Prescription optimization is a crucial aspect in the study of traditional Chinese medicine （TCM） compounds. In recent years， the introduction of mathematical methods， data mining techniques， and artificial neural networks has provided new tools for elucidating the compatibility rules of TCM compounds. The study of TCM compounds involves numerous variables， including the proportions of different herbs， the specific extraction parts of each ingredient， and the interactions among multiple components. These factors together create a complex nonlinear dose-effect relationship. In this context， it is essential to identify methods that suit the characteristics of TCM compounds and can leverage their advantages for effective application in new drug development. This paper provided a comprehensive review of the cutting-edge optimization experimental design methods applied in recent studies of TCM compound compatibilities. The key technical issues， such as the optimization of source material selection， dosage optimization of compatible herbs， and multi-objective optimization indicators， were discussed. Furthermore， the evaluation methods for component effects were summarized during the optimization process， so as to provide scientific and practical foundations for innovative research in TCM and the development of new drugs based on TCM compounds.

ObjectiveTo investigate the effects of Dihuang Yinzi on the cognitive function in the mouse model of learning and memory impairments induced by scopolamine （SCOP） and explore the treatment mechanism. MethodsA mouse model of learning and memory impairment was induced by intraperitoneal injection of SCOP. Sixty male C57BL/6J mice were randomized into six groups： control （0.9% NaCl， n=10）， model （SCOP 1 mg·kg^-1·d^-1， n=10）， low-， medium-， and high-dose Dihuang Yinzi （SCOP 1 mg·kg^-1·d^-1 + Dihuang Yinzi 5.5， 11.0， and 22.0 g·kg^-1·d^-1， n=10）， and donepezil （SCOP 1 mg·kg^-1·d^-1 + donepezil 0.84 mg·kg^-1·d^-1， n=10）. Mice were administrated with corresponding drugs for 6 weeks. Modeling started in the 4th week， and mice in other groups except the control group were injected with SCOP intraperitoneally 40 min after daily gavage. Behavioral testing began in the 5th week， 30 min after modeling each day. The Morris water maze and novel object recognition tests were carried out to evaluate the spatial learning and memory function of mice. Nissl staining was employed to observe the survival of neurons and Nissl bodies in the hippocampal CA1 region. Western blot was employed to determine the protein levels of silent information regulator 2 （SIRT2）， α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid （AMPA） receptor 1 （GluA1）， protein kinase A （PKA）， cAMP response element-binding protein （CREB）， phosphorylated-CREB （p-CREB）， postsynaptic density protein 95 （PSD95）， growth-associated protein-43 （GAP-43）， and synaptophysin （SYN） in the hippocampus. Immunofluorescence was used to detect the expression of doublecortin （DCX） in the hippocampal dentate gyrus （DG） region. ResultsCompared with the control group， the model group showed impaired learning and memory （P<0.01）， obvious neuronal damage in the hippocampal CA1 region， a reduction in neuron survival （P<0.01）， a decrease in DCX expression in the hippocampal DG region （P<0.01）， down-regulated proteins levels of GluA1， PKA， p-CREB/CREB， PSD95， SYN， and GAP-43 in the hippocampal tissue （P<0.05， P<0.01）， and an up-regulated protein level of SIRT2 （P<0.01）. Compared with the model group， the medium- and high-dose Dihuang Yinzi groups and the donepezil group showed improvements in learning and memory （P<0.05， P<0.01）， while the low-， medium-， and high-dose Dihuang Yinzi groups and the donepezil group had increased neuron survival （P<0.05， P<0.01）. The medium-dose Dihuang Yinzi group and the donepezil group showed increased DCX expression （P<0.05， P<0.01）. The medium- and high-dose Dihuang Yinzi groups and the donepezil group showed up-regulation in the protein levels of GluA1， PKA， p-CREB/CREB， PSD95， SYN， and GAP-43 （P<0.05， P<0.01） and down-regulation in the protein level of SIRT2 （P<0.01）. ConclusionDihuang Yinzi can improve the cognitive function in the mouse model of learning and memory impairments induced by SCOP by inhibiting the upregulation of SIRT2， activating the PKA/CREB signaling pathway， improving synaptic plasticity， and reducing hippocampal neuronal damage.

Collagen is a major structural protein in the matrix of animal cells and the most widely distributed and abundant functional protein in mammals. Collagen’s good biocompatibility, biodegradability and biological activity make it a very valuable biomaterial. According to the source of collagen, it can be broadly categorized into two types: one is animal collagen; the other is recombinant collagen. Animal collagen is mainly extracted and purified from animal connective tissues by chemical methods, such as acid, alkali and enzyme methods, etc. Recombinant collagen refers to collagen produced by gene splicing technology, where the amino acid sequence is first designed and improved according to one’s own needs, and the gene sequence of improved recombinant collagen is highly consistent with that of human beings, and then the designed gene sequence is cloned into the appropriate vector, and then transferred to the appropriate expression vector. The designed gene sequence is cloned into a suitable vector, and then transferred to a suitable expression system for full expression, and finally the target protein is obtained by extraction and purification technology. Recombinant collagen has excellent histocompatibility and water solubility, can be directly absorbed by the human body and participate in the construction of collagen, remodeling of the extracellular matrix, cell growth, wound healing and site filling, etc., which has demonstrated significant effects, and has become the focus of the development of modern biomedical materials. This paper firstly elaborates the structure, type, and tissue distribution of human collagen, as well as the associated genetic diseases of different types of collagen, then introduces the specific process of producing animal source collagen and recombinant collagen, explains the advantages of recombinant collagen production method, and then introduces the various systems of expressing recombinant collagen, as well as their advantages and disadvantages, and finally briefly introduces the application of animal collagen, focusing on the use of animal collagen in the development of biopharmaceutical materials. In terms of application, it focuses on the use of animal disease models exploring the application effects of recombinant collagen in wound hemostasis, wound repair, corneal therapy, female pelvic floor dysfunction (FPFD), vaginal atrophy (VA) and vaginal dryness, thin endometritis (TE), chronic endometritis (CE), bone tissue regeneration in vivo, cardiovascular diseases, breast cancer (BC) and anti-aging. The mechanism of action of recombinant collagen in the treatment of FPFD and CE was introduced, and the clinical application and curative effect of recombinant collagen in skin burn, skin wound, dermatitis, acne and menopausal urogenital syndrome (GSM) were summarized. From the exploratory studies and clinical applications, it is evident that recombinant collagen has demonstrated surprising effects in the treatment of all types of diseases, such as reducing inflammation, promoting cell proliferation, migration and adhesion, increasing collagen deposition, and remodeling the extracellular matrix. At the end of the review, the challenges faced by recombinant collagen are summarized: to develop new recombinant collagen types and dosage forms, to explore the mechanism of action of recombinant collagen, and to provide an outlook for the future development and application of recombinant collagen.

ObjectiveIn the realm of forensic science, dust is a valuable type of trace evidence with immense potential for intricate investigations. With the development of DNA sequencing technologies, there is a heightened interest among researchers in unraveling the complex tapestry of microbial communities found within dust samples. Furthermore, striking disparities in the microbial community composition have been noted among dust samples from diverse geographical regions, heralding new possibilities for geographical inference based on microbial DNA analysis. The pivotal role of microbial community data from dust in geographical inference is significant, underscoring its critical importance within the field of forensic science. This study aims to delve deeply into the nuances of fungal community composition across the urban landscapes of Beijing, Fuzhou, Kunming, and Urumqi in China. It evaluates the accuracy of biogeographic inference facilitated by the internal transcribed spacer 2 (ITS2) fungal sequencing while concurrently laying a robust foundation for the operational integration of environmental DNA into geographical inference mechanisms. MethodsITS2 region of the fungal genomes was amplified using universal primers known as 5.8S-Fun/ITS4-Fun, and the resulting DNA fragments were sequenced on the Illumina MiSeq FGx platform. Non-metric multidimensional scaling analysis (NMDS) was employed to visually represent the differences between samples, while analysis of similarities (ANOSIM) and permutational multivariate analysis of variance (PERMANOVA) were utilized to statistically evaluate the dissimilarities in community composition across samples. Furthermore, using Linear Discriminant Analysis Effect Size (LEfSe) analysis to identify and filter out species that exhibit significant differences between various cities. In addition, we leveraged SourceTracker to predict the geographic origins of the dust samples. ResultsAmong the four cities of Beijing, Fuzhou, Kunming and Urumqi, Beijing has the highest species richness. The results of species annotation showed that there were significant differences in the species composition and relative abundance of fungal communities in the four cities. NMDS analysis revealed distinct clustering patterns of samples based on their biogeographic origins in multidimensional space. Samples from the same city exhibited clear clustering, while samples from different cities showed separation along the first axis. The results from ANOSIM and PERMANOVA confirmed the significant differences in fungal community composition between the four cities, with the most pronounced distinctions observed between Fuzhou and Urumqi. Notably, the biogeographic origins of all known dust samples were successfully predicted. ConclusionSignificant differences are observed in the fungal species composition and relative abundance among the cities of Beijing, Fuzhou, Kunming, and Urumqi. Employing fungal ITS2 sequencing on dust samples from these urban areas enables accurate inference of biogeographical locations. The high feasibility of utilizing fungal community data in dust for biogeographical inferences holds particular promise in the field of forensic science.

ObjectiveTo study the effects of applying different microbial fertilizers on the growth and rhizosphere soil environment of Codonopsis pilosula and provide a theoretical basis for ecological cultivation of this medicinal plant. MethodsSeven groups were designed， including CK （no application of microbial fertilizer）， T1 （Trichoderma longibrachiatum fertilizer）， T2 （Bacillus subtilis fertilizer）， T3 （Trichoderma viride fertilizer）， T4 （compound microbial fertilizer）， T5 （C. pilosula stems and leaves fermented with compound microbial fertilizer）， and T6 （Scutellaria baicalensis stems and leaves fermented with T. viride fertilizer）. The physiological indicators， yield， and quality of C. pilosula and the physicochemical properties， enzyme activities， and microbial diversity in the rhizosphere soil of different fertilizer treatments were measured. ResultsGroup T1 showed slight decreases in soluble protein content （SPC） and superoxide dismutase （SOD）. Groups T2-T6 showed increases in physiological indicators such as proline （Pro）， soluble solids content （SSC）， SPC， catalase （CAT）， and peroxidase （POD） and a decrease in malondialdehyde （MDA） in C. pilosula leaves. All the fertilizer treatments increased the yield of C. pilosula and the total polysaccharide content in the roots. T1， T2， T3， T4， and T5 increased the total flavonoid content in the roots. Meanwhile， T4 increased the total saponin content in the roots. All the fertilizer treatments reduced the pH and increased the electric conductivity （EC）， soil organic matter （SOM）， and alkaline nitrogen （AN） in the soil. T2 and T5 increased the available phosphorus （AP）， and T3， T4， T5， and T6 increased the available potassium （AK） in the soil. All the fertilizer treatments increased the activities of urease， sucrase， and CAT in the soil. Except that T1 decreased the bacterial diversity in the soil， other fertilizer treatments significantly increased bacterial and fungal diversity in the soil. Different fertilizer treatments significantly affected the composition of bacterial and fungal communities in the soil. At the phylum level， the dominant bacterial phyla included Proteobacteria， Acidobacteriota， and Bacteroideta， and the dominant fungal phyla were Ascomycota， Mortierellomycota， and unclassified_fungi in the rhizosphere soil of C. pilosula after bacterial fertilizer treatment. At the genus level， unclassified Gemmatimonadaceae， Sphingomonas， and unclassified Vicinamibacteraceae were the dominant bacterial genera， while unidentified， unclassified Fungi， and unclassified Sordariomycetes were the dominant fungal genera in the rhizosphere soil. The results of redundancy analysis indicated that the main physicochemical factors affecting changes of microbial communities in the rhizosphere soil of C. pilosula were pH， EC， AK， AN， AP， and soil organic matter （SOM） in the soil. The correlation heatmap showed that Bryobacter had significantly positive correlations with EC， AK， and AN. There was a significantly negative correlation between Fusarium and SOM. In summary， applying an appropriate amount of microbial fertilizer can promote the growth and improve the rhizosphere soil environment of C. pilosula. ConclusionThe compound microbial fertilizer and the C. pilosula stems and leaves fermented with compound microbial fertilizer can improve the soil nutrients， growth， development， yield， and quality of C. pilosula， and thus they can be applied to the artificial cultivation of C. pilosula.

ObjectiveTo study the effects of applying different microbial fertilizers on the growth and rhizosphere soil environment of Codonopsis pilosula and provide a theoretical basis for ecological cultivation of this medicinal plant. MethodsSeven groups were designed， including CK （no application of microbial fertilizer）， T1 （Trichoderma longibrachiatum fertilizer）， T2 （Bacillus subtilis fertilizer）， T3 （Trichoderma viride fertilizer）， T4 （compound microbial fertilizer）， T5 （C. pilosula stems and leaves fermented with compound microbial fertilizer）， and T6 （Scutellaria baicalensis stems and leaves fermented with T. viride fertilizer）. The physiological indicators， yield， and quality of C. pilosula and the physicochemical properties， enzyme activities， and microbial diversity in the rhizosphere soil of different fertilizer treatments were measured. ResultsGroup T1 showed slight decreases in soluble protein content （SPC） and superoxide dismutase （SOD）. Groups T2-T6 showed increases in physiological indicators such as proline （Pro）， soluble solids content （SSC）， SPC， catalase （CAT）， and peroxidase （POD） and a decrease in malondialdehyde （MDA） in C. pilosula leaves. All the fertilizer treatments increased the yield of C. pilosula and the total polysaccharide content in the roots. T1， T2， T3， T4， and T5 increased the total flavonoid content in the roots. Meanwhile， T4 increased the total saponin content in the roots. All the fertilizer treatments reduced the pH and increased the electric conductivity （EC）， soil organic matter （SOM）， and alkaline nitrogen （AN） in the soil. T2 and T5 increased the available phosphorus （AP）， and T3， T4， T5， and T6 increased the available potassium （AK） in the soil. All the fertilizer treatments increased the activities of urease， sucrase， and CAT in the soil. Except that T1 decreased the bacterial diversity in the soil， other fertilizer treatments significantly increased bacterial and fungal diversity in the soil. Different fertilizer treatments significantly affected the composition of bacterial and fungal communities in the soil. At the phylum level， the dominant bacterial phyla included Proteobacteria， Acidobacteriota， and Bacteroideta， and the dominant fungal phyla were Ascomycota， Mortierellomycota， and unclassified_fungi in the rhizosphere soil of C. pilosula after bacterial fertilizer treatment. At the genus level， unclassified Gemmatimonadaceae， Sphingomonas， and unclassified Vicinamibacteraceae were the dominant bacterial genera， while unidentified， unclassified Fungi， and unclassified Sordariomycetes were the dominant fungal genera in the rhizosphere soil. The results of redundancy analysis indicated that the main physicochemical factors affecting changes of microbial communities in the rhizosphere soil of C. pilosula were pH， EC， AK， AN， AP， and soil organic matter （SOM） in the soil. The correlation heatmap showed that Bryobacter had significantly positive correlations with EC， AK， and AN. There was a significantly negative correlation between Fusarium and SOM. In summary， applying an appropriate amount of microbial fertilizer can promote the growth and improve the rhizosphere soil environment of C. pilosula. ConclusionThe compound microbial fertilizer and the C. pilosula stems and leaves fermented with compound microbial fertilizer can improve the soil nutrients， growth， development， yield， and quality of C. pilosula， and thus they can be applied to the artificial cultivation of C. pilosula.

[摘 要] 目的：探讨程序性死亡蛋白1（PD-1）抑制剂与恶性黑色素瘤患者癌性疲劳（CRF）之间的关系及其影响因素。方法：选取2019年4月至2024年4月期间在济南市人民医院接受治疗的100例恶性黑色素瘤患者作为研究对象，使用中文版Piper疲劳量表评估这些患者在接受首次PD-1抑制剂治疗前后3个月内的疲劳状况。结果：使用PD-1抑制剂前后的患者疲劳评分存在显著差异（P < 0.001）。单变量分析发现，性别、吸烟史、肿瘤位置以及使用的PD-1抑制剂类型对于疲劳程度没有明显影响（均P > 0.05），而与年龄、疾病分期、贫血状态、白细胞减少症、继发性甲状腺机能减退（甲减）、继发性肾上腺皮质醇功能减退（AI）、继发性促肾上腺皮质激素降低（P < 0.05或P < 0.01或P < 0.001）相关联。进一步的多变量回归分析揭示，继发性甲减、继发性AI、贫血、白细胞减低是导致此类患者出现严重CRF的关键独立风险因素（均P < 0.05）。结论: PD-1抑制剂的不良反应继发性甲减、继发性AI、贫血、白细胞减低是恶性黑色素瘤患者CRF的独立风险因素。

ObjectiveMaxing Shigan Tang， as a traditional prescription for treating pneumonia， has a remarkable clinical effect. This study aims to systematically investigate the molecular mechanisms of Maxing Shigan Tang in treating pneumonia by integrating its structural and transcriptomic data at the target level. MethodsNP-TCMtarget， a developed systematic network pharmacological model focusing on drug targets， was used to mine the effect targets of Maxing Shigan Tang for treating pneumonia based on the transcriptome data. The structural targets of chemical components in Maxing Shigan Tang were predicted based on the structural information. The intersection of effect targets and structural targets was taken as the direct targets of Maxing Shigan Tang for treating pneumonia， and the remaining effect targets except direct targets were taken as indirect targets. Finally， functional enrichment analysis was performed on these targets to explore the molecular mechanism of Maxing Shigan Tang in treating pneumonia. ResultsA total of 1 604 effect targets and 816 structural targets of Maxing Shigan Tang for treating pneumonia were identified. Maxing Shigan Tang exerted its therapeutic effects through 164 direct targets and 1 440 indirect targets. The functional analysis of 1 604 effect targets predicted 19 significantly enriched pathways. Comprehensive analysis of these pathways showed that these targets were mainly linked to immune and inflammatory responses， such as cytokine-cytokine receptor interaction， necrosis factor （NF）-κB signaling pathway， and helper T cell 17 differentiation. ConclusionFocusing on the hierarchical feature of drug targets and the structural and transcriptomic data， this study systematically reveals the path of herbal component-direct target-indirect target-biological effects of Maxing Shigan Tang in treating pneumonia.

ObjectiveMaxing Shigan Tang， as a traditional prescription for treating pneumonia， has a remarkable clinical effect. This study aims to systematically investigate the molecular mechanisms of Maxing Shigan Tang in treating pneumonia by integrating its structural and transcriptomic data at the target level. MethodsNP-TCMtarget， a developed systematic network pharmacological model focusing on drug targets， was used to mine the effect targets of Maxing Shigan Tang for treating pneumonia based on the transcriptome data. The structural targets of chemical components in Maxing Shigan Tang were predicted based on the structural information. The intersection of effect targets and structural targets was taken as the direct targets of Maxing Shigan Tang for treating pneumonia， and the remaining effect targets except direct targets were taken as indirect targets. Finally， functional enrichment analysis was performed on these targets to explore the molecular mechanism of Maxing Shigan Tang in treating pneumonia. ResultsA total of 1 604 effect targets and 816 structural targets of Maxing Shigan Tang for treating pneumonia were identified. Maxing Shigan Tang exerted its therapeutic effects through 164 direct targets and 1 440 indirect targets. The functional analysis of 1 604 effect targets predicted 19 significantly enriched pathways. Comprehensive analysis of these pathways showed that these targets were mainly linked to immune and inflammatory responses， such as cytokine-cytokine receptor interaction， necrosis factor （NF）-κB signaling pathway， and helper T cell 17 differentiation. ConclusionFocusing on the hierarchical feature of drug targets and the structural and transcriptomic data， this study systematically reveals the path of herbal component-direct target-indirect target-biological effects of Maxing Shigan Tang in treating pneumonia.

OBJECTIVE To study the effects of peripheral blood-derived exosomes （Exo） intervened by Naozhenning （NZN） on injury of neuron cells HT22 induced by microglia BV-2 cells. METHODS Wistar rats were selected to prepare peripheral blood- derived Exo intervened by NZN （66.83 g/kg）， referred to as NZN-Exo； peripheral blood-derived Exo intervened by normal saline and piracetam （PLXT， 1.62 g/kg） were prepared using the same method， denoted as KB-Exo and PLXT-Exo respectively， and all Exo were subsequently identified. Meanwhile， BV-2 cells were stimulated with 1 μg/mL lipopolysaccharide （LPS） to prepare LPS- stimulated supernatant， and non-LPS-stimulated supernatant was prepared following the same protocol. HT22 cells were divided into four groups: KB-Exo group （treated with non-LPS-stimulated supernatant+KB-Exo）， model group （treated with LPS-stimulated supernatant+KB-Exo）， PLXT-Exo group （treated with LPS-stimulated supernatant+PLXT-Exo）， and NZN-Exo group （treated with LPS-stimulated supernatant+NZN-Exo）， with the concentration of the corresponding Exo in all groups being 50 μg/mL. After 24 hours of culture， the proliferation of HT22 cells was detected by the CCK-8 assay and EdU assay； the apoptosis of HT22 cells was detected； the microstructure of HT22 cells was observed； the contents of interleukin-1β （IL-1β）， IL-10， nuclear factor-κB （NF- κB）， and tumor necrosis factor-α （TNF-α） in HT22 cells were measured， as well as the expression levels of TNF-α， NOD-like receptor thermal protein domain associated protein 3 （NLRP3）， Caspase-1， B-cell lymphoma-2（ Bcl-2）， and Bcl-2-associated X protein （Bax）. RESULTS KB-Exo， PLXT-Exo and NZN-Exo were successfully prepared， and all Exo exhibited typical cup-shaped contours and membrane-enclosed characteristics. Compared with KB-Exo group， model group showed significantly decreased cell proliferation rates （detected by CCK-8 and EdU）， intracellular IL-10 levels， and Bcl-2 protein expression levels （P＜0.05）； while the cell apoptosis rate， intracellular levels of IL-1β， TNF-α， and NF-κB， as well as the expression levels of NLRP3， TNF-α， Caspase-1， and Bax proteins were significantly increased （P＜0.05）. Additionally， in the model group， the cells showed volume swelling， incomplete cell membrane， nucleolar rupture， significant swelling and deformation of mitochondria， and severe vacuolization. Compared with model group， the above quantitative indicators in the PLXT-Exo group and NZN-Exo group were significantly reversed （P＜0.05）， with large and round cell nuclei， intact nuclear membranes， and reduced mitochondrial vacuolization. CONCLUSIONS Peripheral blood-derived Exo intervened by naozhenning can alleviate the injury of neuronal cells HT22 by inhibiting inflammatory responses and cell apoptosis.