ObjectiveThis study aims to investigate the effects of different sizes of seedlings and melatonin treatment on physiological and biochemical indicators and bolting-related gene expression in Angelica sinensis， find substances related to early bolting， and elucidate the inhibitory mechanism of melatonin on bolting. MethodsSpectrophotometry was used to detect the related enzyme activities of A. sinensis leaves. The contents of endogenous hormones and polyamines were detected using ultra-high performance liquid chromatography-tandem mass spectrometry. Real-time polymerase chain reaction （Real-time PCR） was used to detect the expression levels of bolting-related genes. Inter-group differential indicator analysis， orthogonal partial least squares discriminant analysis， and principal component analysis were comprehensively applied to identify factors related to early bolting. ResultsEndogenous jasmonic acid and melatonin were identified as the most important factors affecting early bolting. Secondly， the activity of antioxidant enzymes， abscisic acid content， gibberellin content， and the expression levels of CO3， HD3A， and FD genes had important effects on the bolting process. Compared with small seedlings， exogenous melatonin treatment mainly inhibited early bolting by increasing endogenous melatonin content， reducing gibberellin content， and decreasing the expression levels of SOC1 and FD genes. ConclusionExogenous melatonin can inhibit early bolting in A. sinensis by regulating its physiological， biochemical， and gene expression levels.

ObjectiveThis study aims to investigate the effects of different sizes of seedlings and melatonin treatment on physiological and biochemical indicators and bolting-related gene expression in Angelica sinensis， find substances related to early bolting， and elucidate the inhibitory mechanism of melatonin on bolting. MethodsSpectrophotometry was used to detect the related enzyme activities of A. sinensis leaves. The contents of endogenous hormones and polyamines were detected using ultra-high performance liquid chromatography-tandem mass spectrometry. Real-time polymerase chain reaction （Real-time PCR） was used to detect the expression levels of bolting-related genes. Inter-group differential indicator analysis， orthogonal partial least squares discriminant analysis， and principal component analysis were comprehensively applied to identify factors related to early bolting. ResultsEndogenous jasmonic acid and melatonin were identified as the most important factors affecting early bolting. Secondly， the activity of antioxidant enzymes， abscisic acid content， gibberellin content， and the expression levels of CO3， HD3A， and FD genes had important effects on the bolting process. Compared with small seedlings， exogenous melatonin treatment mainly inhibited early bolting by increasing endogenous melatonin content， reducing gibberellin content， and decreasing the expression levels of SOC1 and FD genes. ConclusionExogenous melatonin can inhibit early bolting in A. sinensis by regulating its physiological， biochemical， and gene expression levels.

ObjectiveTo investigate the bacterial diversity in the rhizosphere soil and phyllosphere of Angelica sinensis and examine the effects of foliar applications of a composite bacterial agent，salicylic acid，and coronatine on the bacterial diversity，disease incidence，and plant yield，thus providing a theoretical basis and guidance for the artificial construction of functional minimal communities and the regulation of rhizosphere through foliar treatments. MethodsUnder continuous cropping conditions in the field，foliar applications of a composite bacterial agent，salicylic acid，coronatine，and sterile water were conducted. The 100-plant weight was measured via the conventional method，and the incidence of diseases was recorded. The microbial community composition，diversity，and inter-group differences in the phyllosphere and rhizosphere soil of A. sinensis were analyzed by 16S high-throughput sequencing，and the potential microbial functions were predicted. ResultsCompared with the blank control，foliar applications of salicylic acid and coronatine both significantly reduced the yield and root rot incidence of A. sinensis. The foliar application of salicylic acid decreased the content of ferulic acid and increased that of ligustilide. The foliar application of coronatine increased the content of both ferulic acid and ligustilide. The microbial communities and functions in the phyllosphere and rhizosphere soil were significantly different. The phyllosphere had lower microbial diversity，with all bacteria being Gram-negative，mainly Cyanobacteria and Proteobacteria with limited functions. The rhizosphere soil had higher microbial diversity，harboring dominant phyla including Proteobacteria，Actinobacteria，Acidobacteria，and Bacteroidetes with rich functions. All foliar treatments regulated the microbial community in the rhizosphere soil，with a more significant effect on the microbial community in the rhizosphere soil than that in the phyllosphere. The coronatine treatment significantly reduced the abundance of Proteobacteria and nitrate-reducing and aromatic compound-degrading microorganisms in the rhizosphere soil，thus affecting nutrient cycling and autotoxic substance degradation and leading to a yield reduction. Compared with the salicylic acid treatment，the coronatine treatment significantly increased the abundance of Bacillus and Streptomyces in the rhizosphere soil，demonstrating enhanced disease control efficacy. ConclusionFoliar application of coronatine and salicylic acid can significantly regulate the composition and function of bacterial communities in the rhizosphere soil，thereby reducing the disease incidence and the plant yield.

The aging trend is intensifying currently,but there is still a lack of standardized diagnosis and treat-ment schemes for severe infections in elderly people.This paper focuses on the recommendations for immune-related clinical diagnosis and treatment routes as well as the idea of risk stratified diagnosis and treatment for elderly peo-ple,aiming to effectively prevent infectious diseases in elderly people and perform stratified management through systematic and scientific means of immune function monitoring and regulation,so as to enhance the standardized level of diagnosis and treatment as well as clinical treatment effect of infection in elderly people.

The aging trend is intensifying currently,but there is still a lack of standardized diagnosis and treat-ment schemes for severe infections in elderly people.This paper focuses on the recommendations for immune-related clinical diagnosis and treatment routes as well as the idea of risk stratified diagnosis and treatment for elderly peo-ple,aiming to effectively prevent infectious diseases in elderly people and perform stratified management through systematic and scientific means of immune function monitoring and regulation,so as to enhance the standardized level of diagnosis and treatment as well as clinical treatment effect of infection in elderly people.

Evaluating the consistency of herb injectable formulations could improve their product quality and clinical safety, particularly concerning the composition and content levels of trace ingredients. Panax notoginseng Saponins Injection (PNSI), widely used in China for treating acute cardiovascular diseases, contains low-abundance (10%-25%) and trace saponins in addition to its five main constituents (notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Re, ginsenoside Rb₁, and ginsenoside Rd). This study aimed to establish a robust analytical method and assess the variability in trace saponin levels within PNSI from different vendors and formulation types. To achieve this, a liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) method employing multiple ions monitoring (MIM) was developed. A "post-column valve switching" strategy was implemented to eliminate highly abundant peaks (NR₁, Rg₁, and Re) at 26 min. A total of 51 saponins in PNSI were quantified or relatively quantified using 18 saponin standards, with digoxin as the internal standard. This study evaluated 119 batches of PNSI from seven vendors, revealing significant variability in trace saponin levels among different vendors and formulation types. These findings highlight the importance of consistent content in low-abundance and trace saponins to ensure product control and clinical safety. Standardization of these ingredients is crucial for maintaining the quality and effectiveness of PNSI in treating acute cardiovascular diseases.
Ginsenosides ; Saponins ; Chemometrics ; Panax notoginseng ; Cardiovascular Diseases ; Chromatography, Liquid ; Tandem Mass Spectrometry

Objective To review the efficacy and safety of combination of iodine-125 brachytherapy and concurrent chemoradiotherapy for middle and advanced stage non-small cell lung cancer (NSCLC).Methods Eighty-two cases of peripheral NSCLC patients were divided into the experimental group (n =32)and the control group (n =50) by different treatment methods.The patients in experimental group were treated by iodine-125 brachytherapy combined with concurrent chemoradiotherapy,and the patients in control group were treated by concurrent chemoradiotherapy.The two groups were treated with helical tomotherapy,and the chemotherapy regimen was PC regimen (paclitaxel + cisplatin) or TP regimen (taxotere + cisplatin).The response rate (RR),progression free survival (PFS),overall survival (OS) and adverse reactions were observed.Results The RR of experimental group and control group was 87.5％ and 76.0％,and the difference was not statistically significant (x2 =0.992,P =0.319).The median PFS was 16 months and 11 months,and the difference was statistically significant (x2 =5.216,P =0.022).The median OS was 26 months and 19 months,and the difference was not statistically significant (x2 =1.085,P =0.298).There were no statistically significant differences of the occurrence rates of adverse reactions in the experimental group and the control group such as radioactive pneumonia (18.75％ vs.24.00％,x2 =0.314,P =0.575),radiation esophagitis (25.00％ vs.30.00％,x2 =0.242,P =0.623),digestive tract reaction (21.88％ vs.26.00％,x2=0.180,P =0.671),bone marrow suppression (12.50％ vs.16.00％,x2 =0.014,P =0.907) and hemopneumothorax (6.25％ vs.0,P =0.149).Conclusion The combination of iodine-125 brachytherapy with concurrent chemoradiotherapy is a safe and effective treatment for middle and advanced stage NSCLC,which is worthy of further research and promotion.

Objective To manufacture magnetic microbubbles with dual-response to ultrasound and magnetic fields.Methods Microbubbles of ultrasound contrast agent (ST68) based on a surfactant were prepared by the acoustic cavitation method.Fe3O4 magnetic nanoparticles with negative charge were synthesized using the polyol procedure.Magnetic microbubbles were generated by depositing polyethylenimine and Fe3O4 magnetic nanoparticles alternately onto the microbubbles using the layer-by-layer self-assembly.In vitro ultrasonography was performed on a silicone tube with/without magnetic microbubbles (3 × 108/ml) by a self-made device to observe the movement of magnetic microbubbles under the effects of magnetic field.In vivo imaging was performed on the kidney of New Zealand rabbits before and after the injection of magnetic microbubbles.Results The Fe3O4 nanoparticles carried a stable negative charge of (-24.6 ± 6.7) mV and more than 98％ of the particles were less than 8 μm in diameter,meeting the size requirement of an ultrasound contrast agent for intravenous administration.There was no echoic signal in the silicone tube before injection of magnetic microbubbles,but there were strong echoic signals after injection.After applying a magnetic field,the magnetic microbubbles moved along the direction of the magnetic flux.In vivo ultrasound imaging could not visualize the kidney before injection of magnetic microbubbles,but could remarkably visualize the kidney after injection.Conclusions The magnetic microbubbles exhibit favorable magnetic targeting and ultrasound contrast enhancement characteristics.Such properties may serve as the foundation to study their potential for simultaneous diagnosis and treatment in the future.

Objective To fabricate an ultrasound/fluorescence bi-modal contrast agent by encapsulating fluorescent quantum dots into polymeric ultrasound contrast agent microbubbles.Methods Polylactic acid (PLA,500 mg),(1R)-(+)-camphor (50 mg) and CdSe/ZnS quantum dots (0.5 ml,2.3 μmol/L)were dissolved or dispersed in dichloromethane (10 ml) to form in an organic phase.Ammonium carbonate solution and poly (vinyl alcohol) solution were employed as the internal and external water phase,respectively.The fluorescent microbubbles were generated using double emulsion solvent evaporation and lyophilization methods.The morphology and illumination were characterized by scanning electron microscopy (SEM) and fluorescence spectrophotometry.Synchronized contrast-enhanced ultrasound and fluorescence imaging was acquired by injecting fluorescent microbubbles into the silicone tube coupled to a self-made ultrasound/fluorescence imaging device.Ultrasound/fluorescence bi-modal in vivo imaging was acquired on the kidney of New Zealand rabbits and suckling mice.Results The fluorescent microbubbles were hollow spheres with an averaged diameter of (1.62 ± 1.47) μm.More than 99％ of these microbubbles were less than 8 μm in diameter,which meeted the size criteria for ultrasound contrast agents.The fluorescence emission peak of the microbubbles appeared at 632 nm,indicating that good luminescence properties of quantum dots were maintained.In vitro ultrasound/fluorescence imaging showed no echoic signal when the silicone tube was filled with saline,but there was a strong echo when filled with fluorescent microbubbles.The liquid column with fluorescent microbubbles emitted red luminescence under ultraviolet irradiation.The kidney of the rabbit was remarkably enhanced after the administration of fluorescent microbubbles.Bright fluorescence could be observed at the injection site of the suckling mice via subcutaneous injection.Conclusions A bi-modal but single contrast agent based on polymeric microbubbles has been successfully fabricated for the use of ultrasound and fluorescence imaging.It retains the good characteristics of both echogenicity and fluorescence,which complement each other in case of limitations imposed by uni-modal,single agents.

Objective To study the regulation of heparanase expression by hypoxia and its correlation to the invasiveness of tumor cells. Methods BxPC-3 cells were cultured in hypoxia in vitro and the heparanase mRNA and protein expression were detected by reverse transcriptional polymerase reaction chains (RT-PCR) and western blot respectively. Matrigel invasion assay was used to observe the invasive abilities of tumor cells in hypoxia and in the status of heparanase was inhibited by antisense oligodeoxynucleotide (AS-ODN) targeting to the heparanase gene promoter. Results In normoxia, there was a relatively low level of heparanase mRNA and protein expression in cultured BxPC-3 cells. In hypoxia, heparanase expression, mRNA and protein which expressed consistently, were inhibited slightly at 3h and upregulated significantly at 6h, 12h, 24h and 48h. When the heparanase expression was inhibited by AS-ODN, the heparanase mRNA and protein maintained low in hypoxia, however, the nonsense oligodeoxynucleotide (NS-ODN) did not block upregulation of heparanase expression. In matrigel assay, after 48h incubation, number of BxPC-3 cells that penetrated the Matrigel-coated filter of transwell chamber was increased 96.2% in hypoxia (P<0.01), the Hpa AS-ODN (400 nmol/L) inhibited the invasive cells by 37.2% (P<0.05). Conclusions BxPC-3 cells invasion ability is enhanced by hypoxia through upregulation of heparanase mRNA and protein expression in BxPC-3 pancreatic cancer cell lines.