assays of MaBBX15 in tobacco leaf epidermal cells. This study provides useful information for further investigating and researching the regulatory mechanisms of BBX family genes in response to abiotic stresses in M. albus.

Obesity and its associated complications are highly related to a current public health crisis around the world. A growing body of evidence has indicated that G-protein coupled bile acid (BA) receptor TGR5 (also known as Gpbar-1) is a potential drug target to treat obesity and associated metabolic disorders. We have identified notoginsenoside Ft1 (Ft1) from Panax notoginseng as an agonist of TGR5 in vitro. However, the pharmacological effects of Ft1 on diet-induced obese (DIO) mice and the underlying mechanisms are still elusive. Here we show that Ft1 (100 mg/100 diet) increased adipose lipolysis, promoted fat browning in inguinal adipose tissue and induced glucagon-like peptide-1 (GLP-1) secretion in the ileum of wild type but not Tgr5−/− obese mice. In addition, Ft1 elevated serum free and taurine-conjugated bile acids (BAs) by antagonizing Fxr transcriptional activities in the ileum to activate Tgr5 in the adipose tissues. The metabolic benefits of Ft1 were abolished in Cyp27a1−/− mice which have much lower BA levels. These results identify Ft1 as a single compound with opposite activities on two key BA receptors to alleviate high fat diet-induced obesity and insulin resistance in mice.

Pinoresinol–lariciresinol reductases (PLRs) are enzymes involved in the lignan biosynthesis. Here, crystal structures of three PLRs in the apo, substrate-bound and product-bound states, and accompanying mutagenesis provide insight into PLRs catalytic mechanism and suggest a strategy for PLR engineering.

Panax species have gained numerous attentions because of their various biological effects on cardiovascular, kidney, reproductive diseases known for a long time. Recently, advanced analytical methods including thin layer chromatography, high-performance thin layer chromatography, gas chromatography, high-performance liquid chromatography, ultra-high performance liquid chromatography with tandem ultraviolet, diode array detector, evaporative light scattering detector, and mass detector, two-dimensional high-performance liquid chromatography, high speed counter-current chromatography, high speed centrifugal partition chromatography, micellar electrokinetic chromatography, high-performance anion-exchange chromatography, ambient ionization mass spectrometry, molecularly imprinted polymer, enzyme immunoassay, 1H-NMR, and infrared spectroscopy have been used to identify and evaluate chemical constituents in Panax species. Moreover, Soxhlet extraction, heat reflux extraction, ultrasonic extraction, solid phase extraction, microwave-assisted extraction, pressurized liquid extraction, enzyme-assisted extraction, acceleration solvent extraction, matrix solid phase dispersion extraction, and pulsed electric field are discussed. In this review, a total of 219 articles published from 1980 to 2018 are investigated. Panax species including P. notoginseng, P. quinquefolius, sand P. ginseng in the raw and processed forms from different parts, geographical origins, and growing times are studied. Furthermore, the potential biomarkers are screened through the previous articles. It is expected that the review can provide a fundamental for further studies.

Yield potential, pharmaceutical compounds production and stress tolerance capacity are 3 classes of traits that determine the quality of medicinal plants. The autotetraploid Isatis indigotica has greater yield, higher bioactive lignan accumulation and enhanced stress tolerance compared with its diploid progenitor. Here we show that the transcription factor IiWRKY34, with higher expression levels in tetraploid than in diploid I. indigotica, has large pleiotropic effects on an array of traits, including biomass growth rates, lignan biosynthesis, as well as salt and drought stress tolerance. Integrated analysis of transcriptome and metabolome profiling demonstrated that IiWRKY34 expression had far-reaching consequences on both primary and secondary metabolism, reprograming carbon flux towards phenylpropanoids, such as lignans and flavonoids. Transcript–metabolite correlation analysis was applied to construct the regulatory network of IiWRKY34 for lignan biosynthesis. One candidate target Ii4CL3, a key rate-limiting enzyme of lignan biosynthesis as indicated in our previous study, has been demonstrated to indeed be activated by IiWRKY34. Collectively, the results indicate that the differentially expressed IiWRKY34 has contributed significantly to the polyploidy vigor of I. indigotica, and manipulation of this gene will facilitate comprehensive improvements of I. indigotica herb.

Backgroud: Sleep deprivation (SD) impairs learning and memory by inhibiting hippocampal functioning at molecular and cellular levels. Abnormal autophagy and apoptosis are closely associated with neurodegeneration in the central nervous system. This study is aimed to explore the alleviative effect and the underlying molecular mechanism of stem–leaf saponins of Panax notoginseng (SLSP) on the abnormal neuronal autophagy and apoptosis in hippocampus of mice with impaired learning and memory induced by SD. Methods: Mouse spatial learning and memory were assessed by Morris water maze test. Neuronal morphological changes were observed by Nissl staining. Autophagosome formation was examined by transmission electron microscopy, immunofluorescent staining, acridine orange staining, and transient transfection of the tf-LC3 plasmid. Apoptotic event was analyzed by flow cytometry after PI/annexin V staining. The expression or activation of autophagy and apoptosis-related proteins were detected by Western blotting assay. Results: SLSP was shown to improve the spatial learning and memory of mice after SD for 48 h, accomanied with restrained excessive autophage and apoptosis, whereas enhanced activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in hippocampal neurons. Meanwhile, it improved the aberrant autophagy and apoptosis induced by rapamycin and re-activated phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signaling transduction in HT-22 cells, a hippocampal neuronal cell line. Conclusion: SLSP could alleviate cognitive impairment induced by SD, which was achieved probably through suppressing the abnormal autophagy and apoptosis of hippocampal neurons. The findings may contribute to the clinical application of SLSP in the prevention or therapy of neurological disorders associated with SD. Keywords: Apoptosis, Autophagy, Learning and memory, Panax notoginseng, Sleep deprivation, Stem-leaf saponins

Diabetes is a common and complex disease that can exacerbate the complications related to cardiovascular disease, and this is especially true for type 2 diabetes mellitus (T2DM). In addition to the standard pharmacological therapies, T2DM has also been treated with nonconventional regimens such as traditional Chinese medicine (TCM), e.g., herbal medicines and TCM prescriptions, although the mechanisms underlying the therapeutic benefits remain unclear. In this regard, many studies have used metabolomics technology to elucidate the basis for the efficacy of TCM for T2DM. Metabolomics has recently attracted much attention with regard to drug discovery and pharmacologically relevant natural products. In this review, we summarize the application of metabolomics to the assessment of TCM efficacy for treating T2DM. Increasing evidence suggests that the metabolic profile of an individual patient may reflect a specific type of T2DM syndrome, which may provide a new perspective for disease diagnosis. In addition, TCM has proved effective for countering the metabolic disorders related to T2DM, and this may constitute the basis for TCM efficacy. Therefore, further determining how TCM contributes to the reversal of metabolic disorders, such as using network pharmacology or by assessing the contribution of host–gut microbiota interactions, will also provide researchers with new potential targets for pharmacologic-based therapies.

Sleep deprivation (SD) may lead to serious myocardial injury in cardiovascular diseases. Saponins extracted from the roots of Panax notoginseng, a traditional Chinese medicine beneficial to blood circulation and hemostasis, are the main bioactive components exerting cardiovascular protection in the treatment of heart disorders, such as arrhythmia, ischemia and reperfusion injury, and cardiac hypertrophy. This study aimed to explore the protective effect of stem-leaf saponins from Panax notoginseng (SLSP) on myocardial injury in SD mice. SD was induced by a modified multi-platform method. Cardiac morphological changes were assessed by hematoxylin and eosin (H&E) staining. Heart rate and ejection fraction were detected by specific instruments. Serum levels of atrial natriuretic peptide (ANP) and lactate dehydrogenase (LDH) were measured with biochemical kits. Transmission electron microscopy (TEM), immunofluorescent, and Western blotting analysis were used to observe the process and pathway of autophagy and apoptosis in heart tissue of SD mice. In vitro, rat H9c2 cells pretreated with rapamycin and the effect of SLSP were explored by acridine orange staining, transient transfection, flow cytometry, and Western blotting analysis. SLSP prevented myocardial injury, such as morphological damage, accumulation of autophagosomes in heart tissue, abnormal high heart rate, serum ANP, and serum LDH induced by SD. In addition, it reversed the expressions of proteins involved in the autophagy and apoptosis and activated PI3K/Akt/mTOR signaling pathway that is disturbed by SD. On H9c2 cells induced by rapamycin, SLSP could markedly resume the abnormal autophagy and apoptosis. Collectively, SLSP attenuated excessive autophagy and apoptosis in myocardial cells in heart tissue induced by SD, which might be acted through activating PI3K/Akt/mTOR signaling pathway.

The plant-specific SHI-related sequence (SRS) family of transcription factors plays a vital role in growth regulation, plant development, phytohormone biosynthesis, and stress response. However, the genome-wide identification and role in the abiotic stress-related functions of the SRS gene family were not reported in white sweet clover (Melilotus albus). In this study, nine M. albus SRS genes (named MaSRS01-MaSRS09) were identified via a genome-wide search method. All nine genes were located on six out of eight chromosomes in the genome of M. albus and duplication analysis indicated eight segmentally duplicated genes in the MaSRS family. These MaSRS genes were classified into six groups based on their phylogenetic relationships. The gene structure and motif composition results indicated that MaSRS members in the same group contained analogous intron/exon and motif organizations. Further, promoter region analysis of MaSRS genes uncovered various growth, development, and stress-responsive cis-acting elements. Protein interaction networks showed that each gene has both functions of interacting with other genes and members within the family. Moreover, real-time quantitative PCR was also performed to verify the expression patterns of nine MaSRS genes in the leaves of M. albus. The results showed that nine MaSRSs were up- and down-regulated at different time points after various stress treatments, such as salinity, low-temperature, salicylic acid (SA), and methyl jasmonate (MeJA). This is the first systematic study of the M. albus SRS gene family, and it can serve as a strong foundation for further elucidation of the stress response and physiological improvement of the growth functions in M. albus.

Abstract Background Ruili is a border city in southwest China along the heroin trafficking route. In recent decades, the city has witnessed increased in HIV transmission. The current study aims to explore the spatiotemporal trends in HIV prevalence identify and map the spatial variation and clustering of factors associated with HIV transmission through drug use and heterosexual contact transmissions at the village level from 1989 through 2016. Methods Geographic information system-based spatiotemporal analyses, including global and local spatial autocorrelation analyses and space-time scanning statistics, were applied to detect the location and extent of HIV/AIDS high-risk areas. Results Drug use and heterosexual contact were identified as the major transmission routes causing infection in Ruili. Results of global spatial analysis showed significant clustering throughout the city caused by transmission via drug use in the early phase of the epidemic and transmission via heterosexual contact in the late phase of the epidemic during the study period. Hotspots of transmission from drug use were randomly distributed throughout the city. However, the hotspots of transmission by heterosexual contact were located in the central area only around the Jiegao China-Myanmar land port. Space-time scanning showed that transmission from drug use clustered in the southwest area between 1989 and 1990, while transmission by heterosexual contact clustered in the central area between 2004 and 2014. Conclusions Heterosexual contact has become the dominant mode of transmission. Interventions should focus on highly clustered area where is around the Jiegao land port.

Background: Ginsenosides are known as the principal pharmacological active constituents in Panax medicinal plants such as Asian ginseng, American ginseng, and Notoginseng. Some ginsenosides, especially the 20(R) isomers, are found in trace amounts in natural sources and are difficult to chemically synthesize. The present study provides an approach to produce such trace ginsenosides applying biotransformation through Escherichia coli modified with relevant genes. Methods: Seven uridine diphosphate glycosyltransferase (UGT) genes originating from Panax notoginseng, Medicago sativa, and Bacillus subtilis were synthesized or cloned and constructed into pETM6, an ePathBrick vector, which were then introduced into E. coli BL21star (DE3) separately. 20(R)-Protopanaxadiol (PPD), 20(R)-protopanaxatriol (PPT), and 20(R)-type ginsenosides were used as substrates for biotransformation with recombinant E. coli modified with those UGT genes. Results: E. coli engineered with GT95syn selectively transfers a glucose moiety to the C20 hydroxyl of 20(R)-PPD and 20(R)-PPT to produce 20(R)-CK and 20(R)-F1, respectively. GTK1- and GTC1-modified E. coli glycosylated the C3OH of 20(R)-PPD to form 20(R)-Rh2. Moreover, E. coli containing p2GT95synK1, a recreated two-step glycosylation pathway via the ePathBrich, implemented the successive glycosylation at C20OH and C3OH of 20(R)-PPD and yielded 20(R)-F2 in the biotransformation broth. Conclusion: This study demonstrates that rare 20(R)-ginsenosides can be produced through E. coli engineered with UTG genes. Keywords: biosynthesis, 20(R)-ginsenosides, ginsenoside, UDP-glycosyltransferase

The LIM (Lin-11, Isl-1 and Mec-3 domains) family is a key transcription factor widely distributed in animals and plants. The LIM proteins in plants are involved in the regulation of a variety of biological processes, including cytoskeletal organization, the development of secondary cell walls, and cell differentiation. It has been identified and analyzed in many species. However, the systematic identification and analysis of the LIM genes family have not yet been reported in alfalfa (Medicago sativa L.). Based on the genome-wide data of alfalfa, a total of 21 LIM genes were identified and named MsLIM01-MsLIM21. Comprehensive analysis of the chromosome location, physicochemical properties of the protein, evolutionary relationship, conserved motifs, and responses to abiotic stresses of the LIM gene family in alfalfa using bioinformatics methods. The results showed that these MsLIM genes were distributed unequally on 21 of the 32 chromosomes in alfalfa. Gene duplication analysis showed that segmental duplications were the major contributors to the expansion of the alfalfa LIM family. Based on phylogenetic analyses, the LIM gene family of alfalfa can be divided into four subfamilies: αLIM subfamily, βLIM subfamily, γLIM subfamily, and δLIM subfamily, and approximately all the LIM genes within the same subfamily shared similar gene structure. The 21 MsLIM genes of alfalfa contain 10 Motifs, of which Motif1 and Motif3 are the conserved motifs shared by these genes. Furthermore, the analysis of cis-regulatory elements indicated that regulatory elements related to transcription, cell cycle, development, hormone, and stress response are abundant in the promoter sequence of MsLIM genes. Real-time quantitative PCR demonstrated that MsLIM gene expression is induced by low temperature and salt. The present study serves as a basic foundation for future functional studies on the alfalfa LIM family.

Alpha-glucosidase inhibitors currently form an important basis for developing novel drugs for diabetes treatment. In our preliminary tests, the ethyl acetate fraction of Phlomis tuberosa extracts showed significant α-glucosidase inhibitory activity (IC₅₀ = 100 μg/mL). In the present study, a combined method using Sepbox chromatography and thin-layer chromatography (TLC) bioautography was developed to probe α-glucosidase inhibitors further. The ethyl acetate fraction of P. tuberosa extracts was separated into 150 individual subfractions within 20 h using Sepbox chromatography. Then, under the guidance of TLC bioautography, 20 compounds were successfully isolated from these fractions, including four new diterpenoids [14-hydroxyabieta-8,11,13-triene-11-carbaldehyde-18-oic-12-carboxy-13-(1-hydroxy-1-methylethyl)-lactone (1), 14-hydroxyabieta-8,11,13-triene-17-oic-12-carboxy-13-(1-hydroxy-1-methylethyl)-lactone (2), 14,16-dihydroxyabieta-8,11,13-triene-15,17-dioic acid (3), and phlomisol (15,16-eposy-8,13(16),14-labdatrien-19-ol) (4)], and 16 known compounds. Activity estimation indicated that 15 compounds showed more potent α-glucosidase inhibitory effects (with IC50 values in the range 0.067-1.203 mM) than the positive control, acarbose (IC50 = 3.72 ± 0.113 mM). This is the first report of separation of α-glucosidase inhibitors from P. tuberosa.

Cofactors are crucial for the biosynthesis of natural compounds, and cofactor engineering is a useful strategy for enzyme optimization due to its potential to enhance enzyme efficiency. Secoisolariciresinol dehydrogenase (SIRD) was reported to convert secoisolariciresinol into matairesinol in an NAD+-dependent reaction. Here, a SIRD designated as IiSIRD2 identified from Isatis indigotica was found to utilize NADP+ as the cofactor. To explore the structural basis for this unique cofactor preference, model-based structural analysis was carried out, and it was postulated that a variation at the GXGGXG glycine-rich motif of IiSIRD2 alters its cofactor preference. This study paves way for future investigations on SIRD cofactor specificity and cofactor engineering to improve SIRD's catalytic efficiency. [ABSTRACT FROM AUTHOR]