Ascochyta, Botryosphaeria, Saccharomyces, Nectriaceae_NA, Trametes, and Valsaceae_NA. However, the microbiome also included bacteria beneficial to plant growth and the production of polysaccharides—namely, 1174-901-12, Catenibacterium, Cutibacterium, Friedmanniella, Methylobacterium-Methylorubrum, Pseudomonas, Rhodobacter, and Sphingomonas. Furthermore, we confirmed that gummosis induced a more complex structure in the bark microbiome network. We conclude that the findings of this study provide a valuable aid in profiling the overall peach tree microbial ecosystem, which can be utilized to develop precise biomarkers for the early diagnosis of gummosis.Peach gummosis disease has been identified as a serious challenge in Korean agriculture and has developed to become a major cause of agricultural productivity losses. However, treatments for gummosis have not been systemically established and studies of the microbiome closely related to this plant disease are lacking. Therefore, we analyzed the bacterial and fungal communities in the bark and rhizosphere soil of healthy peach trees and those with gummosis. Through high-throughput sequencing, we obtained unprecedented insights into the bacterial and fungal dynamics of each group, including their diversity and taxonomic classification, as well as network analyses. We found that the presence of gummosis drives a significantly higher alpha diversity in the bark bacterial community. Peach gummosis bark mycobiomes included greater numbers of opportunistic pathogens such as  Peaches are the third most cultivated fruit crop in South Korea, making the country one of the leading producers of peaches in the world. The area under peach tree cultivation is approximately 20,000 ha, accounting for 13.2% of the total fruit tree cultivation area in the country. Gummosis, a disease of fruit trees, is characterized by a dark brown resin secreted from the stems or branches of the tree and occurs periodically throughout the cultivation cycle , and conBotryosphaeria species—including B. dothidea, B. obtusa, and B. rhodian—are commonly detected in peaches [Pseudomonas syringae—which causes bacterial gummosis—has been reported [Although factors such as environmental stress and physical or chemical injuries may partly determine the occurrence or severity of gummosis , infecti peaches ,7,8,9,10reported ,12. Studreported ,14,15,16Peach trees are vulnerable to latent pathogenic infections year-round. There are as of yet no effective or approved fungicides for the control of peach gummosis, and although pesticides are used to prevent fruit and leaf diseases, such applications can prove expensive ,18,19. Tp < 0.0001 for Observed and Shannon, and p < 0.001 for InvSimpson, respectively; p > 0.05; p < 0.001; p < 0.001; p = 0.001 and p = 0.004, respectively, for bark and soil analyses; p = 0.035; First, we evaluated the alpha diversity of the microbial community with and without the presence of gummosis. In the case of the Observed, Shannon, and InvSimpson indices, the bacterial diversity of the peach gummosis bark (PGB) was far higher than that of the healthy peach bark  in the bark, and three genera  in the soil  effect size (LEfSe) analysis. The LEfSe results, showing the optimal features to account for group differences by LDA score, indicated genera with LDA scores of 3.5 or higher that were representative of each group. In multiple comparisons, including HPB–PGB and HPS–PGS, numerous taxa were identified to discriminate between individual groups. Specifically, the microbial community in the HPB comprised 16 genera (6 bacteria and 10 fungi), whereas PGB comprised 33 genera  consisted of 10, 20, 41, and 41 nodes and 3, 55, 137, and 168 edges, respectively . NetworkMicrobiomes are widely perceived as a requisite portion of the crop ecosystem and are strongly associated with plant growth and disease resistance ,20,21,22Microvirga, Nitrospira, and Sphingomonas, have been reported in healthy soils or as being involved in nitrification. There are numerous reasons why this phenomenon might exist. The rhizosphere may only be slightly impacted by the gummosis disease that emerges in the bark, or it may even be a microbial defense mechanism against plant stress. Numerous previous studies have demonstrated that plants can employ a “cry for help” strategy to enhance their ability to resist stress from illnesses, and this strategy is consistent with our findings [To date, the diversity of the rhizosphere microflora in relation to plant diseases has been extensively reported through studies on plant–microbial interactions ,36,37,38findings ,40.The microbial community under gummosis in the bark differed noticeably from that in the soil. First, a comparison of HPB and PGB showed only a significant difference in alpha diversity, indicating which community should be focused on—the rhizosphere or endophytic—to understand microflora changes following gummosis. Alpha diversity indices have been used as indicators of host plant health in prior studies, although this assertion is controversial. According to Yuan et al. , the div1174-901-12 , Sphingomonas, Methylobacterium-Methylorubrum, and Pseudomonas are commonly increased in plants infected with Pseudomonas syringae pv. actinidiae, suggesting the potential antagonistic properties of these bacteria against phytopathogenic fungi. This pilot study supports our hypothesis that host plants provide available resources to stimulate the development of beneficial endophytes in response to gummosis. In addition, the considerable surge in Sphingomonas following gummosis, regardless of the rhizosphere and endosphere, emphasizes the need for further research on the role of microbes in the environment.Significant changes in taxonomic classification between HPB and PGB clarify the structural changes in bacterial communities. We discovered an intriguing characteristic of microbes that was drastically increased in PGB. Contrary to our assumptions, the microbial community in the bark of trees with gum disease was verified to contain far more microorganisms conducive to plant growth than pathogenic microbes. In addition, the majority of these microorganisms produce polysaccharides ,44,45,46Pseudomonas, Methylobacterium-Methylorubrum, Sphingomonas, 1174-901-12, and Catenibacterium) out of 27 bacterial genera representing PGB selected with high LDA scores through LEfSe showed remarkable differences even in comparison to other 22 taxa, strengthening their potential as biomarker candidates. Among the fungi, Alternaria is the only taxon that differs statistically in both LEfSe and relative abundance comparisons between HPB and PGB. Surprisingly, we found that significantly increased fungal groups, including Alternaria, were predominantly composed of opportunistic plant pathogens, such as Ascochyta, Botryosphaeria, Saccharomyces, Nectriaceae_NA, Trametes, and Valsaceae_NA . DNA extraction from pretreated bark pieces and 0.25 g of soil was performed using a DNeasy PowerSoil Kit , following the manufacturer’s instruction. The validity of the extracted DNA was confirmed through electrophoresis, and DNA was quantified using a Qubit fluorometer 2.0 .For the sequencing library preparation, V4–V5—the hypervariable region of the 16S rRNA gene—was amplified using a 515 F (5′-barcode-CGCTCTTCCGATCTGTGNCAGCMGCCGCGGTRA-3′) forward primer and a 907 R (5′-barcode-GTGCTCTTCCGATCCGYCWATTYHTTTRAGTTT-3′) reverse primer; the eukaryotic ITS2 region was amplified with the primer pairs ITS86F/ITS4R . The specific PCR conditions used have been described in a previous study . We thenRaw sequencing data were processed using the Quantitative Insights into Microbial Ecology2 (QIIME2) pipeline (version 2021.4) . In the To conduct the statistical analysis of the microbial community structure between HPB and PGB and HPS and PGS, multiple packages in R and GraphPad Prism 8.0.2 for Windows  were employed with respect to both bacterial and fungal communities in the bark and soil. For a better understanding, we monitored various alpha diversity indices—observed, Shannon, and InvSimpson—using the “phyloseq” R package . AnalyseFurthermore, we assessed the taxonomic distribution between the HP and PG groups at the phylum, family, and genus levels. Considering visibility, microbial classifications at the phylum and family levels were described in donut plots, and genera were separately displayed as a heatmap using the “pheatmap” R package. In the heatmap, genera with lower than 10% prevalence and 1% relative abundance were filtered out. To identify the genera that showed statistically significant differences between HP and PG in soil and bark, the Mann–Whitney test was implemented, and those groups with higher taxa abundances were marked with an asterisk.https://huttenhower.sph.harvard.edu/galaxy/ (accessed on 21 August 2022)). Only candidate taxa with LDA scores of 3.5 or greater were visualized.LEfSe analysis, an estimation of specific taxa in which attributes differ significantly depending on the existence of gummosis, was performed using the Galaxy implementation (p-value of less than 0.05 were visualized.Finally, we performed a network analysis to examine how genera and components of the microbial community in soil and bark samples interact with each other. Genera with a relative abundance of less than 1% and a prevalence of less than 50% within each group were excluded from the analysis. Using the “igraph” R package , we thenIn this study, a large-scale characterization of endosphere and rhizosphere microbiomes was performed on peach bark and soil with or without symptoms of gummosis. We found that gummosis was associated with alterations in the bacterial and fungal communities in the bark. Distinctly increased levels of probiotic bacteria and pathogenic fungi are possibly associated with plant health. Despite the absence of a time point for tracing the relationship between gummosis severity and microbial alterations, our pilot study contributed to a better understanding of the microbial ecosystem in peach trees with acute plant disease. In conclusion, our findings propose the academic path of biomarker mining for use in the early diagnosis of gum disease.


Collectively, the replacement of cement with DE could not only improve the concrete strength but also reduce the consumption of cement, thereby lessening the cost of construction as well as indirectly reducing the carbon footprint.Portland cement porous concrete (PCPC) has received immense interest recently due to its environmental aids. Its porous structure helps to reduce the water runoff amount while improving the recharge of groundwater. Earlier studies have concentrated on illustrating and knowing the functional as well as structural properties of PCPC. However, very few studies are available on PCPC in combination with natural silica sources as supplementary cementitious materials (SCMs). Most SCMs are by-products of industrial manufacturing processes and cause some environmental concerns, but with their pozzolanic effect, they could be utilized as partial substitute materials for ordinary Portland cement (OPC) to enhance the strength as well as durability performance. The aim of this study is to evaluate the effects of diatomaceous earth (DE) as a supplementary cementitious material for partial substitution of OPC for Portland cement porous concrete application. Compression strength, split tensile strength, and flexural strength tests were performed to determine the effect of partial replacement. To investigate the impact of test variables, basic tests, including void content and water permeability, were also performed. Compared to the control concrete, the results show that a 15% replacement of cement with DE significantly increased the compressive strength (by 53%) while also providing adequate porosity and better water permeability. Statistical analysis (ANOVA) and regression analysis showed that there is a significant ( Most of the places covered with impervious surfaces, such as concrete, due to urbanization, resulted in groundwater table reduction. Pervious concrete pavements, which are open-graded concrete with interconnecting pores with excellent water permeability and porosity, minimize the issue by allowing the seep-through of rain and stormwater into the ground ,2. The t2 into the atmosphere, which has an adverse effect on the global environment [2  and water to form C-S-H  [Cement, one of the most extensively utilized construction materials, has caused the emission of a considerable amount of COironment ,10. Mostironment ,12,13,14ydrates) , which iydrates) ,17. The ydrates)  and alsoydrates) . Fly ashydrates) . It was ydrates) . The usaydrates) . When ceydrates) . The strydrates) .Diatomaceous earth (DE) is reported to exhibit pozzolanic activity when blended with cement ,26. DE iPrevious studies mainly focused on illustrating and knowing the functional as well as structural properties of PCPC; however, very limited studies evaluated the effects of natural silica sources as supplementary cementitious materials for PCPC application. On the other hand, although the application of DE in concrete and building materials is not a new topic, there are very few studies on the use of DE for Portland cement porous concrete application. The main objective of this study is to improve the understanding of the effect of DE in pervious concrete. In this study, the cement was partially replaced by DE as an SCM. A series of experimental tests, such as consistency and water demand, compressive strength, split tensile strength, flexural strength, void content, and water permeability tests, were conducted on the samples.The production of PCPC includes cement, coarse aggregate, water, and diatomaceous earth as its primary constituents. In this study, the addition of a small fine aggregate (10%) was applied to the mix in terms of improving the strength of the concrete. In addition, the cement was replaced by various amounts of DE.A single source crushed rock having an average particle size of 19 mm was used in this research. The typical grading of coarse aggregate used in PCPC conforms to the specifications of ASTM C33/C33M . Accordi3. Using a JEOL-JSM-IT 200, equipped with EDS, the scanning electron microscopy (SEM) test was carried out. SEM test specimens, in powder forms, were put on platinum-coated carbon adhesive tape and were subjected to characterization examination utilizing a voltage of 15 kV in a high vacuum camper. The phase composition of cement was characterized using the X-ray diffraction (XRD) analysis. The sample was prepared in powder form, put inside a sample holder, and mounted on a triple-axis (Ge-220) monochromatic detector. At room temperature, the sample analysis was carried out utilizing 45 kV, 40 mA, with 2 exploration range. The scanning electron microscope (SEM) and X-ray diffraction (XRD) images of cement are shown in The cement used in this study was commercial ordinary Portland cement (OPC) with the market branded as Coramantal and conformed to ASTM C150/C150M-22, 43 N, , with a Clean tap water was employed for material washing and other experimental purposes. The water quality used in this study was drinkable, as discussed in ACI 301 . The rat3. The specific gravity of DE in standard drying conditions was obtained at 2.06, while it was evaluated at 1.92 after oven drying with absorption of 6.71%. The SEM images of DE at 50 µm and 2 µm are represented in The DE samples were collected from Jaipur, Rajasthan, India. After one day of natural surface drying, the obtained DE sample was crushed and sieved by using a sieve size of 200 μ. Following that, the DE powder was dried in the oven for 24 h at 100 °C. The DE particles showed a bulk density of 775 kg/m3 was used.To find reliable mean values, triplicate specimens were prepared for each test group. The samples were prepared as per ASTM C1688/C1688M . The mix3. Cement was partially replaced by DE by weight at various contents of 5%, 10%, 15%, 20%, and 25%. The different mix ratios are given in The total volume of solids, including the aggregates, water, and cementitious materials, was estimated to be 0.85 mI = ratio of infiltration (mm/h), M = weight of infiltrated water (kg), D = infiltration ring inner diameter (mm), t = time taken for the measured volume of water to infiltrate the concrete (s), and K is 4,583,666,000 ((mm3 s)/(kg·h)).The compressive strength test was performed at the age of 7 days and 28 days by following the ASTM C39  testing 2S, C3S, and C3A 2, while reacting with water, which reacts with alumina and amorphous silica in DE to generate an excessive amount of C-S-H. The excess amount of C-S-H can replace the pore holes in the concrete mix and refine and densify the interfacial transition zone (ITZ) between aggregate particles and cement paste [The average split tensile strength of PCPC specimens on days 7 and 28 is shown in nt paste . The immnt paste . Moreovent paste . Similarnt paste .2 and Ca(OH)2 contributed to the bending resistance, thereby increasing the flexural strength [strength . Saidi estrength . However2 and caused C-S-H gel formation, which helped to fill the pores in the concrete and made the PCPC denser than the control concrete. The addition of DE in cement concrete could reduce the voids. This result is similar to the previous findings of [K = 1,273,240 ((mm3·kg)/(m3·g)), A= weight of dry specimen (g), B = weight of specimen underwater (g), ρw = experimental water density (kg/m3), D = average specimen diameter (mm), and L = average specimen length (mm).As mentioned previously, the volume displacement method was used to determine the amount of void content in each cylindrical sample at the age of 28 days. dings of . HoweverThe values of the permeability coefficient are presented in c = Young’s modulus of concrete, and cF= compressive strength of concrete.p < 0.05) in the physical properties of concrete upon the addition of DE, and the optimum concentration of cement to be replaced by DE was found to be 15% (One-way ANOVA (SPSS 17) was used to find the significance between the control mix and various replacement mixtures. Further, regression analysis was performed to define the relationship between various physical features of PCPC, and the goodness of fit was represented as scatter plots . The reso be 15% . Figure o be 15% B. This mo be 15% C. Finallo be 15% D with thThe diatomaceous earth was observed to be a lightweight SCM having soft minerals with a rich amount of amorphous silica. The SEM image showed that the diatomaceous earth particles have high porosity.It was observed that the water demand gradually increased up to the optimum inclusion of DE, but the higher replacement of cement with DE resulted in a drastic increment in water demand.The strength of pervious concrete containing diatomaceous earth increased in all replacements. The 25% replacement showed only a slightly higher strength than the control mix on the 28th day. However, the 15% replacement, the optimum content, exhibited 54.45%, 36.84%, and 37.84% higher compressive, split tensile, and flexural strengths, respectively, compared to the control mix.The correlation between porosity and permeability showed an inverse relation upon increasing diatomaceous earth concentration. The desired porosity of 15.42% was achieved in 15% replacement. The relationships between Young’s modulus and compressive strength with both experimental and predicted values by partial replacement of cement by diatomaceous earth were almost similar within the range of 10–15%.The addition of diatomaceous earth as an SCM in cement would decrease the amount of cement consumption, contributing to the reduction of carbon footprint in the atmosphere.Although several studies explored the potential use of DE in concrete, there are very limited works on the use of DE for Portland cement porous concrete application. Therefore, the aim of this research was to evaluate the performance of PCPC having cement partially replaced by DE. The study results can be summarized as follows:


Panax notoginseng is an important medicinal plant in China, but there are some limitations in the ecological suitability study, such as incomplete investigation of species distribution, single regionalization modeling, and lack of collaborative evaluation of ecological suitability, and quality suitability. In this study, the maximum entropy model was used to analyze the ecological suitability of P. notoginseng under current and future climates. The multi-source chemical information of samples was collected to evaluate the uniformity between quality and ecology. The results showed that the current suitable habitat was mainly in southwest China. In the future climate scenarios, the high suitable habitat will be severely degraded. Modeling based on different regionalization could predict larger suitable habitat areas. The samples in the high suitable habitat had both quality suitability and ecological suitability, and the accumulation of chemical components had different responses to different environmental factors. Two-dimensional correlation spectroscopy combined with deep learning could achieve rapid identification of samples from different suitable habitats. In conclusion, global warming is not conducive to the distribution and spread of P. notoginseng. The high suitable habitat was conducive to the cultivation of high-quality medicinal materials. Actual regionalization modeling had more guiding significance for the selection of suitable habitats in a small area. The multi-regionalization modeling theory proposed in this study could provide a new perspective for the ecological suitability study of similar medicinal plants. The results provided a reference for the introduction and cultivation, and lay the foundation for the scientific and standardized production of high-quality P. notoginseng. Panax notoginseng (Burk.) F. H. Chen is a perennial herb of Panax genus, whose dry root and rhizome are valuable medicinal herbs, peculiar to China. Modern studies have proved that P. notoginseng mainly contains saponins, flavonoids, volatile oils, sterols, organic acids, polysaccharides, amino acids, and other chemical components  model was used to explore the potentially suitable habitat and simulate the impact of climate change on the distribution of P. notoginseng in the future. At the practical level, modern analytical methods were used to integrate multi-source chemical information of P. notoginseng samples, analyze the ecological suitability regionalization and quality suitability regionalization collaboratively, and establish a rapid identification method of P. notoginseng from high-quality suitable habitat. It is proposed to combine theory and practice on the basis of current species distribution, reveal the geographical distribution pattern of P. notoginseng under current and future climate scenarios, and evaluate the consistency of ecological suitability regionalization and quality suitability regionalization. In this way, the quality of the source is guaranteed, and a scientific basis is provided for the selection of suitable planting regionalization for P. notoginseng under future climate change.The present study conducted a more comprehensive investigation and collection of the actual distribution of P. notoginseng were obtained by three ways: (1) From 2011 to 2018, the Institute of Medicinal Plants, Yunnan Academy of Agricultural Sciences conducted field surveys on the major distribution areas in Yunnan, Guangxi, and surrounding areas, recorded the longitude and latitude information of planting sites in detail, and compiled data sets of species distribution. (2) Species distribution information was obtained through literature review and online sharing platform of specimen digital information. Platforms include the Chinese Virtual Herbarium databases of National Plant Specimen Resource Center , the National Specimen Information Infrastructure , and the Global Biodiversity Information Facility . (3) Government news reports combined with Google satellite images conducted investigation and image analysis of greenhouses for planting P. notoginseng in Yunnan, Guangxi, Guizhou, Sichuan, and in Chongqing from 2020 to 2021, so as to supplement and improve the species distribution data set. Through the integration of the above data sets, 3,475 species distribution data were obtained.The distribution data of 2 grid. Through the above screening, 2,122 species distribution data  were downloaded from the WorldClim (https://www.worldclim.org/). The climate variables required to study current species distributions were obtained from the grid data generated by interpolating observations from global climate stations from 1970 to 2000. Future climate data were predicted by the medium-resolution Climate System Model (BBC-CSM2-MR) developed by the National Climate Center (Beijing). Two greenhouse gas concentration pathways (medium-low concentration scenario: SSP245 and high concentration scenario: SSP585) under two future periods (2050s: 2041–2060 and 2090s: 2081–2100) were selected to study. The precision of future climate data provided by WorldClim is 2.5 min. In order to facilitate modeling, the data of spatial resolution was unified at 30 s by using ArcGIS raster resampling.The environment variables for modeling mainly include elevation data and bioclimatic variables . All theIn order to avoid over-fitting of modeling results due to multicollinearity among environmental variables, elevation and bioclimatic variables corresponding to species distribution data were extracted for Spearman's correlation analysis. Combined with the modeling results, when the variables are significantly correlated and the correlation coefficient is >0.8, the variables with small modeling contributions are eliminated. All environment variables were converted to ASCII format before modeling. The final variables used for modeling are shown in P. notoginseng. Therefore, in addition to modeling for overall regionalization, this study also divided all distribution sites into the following regionalization according to the topographic characteristics of the distribution region, the actual planting situation, the authenticity of planting region, the traditional production area, the new production area, and other restrictions: (1) Hengduan Mountains, (2) Yunnan-Guizhou Plateau, (3) Yunnan + Guangxi, (4) Sichuan Basin, (5) Sichuan + Chongqing + Guizhou, and (6) Sichuan + Chongqing + Guizhou + Guangxi + Wenshan. It is planned to compare and predict the suitable habitat areas of P. notoginseng through different regionalization modeling.Regionalization, which means the division of regions, is a way for people to extract spatial information according to a specific purpose, and to realize the understanding of the natural or the social environment through the region-based classification and merger  value of the area under the receiver operating characteristics (ROC) curve was used to evaluate the accuracy of the model results. The AUC value ranges from 0 to 1, and the closer the value is to 1, the more reliable the prediction results are . The suitability was divided into four categories according to the difference of suitability index: high suitability (HIS ≥ 0.6), moderate suitability (0.4 ≤ HIS < 0.6), low suitability (0.2 ≤ HIS < 0.4), and no suitability (HIS < 0.2)  and two emission scenarios (SSP245 and SSP585) were performed based on the overall regionalization. Then, the HIS for current and future suitable habitats were calculated based on the results of MaxEnt software (probability of species presence). According to the suitability index, the habitat suitability of P. notoginseng was divided into high suitability, moderate suitability, low suitability and no suitability.The spatial extent of suitable habitat of The changes of current and future suitable habitats were evaluated by calculating the suitable habitat change rate (SHCR) of high suitability, moderate suitability, and low suitability.P. notoginseng is mainly concentrated in Yunnan, China. In addition, according to the survey results of actual distribution sites, there are more distribution sites in the five regions of Wenshan, Honghe, Yuxi, Kunming, and Qujing in Yunnan Province. Therefore, considering the actual planting situation, the economic value of production and the prediction results of suitable habitat for the Maxent model, these five regions were selected for sampling, and a total of 455 samples of P. notoginseng were collected. The picture information of some habitats, collection sites, and samples are shown in P. notoginseng (Burk.) F. H. Chen by Professor JinYu Zhang, Institute of Medicinal Plants, Yunnan Academy of Agricultural Sciences. After sampling, the roots of fresh samples were taken, cleaned, and dried in a constant temperature oven at 50°C. Then all samples were crushed with a grinder, screened through 80 mesh, and stored in ziplock bags to avoid light for subsequent analysis. The number of samples used for content analysis and spectral analysis is shown in At present, the cultivation area of 1, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rb1 were determined by high-performance liquid chromatography (HPLC) analysis. The contents of total flavonoids and total polysaccharide were determined by UV-visible spectrophotometry. Chromatographic analyses were performed with LC-10 ATvp liquid chromatograph system with a diode array detector. binary pump, manual sampler, and CLASS-VP workstation. The chromatographic separation was achieved using an Inertsil ODS-3 column . Standards of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, and rutin were purchased from the Chinese National Institute for Food and Drug Control . Anhydrous glucose was purchased from Sigma-Aldrich. HPLC grade acetonitrile and methanol were purchased from Thermo Fisher Scientific . Other chemicals and reagents were of analytical grade.In this study, the contents of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rb1 were precisely measured at 2.00, 3.00, 2.20, and 2.40 mg, respectively, and dissolved in 1 mL of methanol solution. The solution was diluted 7 times according to the gradient of 60% to obtain a total of 8 mass concentration gradients of the reference solution. Sample powder was weighed accurately to 100 ± 0.30 mg and extracted with 2.0 mL of methanol by an ultrasound-assisted method for 60 min at ambient temperature. The methanol loss was supplemented after ultrasound, and the extract solution was filtered using a 0.22 μm membrane filter, which was the solution to be tested. The mobile phase and gradient elution procedures refer to HPLC detection of P. notoginseng in the Chinese Pharmacopeia   to 10 mL. The mother liquor of standard substance was diluted 6 times according to the equal gradient of 60% to obtain the standard substance solution with seven concentration gradients. Sample powder of 50 ± 0.2 mg were accurately weighed, and 8 mL of 70% ethanol solution was added for ultrasonic extraction for 1 h. Then, 6 mL of extracts were absorbed, and color reagent was added to 8 mL. Color developing time was 20 min, and scanning range was 190–600 nm. The background was scanned using a blank solution with chromogenic agent before collecting the sample spectra  of each samples were performed by the Fourier transform infrared spectrometer  equipped with a deuterated triglycine sulfate (DTGS) detector. The sample powder (1.2 ± 0.2 mg) and KBr powder (15.0 ± 1.0 mg) were accurately weighed and mixed in an agate mortar. After grinding, the mixture was poured into a mold and pressed to make uniform flakes. Each sample tablet was tested to obtain FT-MIR spectra that was recorded in the region of 4,000–650 cm−1, and the resolution was set to 8 cm−1. About 64 scans were accumulated for a single sample, and each sample was repeated for three times to obtain the average spectrum. The spectral record after acquisition is the logarithm of reciprocal reflectance.The near infrared spectra (NIR) of sample powder were collected with an Antaris II spectrometer  equipped with an integrating sphere diffused reflection mode and Result 2.1 software. The wavenumber region was set to 10,000–4,000 cmP. notoginseng samples with ecological suitability  has quality suitability, the contents of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, total flavonoids, and total polysaccharides in these samples were compared and analyzed. In addition, we drew the boxplot for intuitively displaying the content data distribution characteristics of P. notoginseng samples from Wenshan, Honghe, Yuxi, Kunming, and Qujing.To investigate whether Principal component analysis (PCA) is an unsupervised pattern recognition method, which transforms multiple related variables into a group of unrelated new variables by dimensionality reduction, while preserving the original sample information as much as possible , where v is the variable and t is the external disturbance , test set (30%), and external validation set (10%) using the Kennard–Stone algorithm.In this study, a 12-layer ResNet was established with a weight attenuation coefficient λ of 0.0001 and a learning rate of 0.01. The AUC value can be used to evaluate the credibility and accuracy of the model results, and its value is between −1 and 1. For the AUC value, 0.60 < AUC ≤ 0.70 indicates that the model effect is fair, the value 0.70 < AUC ≤ 0.80 indicates that the model effect is good, the value 0.80 < AUC ≤ 0.90 indicates that the model effect is very good, and the value 0.90 < AUC ≤ 1.00 indicates that the model effect is excellent Swets, . The AUCP. notoginseng, and the main climate factors affecting the distribution of P. notoginseng were obtained. P. notoginseng with a cumulative contribution rate of 94.08%. The variables of Bio04, Ele, Bio02, Bio17, and Bio14 contributed significantly to the potential distribution of P. notoginseng in Yunnan-Guizhou Plateau regionalization model, with a cumulative contribution rate of 93.40%. For Yunnan + Guangxi regionalization model, Bio04, Bio07, and Bio17 contributed significantly to the potential distribution, with a cumulative contribution rate of 96.48%. The variables of Bio04, Ele, Bio03, and Bio19 had a high contribution rate to the potential distribution in the Sichuan Basin regionalization model, with a cumulative contribution rate of 100.00%. Bio18, Bio14, Bio15, Bio02, Ele, and Bio19 had a high contribution rate to the potential distribution of P. notoginseng in the Sichuan + Guizhou regionalization model, with a cumulative contribution rate of 93.84%. For Sichuan + Guizhou + Guangxi + Wenshan regionalization model, Bio17, Bio04, Ele, Bio13, Bio15, and Bio07 contributed significantly to the potential distribution of P. notoginseng with a cumulative contribution rate of 99.53%.Eleven climate factors were selected as indicators to simulate the suitable habitat of P. notoginseng in different regions. On the whole, the main ecological factors affecting the spatial distribution of P. notoginseng are Bio04, Ele, and Bio17. According to relevant literature reports, the elevation of the most suitable planting area of P. notoginseng is about 1,400–1,800 m, and the incidence of black spot will increase with the higher elevation beyond the suitable range  had significant changes in both area and location. This was caused by the differences of topographic, geomorphic, and climatic factors in different regionalization, which indicated that selecting the suitable habitat in these four divisions should refer to the prediction results of each regionalization rather than the overall regionalization. From models D, E, and F, we can see that Sichuan, Guangxi, and Guizhou should theoretically have larger suitable habitat areas. Since the overall regionalization model is more focused on traditional production areas with more distribution points, it conceals important environmental factors of new production areas, and underestimates the potential and suitability of planting P. notoginseng, and the suitable habitat distribution  were determined , Natural Science Foundation of Yunnan Province of China (Grant Number: 202101AT070260), and Special Program for the Major Science and Technology Projects of Yunnan Province (Grant Number: 202002AA100007).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Descending necrotizing mediastinitis (DNM) is an acute, rare, severe condition with high mortality, but the optimal management protocol is still controversial. We retrospectively analyzed the results of multidisciplinary management in patients treated for DNM at our center over the last twenty years. Fifteen male patients, mean age 49.07 ± 14.92 years, were treated: 9 with cervico-pharyngeal etiopathogenesis, 3 peri-tonsillar/tonsillar, 2 odontogenic, 1 post-surgical; 6 with DNM type I, 6 with type IIA, and 3 with type IIB (Endo’s classification). Mean time between diagnosis and treatment was 2.24 ± 1.61 days. In all cases, mediastinum drainage via thoracotomy was performed after neck drainage via cervicotomy, associated with tooth treatment in two; one required re-operation; tracheostomy was necessary in 9, temporary intensive care unit stay in 4; 6 developed complications, without post-operative mortality. Main isolated germs were Staphylococci and Candida; 7 had polymicrobial infection. The most used antibiotics were meropenem, metronidazole, teicoplanin, third-generation cephalosporins and clyndamicin; anti-fungal drugs were fluconazole, caspofungin and anidulafungin. On multivariate analysis, presence of cardiovascular disease was statistically significantly associated with longer chest tube duration and hospital stay. DNM requires early diagnosis and treatment to reduce mortality and morbidity. The most effective treatment should provide a multidisciplinary approach, combining cervicotomy and thoracotomy to drain all infectious collections with administration and monitoring of the proper antimicrobial therapy. Descending necrotizing mediastinitis (DNM) is an acute and rare serious condition with a high mortality rate (ranging from 11% to 40%) that represents, in most cases, a complication of oro-pharyngeal or cervico-fascial infections, caused by both anaerobic and aerobic bacteria ,3,4,5,6.+ 14.92 years (range 12–70 years). They developed DNM with various etiopathogenesis: 9 had a cervico-pharyngeal origin of infection, 3 peri-tonsillar/tonsillar, 2 odontogenic and one originated as a post-operative complication (hemiglossectomy). Fifteen patients affected by DNM were treated at our center from 1 January 2002 to 31 December 2020; all were males with a mean age of 49.07 The degree of diffusion of DNM according to Endo’s classification, defined using CT scans and intra-operative findings , was disAt the diagnosis of DNM, septic shock was already present in only two patients. In all cases, apart from clinical signs of severe infection, diagnostic certainty was achieved with contrast enhanced neck and chest CT, which allowed precise definition of the location and extent of the infectious process and therefore the best therapeutic strategy in a mean time of 2.24 ± 1.61 days (range 8 h–5 days). In all 15 cases it was necessary to combine a thoracotomy with a cervicotomy to obtain a complete cervical, mediastinal and pleural drainage ; in partOverall, 10 right lateral thoracotomies and 5 left lateral thoracotomies were performed. During thoracotomy, after opening the mediastinal pleura, mediastinal debridement and washing with intra-operative disinfectant solutions was carried out, as well as drainage and toilette of pleural collections and/or aspiration of any collections in other locations; material was collected for microbiological examination and large bore chest tubes were placed. Of the 15 cases analyzed, only one patient required re-operation: in particular, a right lateral re-thoracotomy was performed because of persistent signs of infection and post-operative CT scan evidence of a 7 cm loculated fluid collection with atypical arrangement, at the level of the right apical postero-mediastinal pleura. The mean duration of surgery was 130.63 ± 32.34 min (range 95–195 min). Tracheostomy, performed simultaneously in the thoracotomy/cervicotomy procedure, was necessary in 9/15 (60%) patients and an ICU stay in 4/15 (27%) patients. Post-operative complications occurred in 6/15 (40%) patients: according to the classification of surgical complications by Clavien-Dindo , there wThe mean duration of chest  tubes was 18.91 ± 13.46 days (range 8–52 days).The mean length of hospital stay was 29.33 ± 14.24 days (range 13–61 days) and the outcome of surgical treatment was favorable in all patients, with a post-operative mortality of 0%.Staphylococcus, of which the most frequent were S. epidermidis (4 cases) and S. aureus (2 cases), but there was evidence also of S. hominis (one case) and S. capitis (one case); in 2/15 (13%) cases, bacteria belonging to the genus Streptococcus, in particular S. anginosus and S. cristatus; in 4/15 (27%) cases, bacteria belonging to other genera: K. Pneumoniae, E. coli, S. maltophilia, Serratia marcescens together with P. aeruginosa; in 3/15 (20%) cases, fungi were detected, in particular of the Candida genus: Candida albicans (2 cases) and Candida parapsilosis (one case). In 7/15 (47%) patients, there was a polymicrobial infection and three of them presented with both fungal and bacterial infection. Lastly, 4/15 (27%) patients were negative at microbiological examination. An antibiogram/antimicogram was performed in all positive cases. Regarding the type of germs isolated on the samples taken intra- or post-operatively , we observed, in 8/15 (53%) cases, bacteria members of the genus In all cases, antimicrobial post-operative therapy, on the basis of the antibiogram/antimicogram where available, was established after consultation with the infectious disease specialist, who also monitored the progress of treatment. The most commonly used antibiotics were meropenem, metronidazole, teicoplanin, third-generation cephalosporins and clyndamicin. For the microbiological positivity for fungi on intra-operative samples or blood cultures, anti-fungal drugs were also used, in particular fluconazole, caspofungin and anidulafungin. There were no complications related to the administration of antimicrobial treatments and no therapy had to be discontinued prematurely due to side effects from drugs. The mean body mass index (BMI) was 23.94 ± 2.67 (range 18.66–26.52), which is within the normal weight category. Considering pre-operative risk factors or co-morbidities, we observed smoking and/or COPD in 6/15 (40%) patients, overweight in 4/15 (27%), diabetes mellitus in 3/15 (20%), cardiopathy/vasculopathy in 3/15 (20%), chronic renal failure in 1/15 (7%), chronic liver disease in 1/15 (7%), other diseases in 4/15 (27%). On multivariate analysis, cardiovascular diseases resulted in statistically significant longer chest tube duration and hospital stay.Data relating to the continuous variables considered in our study are summarized in In our study, concerning gender, we found DNM only in males, in agreement with some authors who described a higher incidence in males rather than females ,11, but The mean age was about 50 years and this figure is consistent with the literature ,12, evenReferring to etiopathogenesis, that is the site of origin and source of infection that evolved in DNM, we found an higher frequency for the cervico-pharyngeal (60%) site, followed by the tonsillar/peritonsillar (20%) and odontogenic 13%) sites; an iatrogenic source was found only in one case (7%), due to a hemiglossectomy that became complicated. These findings are only partially in agreement with the literature, because even if they are in line with the most frequent sites of origin  3% sites;,12, mostThe spread of infection was evaluated according to Endo’s classification of DNM ,8: type Septic shock at the time of diagnosis of DNM was present only in 2 13%) patients. This could be explained by the fact that one of them was the oldest of the patients (70-years old), whereas the other one had the most complex clinical history, since he had suffered from ischemic stroke and recurrent syncopal episodes, and was therefore a debilitated patient: therefore, these conditions could have favored the worsening of the infection and the consequent onset of the septic shock. However, this percentage seems to be lower than that reported in other studies, where death has been described after this dangerous complication ,12. Ther% patientThe mean elapsed time between diagnosis of DNM and treatment was about two days, which is very short, similar to what other authors have described ,13. As for the surgical treatment, in all cases we performed thoracotomy after the cervicotomy carried out by otolaryngologysts , with the aim of draining and obtaining a toilette of pleural collections and a mediastinal debridement ; at the same time washing with intra-operative disinfectant solutions was performed. The prevalent thoracic approach was the right one (in 2/3 of cases), through lateral thoracotomy, which we preferred to the postero-lateral one as it is less demanding for the patient in terms of muscular section, ensuring a less painful and faster post-operative recovery for these patients, who were already immunocompromised by DNM. Standard thoracotomy in all its variants has high efficacy to treat DNM, according to many authors and in agreement with our experience ,4,9.In the literature, it is reported that in cases of type I DNM, drainage through cervicotomy is very effective ; howeverA re-operation was necessary in one case (7%) only, namely a right lateral re-thoracotomy, carried out two days after the first operation, due to persistent fever and post-operative CT scan evidence of a 7 cm loculated fluid collection at the level of the apical pleura. This case underlines the importance of CT scan in the follow-up, as the persistence of the infectious process was diagnosed early and treated, thus improving the prognosis of the patient. The mean duration of thoracic surgery was about two hours, and therefore relatively contained, which reduced the risk of intra and post-operative complications; despite this, complications occurred in the post-operative period in 40% of patients, although in all cases they were resolved with conservative treatment.The post-operative complication rate in our study 40%) was similar to that of some authors , but rel0% was siIn our experience, a tracheostomy was necessary in 60% of patients, performed simultaneously in the thoracotomy/cervicotomy procedure, to ensure protection and patency of the airway, which could become obstructed and compressed due to fistulization and edema caused by severe inflammation. To this extent, it is crucial to ensure control of the airway as soon as possible and therefore the patient’s respiratory function, to prevent the onset of serious complications in the context of an already critical situation ,9.Moreover, in 4 patients (27%), a temporary stay in ICU occurred, since two of them were septic already upon arrival at the hospital; one patient, transferred from another hospital, presented with a complicated airway situation with pharyngo-laryngeal swelling and compression of epiglottis, and another patient  developed respiratory failure after cervicotomy and thoracotomy. In fact, this group of patients is more likely to develop life-threatening complications, as reported by Mejzlik et al. . The mean duration of chest  tubes was about 19 days; therefore, in general, drainage remained in place for a long time, as further confirmation of the severity of the disease and of the need for keeping them longer than other types of thoracotomy surgery, as they should be removed after complete resolution of all infectious collections. For this purpose, radiological post-operative monitoring by neck-chest CT scan is essential. Nevertheless, the mean duration of chest tubes in our experience was slightly longer than that reported by other authors like Yanik et al., of about 13 days [The mean time of hospitalization was about 29 days and the outcome of surgery was successful in all patients, with no mortality rate. In this regard, it must be emphasized that hospital stay is quite long due to the severity of the disease, which requires longer recovery times, and that in our experience we had no mortality compared to the literature, reporting mortality rates of about 11–40% , even thStaphylococcus (53%) (most frequently S. epidermidis and S. aureus) and Streptococcus (13%). This is partially in agreement with the literature where in most cases a higher incidence of Streptococci is described [Regarding the type of germs, which were isolated on the samples taken intra-operatively or post-operatively , we predominantly observed positivity for bacteria of the genus ococcus 5%  but this can be explained and justified by the rarity of the disease. Therefore, we recommend multicenter studies in order to have more data available for evaluation and results with statistical significance, especially concerning the identification of prognostic risk factors that may affect the evolution and treatment of this potentially fatal condition.We performed a retrospective study of patients who came at our hospital and were treated for DNM at our Unit of Thoracic Surgery from 2002 to 2020. The DNM diagnosis was made on the basis of the criteria defined by Estrera and colleagues . By retrospective consultation of medical records, we analyzed the following patients’ data: age, gender, etiopathogenesis of DNM (source of infection), degree of diffusion of DNM .Continuous variables were reported as mean with standard deviation. Nominal variables were reported as counts and percentages. Pre-operative risk factors or co-morbidities  were the variables submitted for statistical multivariate analysis, to disclose risk factors for a worse course of DNM . Significance was defined as a DNM is a serious condition that puts the patient’s life at risk and so requires a timely and correct treatment. In most cases, it already presents at an advanced degree of diffusion, thus a rapid diagnosis, with the support of neck and chest CT, plays a fundamental role to reduce the mortality and morbidity of this disease.The standard surgical approach consists in a lateral thoracotomy, which guarantees an optimal exposure and cleaning of the mediastinum and pleural cavity and a lower occurrence of post-operative complications, always preceded by cervicotomy and cervical drainage and eventual treatment of diseased teeth, to treat the source of origin of DNM.In association with surgical treatment, multidisciplinary cooperation with the infectious disease specialist is fundamental to promptly administer the proper antimicrobial therapy, on the basis of the antibiogram/antimicogram where available, and monitor the progress of treatment.Tracheostomy to maintain a patent airway and when required, temporary ICU stay, are essential because delays could have serious consequences. Any recurrence of the disease must be recognized and treated immediately, because delays in re-operation worsen the patient’s prognosis.Ultimately, all factors analyzed in our study should be further evaluated in future multicentric studies, to better understand the expected prognosis of the individual patient and carry out the most appropriate multidisciplinary management and treatment.


Rhododendron species have been paid rare attention in the study of molecular mechanism of tolerance or response to drought. Herein, we investigated the correlation of transcriptomic changes with the physiological and biochemical indicators of Rhododendron rex under drought stress by using the co-expression network approach and regression analysis. Compared with the control treatment, the number of significantly differentially expressed unigenes (DEGs) increased with the degree of drought stress. The DEGs were mainly enriched in the cell wall metabolic process, signaling pathways, sugar metabolism, and nitrogen metabolism. Coupled analysis of the transcriptome, physiological, and biochemical parameters indicated that the metabolic pathways were highly correlated with the physiological and biochemical indicators under drought stress, especially the chlorophyll fluorescence parameters, such as the actual photosynthetic efficiency of photosystem II, electron transport rate, photochemical quenching coefficient, and the maximum quantum efficiency of photosystem II photochemistry. The majority of the response genes related to the metabolic pathways, including photosynthesis, sugar metabolism, and phytohormone signal pathway, were highly expressed under drought stress. In addition, genes associated with cell wall, pectin, and galacturonan metabolism also played crucial roles in the response of R. rex to drought stress. The results provided novel insight into the molecular response of the alpine woody species under drought stress and may improve the understanding of the response of forest ecosystems to the global climate change.Increasing severity of drought stress due to global change and extreme weather has been affecting the biodiversity, function, and stability of forest ecosystems. However, despite being an important component in the alpine and subalpine vegetation in forest ecosystems,  Climate change and extreme weather events have led to rising temperature, and varying precipitation has been widely reported to increase the frequency, magnitude, and spatial extent of drought worldwide . The escPopulus tremula delays photosynthesis and maintains starch reserves under drought stress. Paeonia ostii under drought stress. The stomal conductance and Fv/Fm were also significantly decreased in Brassica rapa under drought stress  in Yulong County, Yunnan Province in June 2020. The collected seedlings were planted in culture pots (19 × 27 × 23 cm3) with ~5.0 kg of soil per pot, cultivated and domesticated with daily normal watering for 2 months in the greenhouse  of Yunnan University, Yunnan Province, China. Then, the drought treatments were conducted according to R. rex was extremely sensitive to water stress, the samples were taken after 3 days of drought treatments (Unpublished data). The leaves were sampled, cleaned, and immediately placed in liquid nitrogen for transcriptomic sequencing.F0), maximum fluorescence from dark-adapted leaf (Fm), Fv/Fm, Y(II), photochemical quenching coefficient (qP), electron transport rate (ETR), and nonphotochemical quenching (NPQ). The photosynthetic characteristics and chlorophyll fluorescence parameters were determined using nine different leaves  for each treatment.Mini-PAM  was used to determine the photosynthetic characteristics. In accordance to 2), and anthocyanin  were measured by the Dualex scientific+. The chlorophyll contents of were assayed according to the method described by a and chlorophyll b were calculated as follows: Chla (mg/L) = 12.72 × A663–2.59 × A645; and Chlb (mg/L) = 22.88 × A645–4.67 × A663. The total soluble sugar was estimated following the anthrone method (w/v) TCA and centrifuged at 3,000 rpm for 10 min. Then, 2 ml of the supernatant was collected from the tube, and 2 ml of 0.67% TBA [prepared in 20% TCA (w/v)] was added. The mixed samples were heated at 95°C for 30 min. After cooling, the absorbance was determined at 450, 532, and 600 nm. The proline content was determined by proline-acid ninhydrin according to The nitrogen balance index (NBI), flavoroids  according to the manufacturer’s instructions. An RNA-seq library was constructed by processing at least 1 μg of RNA per sample into cDNA libraries by using a Truseq RNA sample preparation kit . Then, the libraries were sequenced in an Illumina HiSeq4000 sequencing platform to produce two 150-bp paired-end reads. The quality of the RNA-Seq data was validated by performing RT-qPCR analysis on an ABI7500 fluorescent quantitative PCR machine . The relative expression of each gene was normalized through the 2CYP gene . All priThe raw reads were filtered by discarding the contaminated adaptors by using the default steps in SeqPrep.p-adjust of <0.05 and |log2FC| ≥ 2, and the p values were corrected using the method of Benjamini and Hochberg. To determine the biological functions of the unigene sets, Fisher’s exact test was performed for the GO and KEGG enrichment analysis, and the p value corrected with false discovery rate was <0.05. The data were analyzed on the free online platform of Majorbio Cloud Platform.To identify the differentially expressed unigenes (DEGS) by using DESeq2 , the parR. rex was <0.1. The soft threshold of 6 was selected based on the scale-free topology. Then, the adjacency matrix was converted to the topological overlap matrix. Based on the similarity algorithm, the unigenes were clustered, and the parameter of mergeCutHeight 0.2 was set to define the modules. For each module, the module was evaluated using principal component analysis (PCA), and the first principal component (PC1 is the eigengene for each module) was analyzed by linear regression  to determine the modules associated with the physiological and biochemical indicators. In addition, the correlation of each module and traits were calculated using the Spearman correlation coefficient. The modules were selected (p < 0.05) to perform the GO and KEGG enrichment analysis. The heatmap was visualized by TBtools  was performed. To preprocess and filter the data, the parameter was set to TPM < 2, and the threshold of the variation coefficient of expression in the  TBtools .R. rex showed different changes in the response to drought stress. As expected, the Y(II), ETR, and qP under D2 and D3 treatments were significantly decreased. However, these chlorophyll fluorescence parameters were higher in D1 treatment than in CK , and RT-qPCR data indicated the reliability of the present transcriptomic profiling data , 228 (114 up-regulated and 114 down-regulated), and 1,132 (515 up-regulated and 617 down-regulated) significant DEGs were detected in the D1, D2, and D3 treatments when compared with CK, respectively. The number of DEGs increased with the intensifying degree of drought treatments.R. rex to drought stress. Further analysis showed that the expression levels of genes  involved in the cell wall macromolecular compound metabolic and catabolic process were significantly upregulated in the D3 treatments. Conversely, the genes  involved in pectin and galacturonan metabolic process were significantly downregulated in the D3 treatments  were significantly downregulated in the D3 treatments  involved in the MAPK signaling pathway, plant hormone signal transduction, galactose metabolism, nitrogen metabolic, phenylpropanoid biosynthesis, and amino sugar and nucleotide sugar metabolism were significantly upregulated in the D3 treatments , ETR, and qP, and the black module was significantly correlated with qP . Five trpor, psaD, psb28, petE, and LHCB2, involved in photosynthesis, signaling pathways, and biosynthesis of flavonoids showed a decreasing trend with the intensifying degree of drought treatments , ferredoxin (petF), petJ, and ferredoxin–NADP reductase (petH) were involved in the electron transport process , ETR, and qP. Therefore, the modifications in the expression of the genes involved in photosynthesis would participate in the response of R. rex to drought stress by decreasing the ETR, actual photosynthetic efficiency, qP, and chlorophyll content.Genes involved in photosynthesis, antenna proteins, porphyrin, and chlorophyll were down-regulated under drought stress . The ligeatments , indicat process . The decUSP, INV, and E2.4.1.82) involved in these metabolism processes were upregulated under drought stress in the present study (R. rex subjected to drought stress (USP) and other genes, such as CHIB, gmd, manA, XYL4, RGP, E2.7.1.46, E5.1.3.6, and E3.2.1.14, were also up-regulated in the R. rex under different drought treatments  involved in the biosynthesis of flavonoids were up-regulated under drought stress. The flavonoid contents were also higher in the D2 and D3 treatments than in CK. The results implied that the species regulate the expression of genes related to flavonoid biosynthesis and increase the flavonoid content to response to drought stress. Phyllostachys edulis) under drought stress and characterized its response to drought stress in the field. MDA was generated by the reaction of polyunsaturated fatty acids and oxygen species and could be used to evaluate oxidative damage  were highly expressed under drought stress. Conversely, the genes  involved in pectin and galacturonan metabolism were lowly expressed under drought treatments. The genes of CHIB were not only involved in the cell wall macromolecule catabolic and metabolic processes but also in the MAPK signaling pathway. In addition, CHIB was involved in the amino sugar and nucleotide sugar metabolism. In the MAPK signaling pathway, CHIB was activated and then directed defense responses. Similar to CHIB, E3.2.1.14 was not only involved in the cell wall macromolecule catabolic and metabolic processes but also in the amino sugar and nucleotide sugar metabolism. E3.2.1.14 is a key gene in UDP sugar metabolism. Pectin is mainly composed of the homopolymers of galacturonic acid and maintains proper cell wall structure and function by promoting the binding of Ca2+ with the esterified carboxyl groups of galacturonic acid , ETR, qP, Fv/Fm, chlorophyll content, TSS, proline, and MDA, also participated in the response of R. rex to drought stress. The present study provides new insights into the understanding of the response mechanism of alpine woody plants to drought stress and may enhance the knowledge of the response of forest ecosystems to global climate change.Although numerous studies have been conducted on the response or tolerance to drought stress of crops and model plants, investigations on the mechanism of such response by alpine woody plant are severely deficient. This deficiency will affect the understanding of the response of forest ecosystems to the undergoing global climate change. In the present study, we first identified the transcriptomic, physiological, and biochemical indicators of the response of alpine woody plant The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: National Center for Biotechnology Information (NCBI) BioProject database under accession number PRJNA738298.S-KS, X-LZ, and Y-HW conceived and designed the experiment. X-LZ, J-YM, Z-DL, N-fD, H-QY, and LY performed the experiments. X-LZ, J-YM, Z-DL, N-fD, and LY analyzed the data. S-KS, X-LZ, and Y-HW wrote and approved the manuscript. All authors contributed to the article and approved the submitted version.This study was financially supported by the National Natural Science Foundation of China (31870529 and 31560224), the Major Program for Basic Research Project of Yunnan Province (202101BC070002), the Young Academic and Technical Leader Raising Foundation of Yunnan Province (2018HB035), the Open Fund of Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments (2018DG005), and the Program for Excellent Young Talents.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


This study aimed to explore the clinical application of continuous renal replacement therapy (CRRT) in pediatric patients with acute kidney injury (AKI) after liver transplantation.Pediatric patients who underwent liver transplantation were retrospectively investigated. Those who developed AKI within 1 year after the surgery were included and divided into a CRRT group and a non-CRRT group. The perioperative conditions and postoperative complications of the two groups were compared along with the prognoses of the groups to analyze the high-risk factors of the postoperative CRRT.P < 0.05). Moreover, the proportion of patients in AKI stage 3 is higher in the CRRT group (83.33%) than that in the non-CRRT group (11.11%), P < 0.001. The median time to initiate CRRT was 10 days postoperatively, the median number of CRRT treatments per patient was 2 times, the average duration of each CRRT treatment was 10.1 h, and the average rate of the decrease in blood creatinine per treatment was 25.6%. Results of multivariate logistic regression analysis showed that AKI stage 3 , postoperative unscheduled surgery , and hepatic artery occlusion  were recognized as risk factors for postoperative AKI with CRRT therapy. The one- and two-year survival rates were 72.22% and 72.22% in the CRRT group, respectively; and 97.08% and 96.49% in the non-CRRT group, accordingly. There were statistically significant differences in the one- and two-year survival rates between the two groups (P < 0.001).189 (36.91%) patients developed AKI within 1 year after the liver transplantation surgery. There were 18 patients in the CRRT group and 171 in the non-CRRT group. The differences in the preoperative conditions were not statistically significant between the two groups. Compared with the non-CRRT group, patients in the CRRT group had significantly longer transplantation times, higher volumes of intraoperative hemorrhage, and increased incidence of postoperative unscheduled surgery, postoperative primary nonfunction of the transplanted liver, secondary liver transplantation, hepatic artery occlusion, and intestinal fistula (The incidence of AKI after liver transplantation in pediatric patients was high. Patients with AKI stage 3, hepatic artery occlusion, and underwent unscheduled surgery postoperatively were with a high likelihood of receiving CRRT, which was related to a lower one- and two-year survival rates. CRRT effectively improved the one- and two-year survival rates. Liver transplantation is the largest abdominal surgery. The vast intraoperative hemodynamic fluctuations and sizeable intraoperative blood loss/transfusions involved have a significant impact on systemic circulation and renal perfusion, resulting in a high incidence of acute kidney injury (AKI) after liver transplantation. Most pediatric liver transplant recipients are infants and young children under the age of 3 years; their organ functions are not well developed, and they have poor compensatory capacity. Therefore, the incidence of AKI after liver transplantation in children is high, with a reported incidence of 40% to 70% . AccordiContinuous renal replacement therapy (CRRT) is one of the most commonly used forms of blood purification, which can remove toxins and inflammatory medium and small molecule mediators, making it useful for treating renal failure, heart failure, volume overload, sepsis, multiple organ dysfunction syndromes (MODS), adult respiratory distress syndrome (ARDS), severe pancreatitis, and some immune diseases. However, there are few studies on the risk factors and prognosis of CRRT in pediatric patients with postoperative AKI after the liver transplantation.This study retrospectively investigated the pediatric patients with postoperative AKI and CRRT after liver transplantation, analyzed their risk factors and the characteristics of the CRRT, and discussed the prognosis of the involved pediatric patients.A retrospective analysis was conducted on pediatric patients who underwent liver transplantation at Tianjin First Central Hospital from January 1, 2019, to June 1, 2021, and developed AKI within 1 year after the surgery. According to the criteria for AKI established by the organization Kidney Disease: Improving Global Outcomes, those with an absolute increase in blood creatinine (Cr) of ≥26.5 mmol/L within 48 h, or an increase of ≥50% from the baseline, or a urine volume of <0.5 ml/(kg/h) , were considered to be AKI stage 1. Those with an elevation in blood Cr of ≥2-fold from the baseline or a urine volume of <0.5 ml/(kg/h) (≥12 h) were considered to be AKI stage 2. Those with a blood Cr level of ≥353.6 mmol/L, or ≥ an elevation of 3-fold from the baseline, or a urine volume of <0.3 ml/(kg/h) (≥24 h) were considered to be AKI stage 3, in which case, renal replacement therapy could be initiated.The exclusion criteria were pediatric patients who had preoperative renal insufficiency or had undergone renal replacement therapy preoperatively.The present study conformed to the standards of the Ethics Committee of Tianjin First Central Hospital . The guardians of the pediatric patients were informed, and written consents were obtained.Pediatric patients with postoperative AKI after the liver transplantation were divided into CRRT and non-CRRT groups according to whether the CRRT was conducted or not. Before the initiation of the CRRT treatment, no pediatric patients had undergone the liver bridging therapy such as molecular absorbent recirculating system (MARS), single-pass albumin dialysis (SPAD), and plasma separation.The details of the CRRT are as follows:Regardless of whether the patient was diagnosed with AKI stage 3 or not, The CRRT therapy was initiated when patients were with persistent deterioration of renal function after the liver transplantation, with the failure of active treatment of the potential causes, such as reversing systemic hypovolemia and renal hypoperfusion and replacing the use of nephrotoxic drugs, and with symptoms such as anuria, severe hyperkalemia, severe metabolic acidosis, acute pulmonary edema, lethargy, and disturbance of consciousness occurred for a consecutive 48-h.The indwelling of double-lumen catheters was performed under ultrasound guidance. Low-dose sedatives and analgesics were administered before the catheter indwelling procedure to reduce agitation, pain, and stress during puncture. The left internal jugular vein was selected for the indwelling double-lumen catheter in most of the pediatric patients. A Campbell 6F or 8F double-lumen catheter was selected for puncture according to the height and weight of the pediatric patient.−1/min−1, the total flow rate of the dialysis solution or replacement solution was 50–80 ml/kg−1/h−1, and the treatment time was 6–24 h.The mode of CRRT was decided based on the hepatic and renal function as well as the inflammatory response of the pediatric patients. If a patient was predominantly in renal failure, hemodialysis was the preferred method of dialysis to remove toxins with a small molecular weight. If the patient had hepatic dysfunction, a significant systemic inflammatory response, and was less hemodynamically stable, hemofiltration was considered to remove toxins and inflammatory mediators with small or medium molecular weights. Hemodiafiltration was an option if the pediatric patient had renal failure combined with sepsis and severe inflammatory response. A model IQ-21  dialysis machine, a CHDF-21P blood pipeline , and an AEF-03  hemofilter were used. During treatment, the blood flow rate was 3–5 ml/kg*10∧9/L, 3–5 mg/h of heparin sodium was administered for anticoagulation. For those with no bleeding tendency, good coagulation recovery, and a platelet count of <10*10∧9/L, 1–2 mg/h of argatroban was administered for anticoagulation. If the patient had a bleeding tendency, a sodium citrate anticoagulant was administered in a local manner at a flow rate of 1.2–1.5 times the blood flow rate/h. During treatment, the international normalized ratio, kaolin partial thromboplastin time, activated clotting time, and calcium concentration in the arterial and post-filtered blood were monitored depending on the anticoagulant used.If the child had no bleeding tendency with well-recovered coagulation function and the platelet count was >10If the child weighed <10 kg or had hypotension or insufficient effective circulating blood volume, 10 g of albumin was administered to prefill the pipeline. The blood was drained slowly at a low rate, especially during the first 30 min of treatment, and the changes in vital signs were closely monitored. For those with underlying heart disease or heart failure symptoms, slow, little, or even no blood return was required at the end of treatment to prevent the induction of heart failure.The patients' blood urea nitrogen, Cr, cystatin, B-type natriuretic peptide precursor, and blood gas analysis were monitored before and after each treatment. In cases of improved renal function, gradually increased urine volume, reduced inflammatory reaction, no obvious acidosis or hyperkalemia with stable circulation, and no obvious fluid overload performance together with gradually improved functions of the heart, liver, and other organs, the duration of each blood purification treatment was reduced and the interval of blood purification extended until the treatment was terminated.There was no consensus or other common indications for determining when the CRRT should be discontinued temporarily or permanently, so the duration of each CRRT treatment and the total number of CRRT treatments were determined by the responsible pediatrician according to the clinical situation of the patient. The duration of each CRRT treatment plan was arranged for 6 to 12 h, and the final duration of each treatment was determined according to the tolerance of the child, the maintenance of the dialyzer, and the achievement of the primary treatment goals .s), while those that failed to satisfy the normal distribution were expressed as Median . The countable data were expressed as percentages (%). For comparison between groups, a t-test or rank-sum test for independent samples was used for the measurement data, and Fisher's exact probability test or Fisher's exact probability method was used for the categorical variables. The significant levels in all the statistical tests were set as P < 0.05. CRRT-related variables with statically significant differences were included in a multivariate logistic regression analysis to identify the risk factors by enter method, the results were expressed as , and variables with P < 0.05 were set as a statistically significant difference.The study used SPSS 19.0 software for data processing. For the measurement data, a normal distribution test was performed first. The normally distributed measurement data were expressed as mean ± standard deviation  was calculated.P < 0.001), higher volumes of intraoperative hemorrhage , and a higher proportion of patients in AKI stage 3 .Of the 512 pediatric cases with liver transplantation conducted at Tianjin First Central Hospital from January 1, 2019, to June 1, 2021, 189 cases (36.91%) developed AKI within 1 year, mostly within 2 weeks postoperatively , and gradually returned to preoperative levels within 1 month postoperatively . These pediatric patients with postoperative AKI were divided into a CRRT and a non-CRRT group according to whether they were treated with CRRT or not. The general preoperative and intraoperative conditions between the two groups of pediatric patients are shown in P < 0.001), postoperative primary nonfunction of the transplanted liver , secondary liver transplantation , hepatic artery occlusion , and intestinal fistula .Comparisons of the postoperative complications are presented in Of the 189 pediatric liver transplant recipients with AKI occurring between January 1, 2019, and June 1, 2021, 18 were treated with CRRT. These pediatric patients were all indwelled with a double-lumen catheter under ultrasound guidance. The main indwelling site was the left internal jugular vein (66.66%), followed by the right internal jugular vein (33.33%). Continuous renal replacement therapy was conducted a total of 59 times in the 18 patients. Among them, 25 (46.30%) involved continuous venovenous hemodiafiltration, 18 (33.33%) involved continuous venovenous hemodialysis, and 13 (24.07%) involved continuous venovenous hemofiltration. Among these 18 patients received CRRT, seven patients with AKI directly (0–7 days) after primary liver transplantation (38.89%), eight patients developed AKI later but <30 days after primary transplantation (44.44%), and three developed AKI later than 30 days (16.67%). The anticoagulation of the pipeline and filter was dominated by sodium citrate (59.26%), argatroban (25.93%), and sodium heparin (14.81%). The completion rate of CRRT was 96.61%, with only two treatments being terminated due to septic shock with circulatory instability. The median time to initiate CRRT was 10 days postoperatively. Pediatric patients who underwent unscheduled surgery started CRRT an average of 1.5 (0–15) days after their unscheduled surgery; the median number of CRRT treatments per patient was 2 (1–14), the median duration of each treatment was 10.1 (6–19.3) h, and the average rate of decrease in blood Cr per treatment was 25.6% (13.5%-45%). The therapeutic conditions in pediatric patients with AKI are illustrated in P = 0.016], postoperative unscheduled surgery , and hepatic artery occlusion  were recognized as risk factors for postoperative AKI with CRRT therapy (Variables with significant differences in  therapy .P < 0.001). The survival curves of the two groups are plotted in Until October 1, 2021, 189 pediatric patients with AKI were followed up for 4 to 33 months (average follow-up duration: 9.5 months). Thirteen of the children with AKI in the CRRT group improved, and five died. The causes of death were antibody-mediated rejection, sepsis, acute left heart failure, septic shock, and massive gastrointestinal hemorrhage. The time of death ranged from 13 to 58 days postoperatively. In the non-CRRT group, six pediatric patients with AKI died. The causes of death were abdominal infection 2 weeks after liver transplantation in two cases, septic shock 1 month after surgery in two cases, severe pneumonia 6 months after surgery in one case, and lymphoma 24 months after surgery in one case. The one- and two-year survival rates were 72.22% and 72.22% in the CRRT group, respectively, and 97.08% and 96.49% in the non-CRRT group, accordingly. There were statistically significant differences in the one- and two-year survival rates between the two groups . Postoperative unscheduled surgery was caused mainly by serious complications, such as intestinal perforation, abdominal hemorrhage, portal thrombosis, nonfunction of the transplanted liver, etc. These resulted in abdominal infection, hypotension, anemia, acidosis, disturbance of the internal environment, sepsis, and even shock, which further aggravated AKI. It was noted that 8 of the 18 patients in the CRRT group had a second surgery after liver transplantation, and the median time to initiate CRRT for these patients was 1.5 (0 to 15) days after the second surgery. These results suggested that severe postoperative complications and the blow of a secondary operation would accelerate impaired renal function and drive CRRT.Current studies on postsurgical AKI have focused on the influence of preoperative and intraoperative factors on the development of AKI, while less research has been conducted on the association of postoperative factors with the prognosis of AKI. Based on a previous study by the anesthesiologists at our hospital, a retrospective analysis of 512 infants and younger children with liver transplantation was conducted, focusing on the development of AKI after liver transplantation, the postoperative complications, and the prognosis of AKI in pediatric patients. The statistical results revealed that a total of 189 pediatric patients developed AKI within 1 year after surgery with an incidence of 36.91%, which was close to that reported in the literature. These patients were divided into CRRT and non-CRRT groups according to whether or not CRRT was conducted. There was no significant difference between the two groups in terms of preoperative PELD score, antimicrobial exposure, and the indwelling of a central venous catheter. In terms of intraoperative factors, the CRRT group had a longer operation time and a larger volume of intraoperative hemorrhage. In terms of postoperative factors, the incidence of postoperative unscheduled surgeries, primary nonfunctions of the transplanted liver, secondary liver transplantations, hepatic artery occlusions, and intestinal fistulas were higher in the CRRT group than the non-CRRT group  and State Natural Science Fund Project (82170672).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The accuracy and precision of analyte concentrations measured in whole blood by chemiluminescence immunoassay (CLIA) have been significantly affected by erythrocytes, which leads to poor application of whole blood CLIA in clinical practice. In this work, a chemiluminescence biosensing optical platform for blood hematocrit (HCT) analysis using MAGICL 6000  was designed, implemented, and fully characterized. The developed method was successfully applied to determine various HCT levels of human blood from 0% to 65%, with a correlation coefficient of 0.9885 compared with the conventional method . A mathematical model was developed to quantitatively evaluate the impact of HCT on the results of two sample types (whole blood vs. plasma). Combining the established HCT method and mathematical model with CLIA on MAGICL 6000, the precision was significantly improved by almost 20%. Comparison studies using whole blood samples and corresponding plasma samples showed that the square of the correlation coefficients of troponin I (cTnI), myoglobin (MYO), creatine kinase MB (CK-MB), and N-terminal pro-hormone brain natriuretic peptide (NT-proBNP) were increased to 0.9992, 0.9997, 0.9996, and 0.9994, respectively, showing a great potential for clinical application. The ratio of the packed red blood cell volume to the total volume of blood, known as hematocrit , provides critical information about a patient’s health status that helps physicians to diagnose and treat various medical conditions, ailments, and diseases ,2,3. TheChemiluminescence immunoassay (CLIA) combined a chemiluminescent system with immunoreaction is one of the most widely used biomedical diagnostic methods due to its sensitivity and specificity ,11,12,13Troponin I (cTnI), myoglobin (MYO), and creatine kinase MB (CK-MB) assays provide enhanced detection of myocardial injury and potentially improve the efficiency of the diagnosis and management of acute myocardial infarction (AMI) patients . Studies® MAGICL 6000 was investigated.POCT techniques are currently investigated and applied in many fields ,30,31,32® MAGICL 6000 is a fully automated piece of diagnostic equipment that uses CLIA technology to determine the presence of antigens, antibodies, and analytes in samples. The reactants necessary to perform CLIA assays include paramagnetic microparticles coated with a capture molecule specific for the analyte being measured and acridinium-labeled conjugate used to form an immunocomplex sandwich. The trigger  and pre-trigger (peroxide) solution create an oxidative reaction when combined with acridinium conjugate. The specimens, including human whole blood, plasma, and serum, were tested and found acceptable for a variety of myocardial marker detections such as cTnI, MYO, CK-MB, and NT-proBNP on the analytical instrument. With whole blood samples, the analyzer’s reader subsystem measured HCT at each sampling. HCT correction was calculated by a software algorithm, and finally, the results were reported accurately in whole blood. The biological principles of the procedure are as follows: a certain volume of whole blood samples was lysed with the denaturing agents containing 20 mmol/L CTAB and 1 mol/L ammonium chloride (no pretreatment was required for serum or plasma), and then incubated at 37 °C with microparticles coated capture antibodies and acridinium-labeled detection antibodies. After 7 min incubation, the resulting immunocomplexes were separated, washed to remove unbound substances, and then transferred to an optical detection vessel for reading with the MAGICL 6000 counting detection system. The results are determined via a calibration curve, which is instrument-specifically generated by two-point calibration and a master curve provided via the reagent barcode. The commercial cTnI, MYO, CK-MB, NT-proBNP CLIA kit was provided by Getein Biotechnology. The chemicals were purchased from Sigma-Aldrich  at analytical reagent grade, and deionized, doubly distilled water was used throughout.GeteinThe blood samples of 58 healthy people used for HCT analysis by a chemiluminescence biosensor and 86 AMI patients used for myocardial markers immunoassay were obtained from Xuzhou Central Hospital, and approved by Xuzhou Central Hospital Research Ethics Committee. To evaluate the potential impact of the red blood cells on CLIA, plasma with EDTA-K2 as an anti-coagulant and the matched whole blood were used. The EDTA-K2 plasma samples were used after centrifugation and whole blood samples were used within 2 h.Multiple methods exist for the determination of HCT in the clinical laboratory. However, the chemiluminescence biosensor method has not been reported. A total of 100 mL collected whole blood was transferred to eight 10 mL polypropylene transparent tubes and manually reconstituted to eight different HCT levels of 0, 25, 30, 35, 40, 45, 50, 55, and 60% by either adding plasma to the centrifuged whole blood samples or removing it. A desired range of HCTs was prepared to generate the standard calibration curve. The chemiluminescence method on the MAGICL 6000 analyzer was as follows, after adding 2 μL of the prepared whole blood sample into the reaction cup, the pre-trigger solution containing 0.5% W/V hydrogen peroxide and 0.2 mol/L nitric acid, and the trigger solution containing 0.35 mol/L sodium hydroxide and 0.2% W/V CTAC were added to excite heme in blood cells as the substrate; the resulting chemiluminescent reaction was measured as relative light units (RLUs). The excitation conditions were consistent with the process of acridinium conjugate excitation in immunoassay. A direct relationship exists between the amount of red blood cells in the sample and the RLUs detected by the photomultiplier tube optical system.To assess the effect of potentially interfering substances on hematocrit measurement, a phosphate buffer solution as a negative sample and four potentially interfering substances at high concentrations  were added in fixed ratios  to generate the samples containing gradient concentrations of interfering substances. The whole testing was performed in duplicate, and the interference was evaluated by calculating the deviation of the RLUs between potential interferent-spiked samples and unspoked  samples. No interference was defined as ≤10% difference for the average of all RLUs tested.In order to evaluate the proposed method as a platform for the rapid determination of HCT in patient samples, 58 EDTA-treated whole blood samples were collected by convenience sampling depending on research staff and sample availability. Following sample mixing, HCT was measured both on the MAGICL 6000 analyzer by a chemiluminescence biosensor, as mentioned above, and on the Sysmex XE 5000 by impedance according to the manufacturer’s instructions, respectively, within a maximum of 2 h.The relationship between HCT and analyte concentration in the whole blood and plasma was described in a previous literature report and is shown in Equation (1) ,13,19. TTo investigate the influence of the HCT, reconstituted blood samples at different HCT levels of 0, 25, 30, 35, 40, 45, 50, 55, and 60% consistent with the above preparation methods were gently mixed with cTnI standard using a tube shaker to disperse the analyte evenly within the blood samples. cTnI was added in five concentration gradients ranging from 0.1 to 50 ng/mL. The 45 samples prepared were tested on a MAGICL 6000 analyzer for deviation analysis.® CLIA cTnI, MYO, CK-MB, NT-proBNP reagents. The mean, SD, and CV% were calculated.The repeatability study was assessed with the patient specimens following CLSI guideline EP5-A3 on the MAGICL 6000 chemiluminescence analyzer. Levels 1–3 of three whole blood samples with plasma equivalent levels were assayed 10 times to assess the repeatability by the GeteinPlasma and the whole blood sample types were collected from each patient to evaluate the potential impact of the blood cells and HCT correction on the measured concentration of CLIA. Eighty-six paired samples with detectable myocardial marker concentrations obtained from AMI patients were analyzed using the MAGICL 6000 assays. cTnI, MYO, CK-MB, and NT-proBNP were measured in two sample types within 2 h following collection. The results in the whole blood samples were obtained after software correction for individual HCT values according to the correction formula. The regression equation and correlation coefficient were performed to evaluate the correlation between the two sample types, and the mean relative error was estimated by the Bland–Altman method to determine the accuracy of the whole blood samples.p < 0.05.With Microsoft Excel 2007, the results were tabulated. Method comparisons were performed using MedCalc software (v.19.5.6) via Passing Bablok regression and the Bland–Altman plot. Statistical analysis on a completely randomized design was conducted using the one-way analysis of variance (ANOVA) procedure with SPSS software (v.23.0). Numerical data were expressed as mean ± standard deviation (SD). Measured values from plasma samples were compared with those obtained from the whole blood samples using a paired-sample t-test. Statistical significance was considered at the level of To characterize the correlation between the blood HCT and chemiluminescent intensity triggered by hydrogen peroxide and sodium hydroxide solutions, the reconstituted blood samples at different HCT levels of 0, 25, 30, 35, 40, 45, 50, 55, and 60% A were prTo determine the specificity of the HCT assay, endogenous interfering substances were investigated. No significant interferences were observed with human serum albumin concentrations at 500.0 mg/dL; triglycerides at 1000.0 mg/dL; bilirubin at 50.0 mg/dL; and total cholesterol at 2000.0 mg/dL. To evaluate the accuracy of the chemiluminescence biosensor method, the HCT analysis of EDTA-whole blood samples was compared with HCT values determined on a Sysmex XE 5000 analyzer with electronic resistance detection in the clinical lab. A method comparison for HCT testing was performed by Passing Bablok regression and the Bland–Altman analysis method, and the results are shown in p > 0.05) compared with the plasma sample (HCT = 0%), indicating that the HCT value had a strong influence on cTnI analysis. After correction by Equation (1), the accuracy was effectively improved for HCT between 25% and 40%. However, the correction formula was not applicable for higher HCT values, indicating that the difference in cTnI concentration in high HCT samples was not only due to the reduction of available plasma volume, but also due to the reduction of excessive cell debris in the reaction system, which slowed the rate of immune binding. The HCT value is discussed as a factor influencing the analyte concentrations of the immunoassay. Therefore, blood samples with HCT values ranging from 0 to 60% were evaluated for their effect on cTnI quantification. In detail, one blood sample at the HCT of 43% containing an average cTnI concentration of 0.125 ng/mL was adjusted to eight different HCT values  and analyzed. The results of the HCT effect for cTnI CLIA detection are shown in As shown in The analytical performances of cardiac markers including cTnI, MYO, CK-MB, and NT-proBNP were evaluated on a MAGICL 6000 chemiluminescence analyzer. For precision analysis, three whole blood samples were tested 10 times with or without HCT correction on the days of sample collection. Their average concentrations were 0.114, 0.522, and 5.471 ng/mL for cTnI, 120.2, 588.1, and 2023.1 ng/mL for MYO, 10.1, 52.8, and 155.0 ng/mL for CK-MB, and 128.1, 5152.6, and 21006.8 pg/mL for NT-proBNP, respectively. As shown in The concentrations of myocardial markers in the whole blood samples and corresponding plasma samples were compared to assess the potential effect of red blood cells and HCT correction on accuracy. As shown in In this work, the rapid quantification of blood HCT was successfully completed by a chemiluminescence biosensor using a CLIA-based platform. To demonstrate the accuracy of this technique, 58 clinical samples were analyzed in our laboratory with the results correlating very well with the clinical analyses (Sysmex XE 5000). The study of HCT influence on cTnI immunoassay indicated that increased HCT impaired immune binding when analyzed in whole blood. With Equation (2) established by a four-parameter logistic curve fit method, the recovery in the whole blood sample was significantly improved. Combining this HCT measurement and correction with CLIA, which shares the same excitation conditions and photon acquisition device on the MAGICL 6000, we can directly detect the myocardial marker concentrations in the whole blood samples without the need for other HCT measurement instruments. According to the performance analysis, the established method has excellent reproducibility and accuracy due to the advantages of HCT correction, which avoids the influence of erythrocyte inhomogeneity on repeated sampling and excessive cell debris on immune binding.