The bioactivity-guided isolation of potentially active natural products has been widely utilized in pharmaceutical discovery. In this study, by screening fungal extracts against coxsackievirus B3 (CVB3), three new aspochalasins, templichalasins A‒C (1‒3), along with six known aspochalasins (4‒9) were isolated from an active extract derived from the endophytic fungus Aspergillus templicola LHWf045. Compound 1 features a unique 5/6/5/7/5 pentacyclic ring system, while compounds 2 and 3 possess unusual 5/6/6/7 tetracyclic skeletons. Their structures were characterized through extensive spectroscopic analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. Additionally, we demonstrated that compound 4 can be readily converted into compounds 1‒3 under mild acidic conditions and proposed a plausible mechanism for this conversion. Bioactivity evaluation of compounds 1‒9 against CVB3 revealed the inhibitory effects of all compounds against the virus. Notably, compound 9 exhibited superior antiviral activity, surpassing the commercial drug ribavirin in selectivity index (SI) value.
Antiviral Agents/isolation & purification* ; Aspergillus/chemistry* ; Molecular Structure ; Enterovirus B, Human/drug effects* ; Endophytes/chemistry* ; Cytochalasins/isolation & purification* ; Drug Discovery ; Humans

This study aims to integrate data mining techniques of single cell transcriptomics and spatial transcriptomics, along with animal experiment validation, so as to systematically characterize the protective effects of Jianpi Tongluo Formula(JTF) on the cartilage in knee osteoarthritis(KOA) and elucidate the underlying molecular mechanisms. Single cell transcriptomics and spatial transcriptomics datasets(GSE254844 and GSE255460) of the cartilage tissue obtained from KOA patients were analyzed to map the single cell-spatial heterogeneity and identify key pathogenic factors. After that, a KOA rat model was established via knee joint injection of papain. The intervention effects of JTF on the expression features of these key factors were assessed through real-time quantitative polymerase chain reaction(PCR), Western blot, and immunohistochemical staining. As a result, the integrated single cell and spatial transcriptomics data identified distinct cell subsets with different pathological changes in different regions of the inflamed cartilage tissue in KOA, and their differentiation trajectories were closely related to the inflammatory fibrosis-like pathological changes of chondrocytes. Accordingly, the expression levels of the two key effect targets, namely nuclear receptor coactivator 4(NCOA4) and high mobility group box 1(HMGB1) were significantly reduced in the articular surface and superficial zone of the inflamed joints when JTF effectively alleviated various pathological changes in KOA rats, thus reversing the abnormal chondrocyte autophagy level, relieving the inflammatory responses and fibrosis-like pathological changes, and promoting the repair of chondrocyte function. Collectively, this study revealed the heterogeneous characteristics and dynamic changes of inflamed cartilage tissue in different regions and different cell subsets in KOA patients. It is worth noting that NCOA4 and HMGB1 were crucial in regulating chondrocyte autophagy and inflammatory reaction, while JTF could reverse the regulation of NCOA4 and HMGB1 and correct the abnormal molecular signal axis in the target cells of the inflamed joints. The research can provide a new research idea and scientific basis for developing a personalized therapeutic schedule targeting the spatiotemporal heterogeneity characteristics of KOA.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Rats ; Osteoarthritis, Knee/pathology* ; Humans ; Male ; Cartilage, Articular/metabolism* ; Chondrocytes/metabolism* ; Rats, Sprague-Dawley ; Female ; Protective Agents/administration & dosage* ; Single-Cell Analysis ; Middle Aged ; HMGB1 Protein/metabolism*

A pharmacophore-based study was conducted to investigate the therapeutic activity of the traditional Tibetan medicine Zha Xun (ZX) in liver diseases. In the present study, the protective effect of ZX on the acute liver injury induced by concanavalin A (ConA) and 0.15% carbon tetrachloride (0.15% CCl₄) in ICR mice was evaluated, and the results showed that ZX significantly reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the ConA-induced acute immune liver injury model and the CCl₄-induced acute oxidative liver injury model (P < 0.05). Subsequently, the protective effects of aqueous, 95% ethanol, 60% ethanol and 30% ethanol eluting fractions of ZX, and fulvic acid, the main water-soluble constituent of ZX, were evaluated against acute oxidative liver injury induced by 0.15% CCl₄ in mice. The results showed that different solvent-eluting fractions of ZX showed certain hepatoprotective activities, among which the aqueous extract of ZX and 30% ethanol extract of ZX significantly reduced the serum levels of ALT, AST, and lactate dehydrogenase (LDH) in mice (P < 0.05), and the serum levels of LDH in mice were significantly reduced by fulvic acid (P < 0.05), which showed significant hepatoprotective activity. The protective activities and preliminary mechanisms of the total extract of ZX, the aqueous extract of ZX, the 30% ethanol extract of ZX, and fulvic acid against hepatocellular injury in vitro were further evaluated by using the H₂O₂-induced hepatocellular injury model. The results showed that the components could significantly inhibit H₂O₂-induced hepatocellular injury, reduce the levels of ALT, alkaline phosphatase (ALP), and LDH, improve the survival rate of hepatocellular cells, and reduce the content of intracellular reactive oxygen species (ROS) in cell culture. At the same time, it can inhibit hepatocyte apoptosis by increasing the expression ratio of Bcl-2/BAX protein and decreasing the expression ratio of cleaved caspase-3/pro caspase-3 protein. The present study showed that ZX has clear hepatoprotective activity in vitro and in vivo, and the different solvent elution fractions of ZX showed certain hepatoprotective activity, among which the aqueous extract of ZX, 30% ethanol extract of ZX had better hepatoprotective activity, and the activity of 60% ethanol extract of ZX was stronger than that of 95% ethanol extract of ZX. The activity of ZX and its water-soluble elution site exerted hepatoprotective effects by inhibiting hepatocyte apoptosis and oxidative stress. The animals used in this experiment and related disposal meet the requirements of animal welfare, and have been reviewed and approved by the Laboratory Animal Management and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences (approval number: 00004018).

The study aimed to explore the effects of inoculation of Rhizophagus intraradices on the biomass, effective component content, and endogenous hormone content of Salvia miltiorrhiza through pot experiments. The number of leaves, plant height, dry weight of aboveground and underground parts, branch number, root number, root length, root diameter, and other biomass were mea-sured by weighing and counting methods. The content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, tanshinone Ⅱ_A, cryptotanshinone, and other effective components was determined by ultra-high performance liquid chromatography. The content of ABA and GA_3 was determined by triple quadrupole mass spectrometry. The correlations between biomass and effective components and between effective components and plant hormones ABA and GA_3 were analyzed. The results showed that R. intraradices significan-tly increased the aboveground dry weight, leaf number, and root number of S. miltiorrhiza by 0.24-0.65 times, respectively. The content of salvianolic acid B and rosmarinic acid in the aboveground part and the content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, and tanshinone Ⅱ_A in the underground part were significantly increased by 0.44-1.78 times, respectively. R. intraradices infection significantly increased the GA_3/ABA values of aboveground and underground parts by 3.82 and 76.47 times, respectively. The correlation analysis showed that caffeic acid, the effective component of the aboveground part, was significantly positively correlated with plant height, tanshinone Ⅱ_A, the effective component of the underground part, was significantly positively correlated with biomass root number, cryptotanshinone, and dry weight, while rosmarinic acid was significantly negatively correlated with dry weight. There were significant positive correlations between cryptotanshinone and ABA, tanshinone Ⅱ_A and ABA and GA_3, and caffeic acid and GA_3. In conclusion, R. intraradices can promote the accumulation of biomass and secondary metabolites of S. miltiorrhiza and regulate the balance between plant hormones ABA and GA_3, thereby promoting the growth of S. miltiorrhiza.
Salvia miltiorrhiza/chemistry* ; Plant Growth Regulators/analysis* ; Plant Roots/chemistry*

Objective:To compare the cumulative live birth rate (CLBR) per oocyte retrieval cycle between gonadotropin-releasing hormone (GnRH) antagonist protocol and progestin-primed ovarian stimulation (PPOS) protocol in patients aged 20-50 years.Methods:A retrospective cohort study was conducted to analyze 3 752 infertile patients aged 20-50 years who received in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). They used either GnRH antagonist protocol or PPOS protocol at the Center of Assisted Reproduction in Shanghai First Maternity and Infant Hospital from January 2017 to April 2021. One to one propensity score matching (PSM) was used to match the population characteristics. Baseline, clinical and laboratory characteristics, as well as pregnancy outcomes were compared between the two groups. The differences of CLBR was analyzed by multivariate logistic regression and subgroup analysis. Results:After matching, 1 466 patients (733 in each group) were included in the analysis. No significant differences were detected in age, body mass index, infertility type, cause and duration of infertility, number of stimulation cycles, basal follicle-stimulating hormone, number of antral follicles and composition ratio of insemination methods between the two groups ( P>0.05). Serum estradiol level [1 700.30 (1 011.76, 2 580.50) ng/L] and luteinizing hormone (LH) level [1.95 (1.07, 5.27) U/L] on trigger day were significantly lower in GnRH antagonist group than in PPOS group [2 056.50 (884.08, 3 601.59) ng/L, P=0.010; 3.00 (1.51, 5.00) U/L, P<0.001]. The cycle cancellation rate of PPOS group [30.56% (224/733)] was significantly higher than that of GnRH antagonist group [18.83% (138/733), P<0.001]. The numbers of oocytes obtained, available embryos and good-quality embryos were similar to those in GnRH antagonist group (all P>0.05). For each embryo transfer cycle, the implantation rate [16.97% (207/1 220) vs. 21.42% (266/1 242)], the clinical pregnancy rate [21.78% (188/863) vs. 27.38% (233/851)], the onging pregnancy rate [16.11% (139/863) vs. 21.62% (184/851)] and the live birth rate [15.06% (130/863) vs. 20.80% (177/851)] were significantly lower in PPOS group than in GnRH antagonist group ( P=0.010, P=0.012, P=0.004 and P=0.002, respectively). The CLBR of PPOS group was significantly lower than that of GnRH antagonist group [17.74% (130/733) vs. 24.15% (177/733), P=0.003]. Multivariate logistic regression analysis showed that ovarian stimulation protocol was an independent risk factor for CLBR [ OR=1.42, 95% CI: 1.03-1.95, P=0.032]. The results of subgroup analysis showed that the CLBR of PPOS group was significantly lower than that of GnRH antagonist group in the population aged ≤35 years and underwent non-first IVF/ICSI cycle [21.35% (111/520) vs. 28.93% (151/522), P=0.005; 7.85% (41/522) vs. 12.23% (62/507), P=0.019]. Conclusion:Compared with PPOS regimen, antagonist regimen can improve the CLBR per oocyte cycle in infertile patients aged 20-50 years, and is more significant in women aged ≤35 years and non-first oocyte collection patients.

Objective:To compare the cumulative live birth rate (CLBR) per oocyte retrieval cycle between gonadotropin-releasing hormone (GnRH) antagonist protocol and progestin-primed ovarian stimulation (PPOS) protocol in patients aged 20-50 years.Methods:A retrospective cohort study was conducted to analyze 3 752 infertile patients aged 20-50 years who received in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). They used either GnRH antagonist protocol or PPOS protocol at the Center of Assisted Reproduction in Shanghai First Maternity and Infant Hospital from January 2017 to April 2021. One to one propensity score matching (PSM) was used to match the population characteristics. Baseline, clinical and laboratory characteristics, as well as pregnancy outcomes were compared between the two groups. The differences of CLBR was analyzed by multivariate logistic regression and subgroup analysis. Results:After matching, 1 466 patients (733 in each group) were included in the analysis. No significant differences were detected in age, body mass index, infertility type, cause and duration of infertility, number of stimulation cycles, basal follicle-stimulating hormone, number of antral follicles and composition ratio of insemination methods between the two groups ( P>0.05). Serum estradiol level [1 700.30 (1 011.76, 2 580.50) ng/L] and luteinizing hormone (LH) level [1.95 (1.07, 5.27) U/L] on trigger day were significantly lower in GnRH antagonist group than in PPOS group [2 056.50 (884.08, 3 601.59) ng/L, P=0.010; 3.00 (1.51, 5.00) U/L, P<0.001]. The cycle cancellation rate of PPOS group [30.56% (224/733)] was significantly higher than that of GnRH antagonist group [18.83% (138/733), P<0.001]. The numbers of oocytes obtained, available embryos and good-quality embryos were similar to those in GnRH antagonist group (all P>0.05). For each embryo transfer cycle, the implantation rate [16.97% (207/1 220) vs. 21.42% (266/1 242)], the clinical pregnancy rate [21.78% (188/863) vs. 27.38% (233/851)], the onging pregnancy rate [16.11% (139/863) vs. 21.62% (184/851)] and the live birth rate [15.06% (130/863) vs. 20.80% (177/851)] were significantly lower in PPOS group than in GnRH antagonist group ( P=0.010, P=0.012, P=0.004 and P=0.002, respectively). The CLBR of PPOS group was significantly lower than that of GnRH antagonist group [17.74% (130/733) vs. 24.15% (177/733), P=0.003]. Multivariate logistic regression analysis showed that ovarian stimulation protocol was an independent risk factor for CLBR [ OR=1.42, 95% CI: 1.03-1.95, P=0.032]. The results of subgroup analysis showed that the CLBR of PPOS group was significantly lower than that of GnRH antagonist group in the population aged ≤35 years and underwent non-first IVF/ICSI cycle [21.35% (111/520) vs. 28.93% (151/522), P=0.005; 7.85% (41/522) vs. 12.23% (62/507), P=0.019]. Conclusion:Compared with PPOS regimen, antagonist regimen can improve the CLBR per oocyte cycle in infertile patients aged 20-50 years, and is more significant in women aged ≤35 years and non-first oocyte collection patients.

ObjectiveTo explore the radiosensitization and underlying mechanism of Xuefu Zhuyutang on subcutaneous transplanted esophageal carcinoma. MethodThe subcutaneous xenograft model of human esophageal carcinoma ECA-109 in nude mice was induced and the model mice were divided into a model group， an irradiation group， a Xuefu Zhuyutang group， and a combination group， with six nude mice in each group. After the intervention， the transplanted tumors were removed and weighed， and the tumor inhibition rate of each group was calculated according to the formula. The protein expression of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor A （VEGFA） was detected by immunohistochemistry （IHC）. The protein expression of mammalian target of rapamycin （mTOR）， HIF-1α， VEGFA， and vascular endothelial growth factor receptor 2 （VEGFR2） in transplanted tumors was detected by Western blot. The mRNA expression of mTOR， HIF-1α， and VEGFA in transplanted tumors was detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the conditions in the model group， the tumor weight decreased in the irradiation group and the Xuefu Zhuyutang group （P<0.05）， as well as the combination group （P<0.01）. Compared with the irradiation group， the combination group showed decreased tumor weight （P<0.05）， with tumor inhibition rate of 57.37%. Compared with the model group， the irradiation group， the Xuefu Zhuyutang group， and the combination group showed decreased protein expression of VEGFR2， p-mTOR， HIF-1α， and VEGFA （P<0.05， P<0.01） and reduced mRNA expression of mTOR， HIF-1α， and VEGFA （P<0.05， P<0.01）. Compared with the irradiation group， the combination group showed down-regulated protein expression of VEGFR2， p-mTOR， HIF-1α， and VEGFA （P<0.05， P<0.01） and reduced mRNA expression of mTOR， HIF-1α， and VEGFA （P<0.05， P<0.01）. ConclusionXuefu Zhuyutang can inhibit the growth of transplanted esophageal carcinoma ECA-109 in nude mice and shows an obvious radiosensitization effect in combination with radiotherapy. The mechanism may be related to the inhibition of the mTOR/HIF-1α/VEGFA signaling pathway to improve the hypoxic state of tumors.

OBJECTIVE: To establish the fixation model of anterior cervical transpedicular system (ACTPS) after subtotal resection of two segments of lower cervical spine(C₃-C₇) in order to provide a finite element modeling method for anterior cervical reconstruction. METHODS: The CT data of the cervical segment (C₁-T₁) of a 30-year-old adult healthy male volunteer was collected. Used Mimics 10.0, Rapidform XOR3, HyperMesh 10.0, CATIA5V19 and ANSYS 14.0 to establish the three-dimensional nonlinear complete model of lower cervical spine(C₃-C₇) as the intact group. The number of units and nodes of the complete model were recorded. After the effectiveness of the complete model was verified, the C₅ and C₆ vertebral subtotal resection was performed, and the ACTPS model was established as the ACTPS group. The axial force of 75 N and moment couple of 1N·m was loaded on the upper surface of C₃ in intact group and ACTPS group, the range of motion(ROM)and stress distribution in states of flexion extension, lateral flexion, rotation was compared between two groups. RESULTS: There were 85 832 elements and 23 612 nodes in the complete model of lower cervical spine(C₃-C₇) which was established in this experiment. The stress distribution of ACTPS internal fixation model was relatively uniform. Comparing with the intact group, the overall range of motion in ACTPS group was decreased in flexion extension, lateral flexion and rotation directions, and the corresponding compensation of adjacent C_3,4 segment was increased slightly. CONCLUSION The stress distribution of ACTPS fixation system is uniform, there is no stress concentration area at the joint of screw and titanium plate, and the fracture risk of internal fixation is low. It is suitable for stability reconstruction after anterior decompression of two or more cervical segments.
Adult ; Biomechanical Phenomena ; Bone Screws ; Cervical Vertebrae/surgery* ; Finite Element Analysis ; Humans ; Male ; Range of Motion, Articular ; Spinal Fusion

Arbuscular mycorrhizal fungi provided is beneficial to Salvia miltiorrhiza for increasing yield, promoting the accumulation of active ingredients, and alleviating S. miltiorrhiza disease etc. However, the application of fungicides will affect the benefit of arbuscular mycorrhizal fungi and there is little research about it. This article study the effect of four different fungicides: carbendazim, polyoxin, methyl mopazine, and mancozeb on mycorrhiza benefit to S. miltiorrhiza by the infection intensity of arbuscular mycorrhizal fungi, the growth of S. miltiorrhiza, and the content of active ingredients. RESULTS:: showed that different fungicides had different effects. The application of mancozeb had the strongest inhibitory effect on the mycorrhizal benefit to S. miltiorrhiza. Mancozeb significantly reduced the mycorrhizal colonization and the beneficial effect of arbuscular mycorrhizal fungi on the growth and the accumulation of active components of S. miltiorrhiza. The application of polyoxin had no significant effect on mycorrhizal colonization. Instead, it had a synergistic effect with the mycorrhizal benefit to promoting the growth and accumulation of rosmarinic acid of S. miltiorrhiza. The inhibitory strengths of four fungicides are: mancozeb>thiophanate methyl, carbendazim>polyoxin. Therefore, we recommend applying biological fungicides polyoxin and avoid applying chemical fungicides mancozeb for disease control during mycorrhizal cultivation of S. miltiorrhiza.
Fungicides, Industrial/pharmacology* ; Mycorrhizae ; Plant Roots ; Salvia miltiorrhiza ; Symbiosis

OBJECTIVE: To compare accuracy of anterior cervical pedicle screws between assist of rapid prototyping 3D guide plate and free-hand insertion, and evaluate the safety of two methods. METHODS: Eight adult cervical cadaver specimens after formaldehyde immersion, including 4 males and 4 females, aged 32 to 65(40.3±5.6) years old. After X-ray examination to exclude bone damage and deformity, 4 of them (3D guide plate group) randomly selected were for CT scan to obtain DICOM format data, and the data was imported into Mimics software for model, designed the ideal entry point and nail path for anterior cervicaltranspedicular screw (ATPS). After obtaining the personalized guide plate of the nail channel, it was exported as STL data, and the individual guide plate was printed by rapid prototyping and 3D printing technology. In turn, with the assistance of 3D guide plates, one-to-one personalized ATPS screws were placed on the four lower cervical cadaver specimens. Another 4 (free-hand group) lower cervical cadaver specimens were implanted with ATPS screws using free-hand technique. All specimens were performed CT thin-layer scanning and three-dimensional reconstruction after operation. The Tomasino method was used to evaluate the safety of the screws on the CT cross-sectional and sagittal images, to determine whether there was a cortical puncture of the lower and inner edges of the pedicle. According to the CT rating results, gradeⅠandⅡwere safe, and grade Ⅲ- Ⅴ were dangerous.And the accuracy of screws was recorded and analyzed between two groups. RESULTS: Two screws were inserted in each segment from C CONCLUSION The 3D printing rapid prototyping guide plate assisted insertion of the anterior cervical pedicle screw can significantly improve the accuracy and safety, and provide a theoretical basis for further clinical application.
Adult ; Aged ; Bone Plates ; Cervical Vertebrae/surgery* ; Cross-Sectional Studies ; Female ; Humans ; Male ; Middle Aged ; Pedicle Screws ; Printing, Three-Dimensional