With the development of molecular pharmacognosy, the advantages of DNA molecular markers in the identification of original plants of Chinese medicinal materials are becoming increasingly significant. To compensate for the limitations of existing markers in the quality supervision of Chinese medicinal materials, our team has independently designed a new molecular marker named DNA signature sequence(DSS). This marker is a nucleotide sequence that only appears in a specific taxonomic unit, with a length of 40 bp and high identification accuracy. This article aims to screen and verify the DSS markers that can accurately identify the original plants of the medicinal materials included in the volume one of the Chinese Pharmacopoeia, establish the operating procedure for developing standard nucleotide sequences, and lay a foundation for the widespread application of polymerase chain reaction in the quality supervision of traditional Chinese medicine. Firstly, the Chloroplast Genome Information Resource(CGIR) was searched for the chloroplast genome sequences of the test samples, species of the same genus, and common background species. IdenDSS was used to obtain the DSS tags and specific identification primers of the tested species. After DNA extraction, PCR amplification, sequencing, and sequence alignments, a total of 203 DSS markers of Chinese medicinal materials were obtained for validation. The above sequences were uploaded to the Traditional Chinese Medicine Molecular Identification Platform(www.herbsdna.com), and a standard DSS library was established for identifying the original plants of medicinal materials, serving as an important tool for quality supervision of Chinese materia medica. On this basis, an operating procedure for DSS development is formed, laying a foundation for further DSS screening and application based on more diverse genome sequences.
Plants, Medicinal/classification* ; DNA, Plant/genetics* ; Drugs, Chinese Herbal ; Pharmacopoeias as Topic ; Genetic Markers ; Medicine, Chinese Traditional

AIM To study the secondary metabolites from endophytic fungi of Taxus wallichiana var.chinensis (Pilger) Florin and their biological activities.METHODS Endophytes were isolated and purified by tissue block and scribing method,the obtained strains were identified by molecular method.Macroporous resin column and silica gel column were used for separation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibition of phosphatidylinositol 3-kinase,phosphodiesterase and fusarium oxysporum activities were determined by chemiluminescence kinase assay kit method,resazurin cell viability assay and enzyme linked immunosorbent assay.RESULTS Eight endophytic fungi were isolated and identified as Trichoderma,Aspergillus,Gongronella,Phoma,Penicillium.Strain ZGG-5 had strong PI3K inhibitory activity with IC50 value of (0.17±0.05)μg/mL,and was identified as Gongronella butleri.Thirteen compounds were isolated from ZGG-5,and compound 13 had strong activities of inhibiting PI3K at concentration of 1 mg/mL,and the inhibitory rate was (97.8±0.51)％.CONCLUSION T.wallichiana var.chinensis contains endophytic fungi with strong activities of inhibiting PI3K.Compounds 1-12 are first isolated from G.butleri.

AIM To study the secondary metabolites from endophytic fungi of Taxus wallichiana var.chinensis (Pilger) Florin and their biological activities.METHODS Endophytes were isolated and purified by tissue block and scribing method,the obtained strains were identified by molecular method.Macroporous resin column and silica gel column were used for separation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibition of phosphatidylinositol 3-kinase,phosphodiesterase and fusarium oxysporum activities were determined by chemiluminescence kinase assay kit method,resazurin cell viability assay and enzyme linked immunosorbent assay.RESULTS Eight endophytic fungi were isolated and identified as Trichoderma,Aspergillus,Gongronella,Phoma,Penicillium.Strain ZGG-5 had strong PI3K inhibitory activity with IC50 value of (0.17±0.05)μg/mL,and was identified as Gongronella butleri.Thirteen compounds were isolated from ZGG-5,and compound 13 had strong activities of inhibiting PI3K at concentration of 1 mg/mL,and the inhibitory rate was (97.8±0.51)％.CONCLUSION T.wallichiana var.chinensis contains endophytic fungi with strong activities of inhibiting PI3K.Compounds 1-12 are first isolated from G.butleri.

There are inconsistencies in treatment outcomes, measurement instruments, and criteria for assessing clinical effectiveness in studies related to distal radius fractures (DRF), resulting in potential biases and failing to provide high-quality clinical evidence. To address these challenges, international researchers have reached a consensus on developing the core outcome indicator set for distal radius fractures(COS-DRF). However, it's important to note that the existing COS-DRF framework could not reflect the unique characteristics of Traditional Chinese Medicine (TCM) treatment. Currently, there are no established standards for treatment outcomes and measurement instruments specific to TCM clinical research, nor has a COS-DRF been established for TCM clinical studies in China. In light of these gaps, our research team aims to construct a core set of treatment outcomes for TCM clinical research on distal radius fractures. This involves compiling a comprehensive list of treatment outcomes and measurement instruments, initially derived from a thorough literature review and expert consensus, which will then undergo further refinement and updates based on real-world clinical experiences, incorporating feedback from 2 to 3 rounds of expert consensus or Delphi questionnaire surveys. Our goal is to establish a COS-DRF or CMS-DRF that aligns with the principles and practices of TCM, and provide high-quality evidence for clinical practice.
Humans ; China ; Consensus ; East Asian People ; Outcome Assessment, Health Care ; Wrist Fractures/therapy* ; Medicine, Chinese Traditional ; Review Literature as Topic ; Fracture Fixation/standards*

To explore the changes and the reaction mechanisms between soil microecological environment and the content of secon-dary metabolites of plants under water deficit, this study carried out a pot experiment on the 3-leaf stage seedlings of Rheum officinale to analyze their response mechanism under different drought gradients(normal water supply, mild, moderate, and severe drought). The results indicated that the content of flavonoids, phenols, terpenoids, and alkaloids in the root of R. officinale varied greatly under drought stresses. Under mild drought stress, the content of substances mentioned above was comparatively high, and the content of rutin, emodin, gallic acid, and(+)-catechin hydrate in the root significantly increased. The content of rutin, emodin, and gallic acid under severe drought stress was significantly lower than that under normal water supply. The number of species, Shannon diversity index, richness index, and Simpson index of bacteria in the rhizosphere soil were significantly higher than those in blank soil, and the number of microbial species and richness index decreased significantly with the aggravation of drought stresses. In the context of water deficit, Cyanophyta, Firmicutes, Actinobacteria, Chloroflexi, Gemmatimonadetes, Streptomyces, and Actinomyces were the dominant bacteria in the rhizosphere of R. officinale. The relative content of rutin and emodin in the root of R. officinale was positively correlated with the relative abundance of Cyanophyta and Firmicutes, and the relative content of(+)-catechin hydrate and(-)-epicatechin gallate was positively correlated with the relative abundance of Bacteroidetes and Firmicutes. In conclusion, appropriate drought stress can increase the content of secondary metabolites of R. officinale from physiological induction and the increase in the association with beneficial microbe.
Rhizosphere ; Rheum ; Droughts ; Soil ; Catechin ; Emodin ; Bacteria/metabolism* ; Water/metabolism* ; Firmicutes ; Soil Microbiology

Wound age estimation is the core content in the practice of forensic medicine. Accurate estimation of wound age is a scientific question that needs to be urgently solved by forensic scientists at home and abroad. Metabolomics techniques can effectively detect endogenous metabolites produced by internal or external stimulating factors and describe the dynamic changes of metabolites in vivo. It has the advantages of strong operability, high detection efficiency and accurate quantitative results. Machine learning algorithm has special advantages in processing high-dimensional data sets, which can effectively mine biological information and truly reflect the physiological, disease or injury state of the body. It is a new technical means for efficiently processing high-throughput big data. This paper reviews the status and advantages of metabolomic techniques combined with machine learning algorithm in the research of wound age estimation, and provides new ideas for this research.
Algorithms ; Machine Learning ; Forensic Medicine ; Metabolomics ; Big Data

ObjectiveTo explore the therapeutic effect and possible mechanism of Banxia Xiexintang and its disassembled prescriptions in regulating the flora disorder induced by mixed antibiotics in young rats. MethodSeventy male BALB/C young rats were randomly assigned into 7 groups： blank group， model group， Bifidobacterium tetralogy viable tablets （0.68 g·kg^-1） group， Banxia Xiexintang （9.1 g·kg^-1） group， Xinkai （3.19 g·kg^-1） group， Kujiang （1.82 g·kg^-1） group， and Ganbu （4.1 g·kg^-1） group， with 10 rats in each group. Except the blank group， the other groups were given mixed antibiotics by gavage to induce intestinal flora disorder. After 14 days， the rats in different drug groups were administrated with corresponding drugs by gavage， and those in the blank group and model group with the same amount of normal saline once a day for 14 days. After that， fecal samples were collected aseptically for 16S rDNA sequencing of intestinal flora， and lipopolysaccharide （LPS， 10 mg·kg^-1） was injected intraperitoneally to induce inflammatory reaction. The tissue morphology of colonic mucosa was observed via hematoxylin-eosin （HE） staining， and the macrophage infiltration of colonic mucosa was observed via toluidine blue staining and immunohistochemistry. The expression of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， tumor necrosis factor-α （TNF-α）， and interleukin-10 （IL-10） mRNA were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the modeling changed the intestinal flora structure of the young rats （P<0.01）， damaged the colonic mucosa， reduced the macrophage infiltration， and down-regulated the mRNA levels of IL-1β， IL-6， IL-8， TNF-α， and IL-10 （P<0.01）. Compared with the model group， bifidobacterium quadruple viable tablets， Banxia Xiexintang and its disassembled prescriptions increased the diversity of intestinal flora and the relative abundance of beneficial bacteria such as Bacteroidetes and Firmicutes （P<0.01）. At the same time， they ameliorated colonic mucosal injury （P<0.05， P<0.01）， increased macrophage infiltration （P<0.05， P<0.01）， and up-regulated the mRNA levels of IL-6， IL-8， and TNF-α （P<0.01）. The mRNA level of IL-1β was up-regulated in Bifidobacterium tetralogy viable tablets， Banxia Xiexintang， Kujiang， and Ganbu groups （P<0.01）， and that of IL-10 was up-regulated in Bifidobacterium tetralogy viable tablets， Banxia Xiexintang， Xinkai， and Ganbu groups （P<0.01）. ConclusionBanxia Xiexintang and the disassembled prescriptions can adjust the intestinal flora of young rats exposed to antibiotics and protect the immune barrier of colonic mucosa after intestinal flora disorder. In particularly， the whole prescription of Banxia Xiexintang demonstrates the best performance.

OBJECTIVES: The metabolomics technique of LC-MS/MS combined with data analysis was used to detect changes and differences in metabolic profiles in the vitreous humor of early rat carcasses found in water, and to explore the feasibility of its use for early postmortem submersion interval (PMSI) estimation and the cause of death determination. METHODS: The experimental model was established in natural lake water with 100 SD rats were randomly divided into a drowning group (n=50) and a postmortem (CO₂ suffocation) immediately submersion group (n=50). Vitreous humor was extracted from 10 rats in each group at 0, 6, 12, 18 and 24 h postmortem for metabolomics analyses, of which 8 were used as the training set to build the model, and 2 were used as test set. PCA and PLS multivariate statistical analysis were performed to explore the differences in metabolic profiles among PMSI and causes of death in the training set samples. Then random forest (RF) algorithm was used to screen several biomarkers to establish a model. RESULTS: PCA and PLS analysis showed that the metabolic profiles had time regularity, but no differences were found among different causes of death. Thirteen small molecule biomarkers with good temporal correlation were selected by RF algorithm. A simple PMSI estimation model was constructed based on this indicator set, and the data of the test samples showed the mean absolute error (MAE) of the model was 0.847 h. CONCLUSIONS The 13 metabolic markers screened in the vitreous humor of rat corpses in water had good correlations with the early PMSI. The simplified PMSI estimation model constructed by RF can be used to estimate the PMSI. Additionally, the metabolic profiles of vitreous humor cannot be used for early identification of cause of death in water carcasses.
Animals ; Biomarkers/metabolism* ; Cadaver ; Chromatography, Liquid ; Immersion ; Postmortem Changes ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Vitreous Body/metabolism* ; Water/metabolism*

Objective:To observe the clinical efficacy of balance-impact tuina therapy for lumbar intervertebral disc herniation (LIDH).Methods:A total of 118 eligible LIDH patients were randomized into an observation group and a control group by the random number table method,with 59 cases in each group.The observation group was intervened by balance-impact tuina therapy,while the control group was intervened by conventional tuina therapy,both for consecutive two weeks.The scores of visual analog scale (VAS),Oswestry disability index (ODI),quality of life questionnaire-core 30 (QOL-C30) were observed before and after treatment;the relapse rate was estimated at the sixth month and twelfth month following the treatment.The data were statistically analyzed.Results:After intervention,the total effective rate was 96.6％ in the observation group versus 91.5％ in the control group,and the between-group difference was statistically significant (P＜0.05).The VAS and ODI scores declined significantly after treatment in both groups (all P＜0.05),and the observation group was markedly lower than the control group (P＜0.05,P＜0.01).The QOL-C30 score increased significantly after treatment in both groups (both P＜0.05),and the observation group was markedly higher than the control group (P＜0.05).The relapse rates at the post-treatment sixth month and twelfth month in the observation group were lower than those in the control group (P＜0.05,P＜0.01).Conclusion:Compared with the conventional tuina therapy,the balance-impact tuina therapy shows advantage in lessening pain,improving the function and enhancing the quality of life in the treatment of LIDH,and it has a lower relapse rate.Thus,this therapy is worth promoting in clinic.

Objective To investigate the expression of cannabinoid type 2 receptor （CB2R） at different time points after brain contusion and its relationship with wound age of mice. Methods A mouse brain contusion model was established with PCI3000 Precision Cortical Impactor. Expression changes of CB2R around the injured area were detected with immunohistochemical staining, immunofluorescent staining and Western blotting at different time points. Results Immunohistochemical staining results showed that only a few cells in the cerebral cortex of the sham operated group had CB2R positive expression. The ratio of CB2R positive cells gradually increased after injury and reached the peak twice at 12 h and 7 d post-injury, followed by a decrease to the normal level 28 d post-injury. The results of Western blotting were consistent with the immunohistochemical staining results. Immunofluorescent staining demonstrated that the changes of the ratio of CB2R positive cells in neurons, CB2R positive cells in monocytes and CB2R positive cells in astrocytes to the total cell number showed a single peak pattern, which peaked at 12 h, 1 d and 7 d post-injury, respectively. Conclusion The expression of CB2R after brain contusion in neurons, monocytes and astrocytes in mice suggests that it is likely to be involved in the regulation of the biological functions of those cells. The changes in CB2R are time-dependent, which suggests its potential applicability as a biological indicator for wound age estimation of brain contusion in forensic practice.
Animals ; Blotting, Western ; Brain Contusion/metabolism* ; Brain Injuries ; Forensic Pathology ; Mice ; Muscle, Skeletal/pathology* ; Receptor, Cannabinoid, CB2/metabolism* ; Receptors, Cannabinoid ; Time Factors ; Wound Healing/physiology*