OBJECTIVE To prepare reactive oxygen species （ROS）-responsive methotrexate （MTX）-modified paclitaxel （PTX）/icariin （ICA） micelles （MTX-oxi-Ms@PTX/ICA）， and perform technology optimization and in vitro anti-tumor effect evaluation. METHODS Synergistic toxicity concentration range of PTX and ICA was screened by synergistic toxicity test. The micelles were prepared by thin film hydration method， and their technology was optimized by response surface methodology. The fundamental characteristics of the micelles prepared by the optimal technology were evaluated. The micelles’ cytotoxicity， targeting ability to renal carcinoma RENCA cells of mice， and their inhibitory effects on invasion and migration were assessed. RESULTS Results of synergistic toxicity experiments demonstrated that the strongest synergistic effect occurred when PTX concentrations ranged from 2.5 to 10 μmol/L and ICA concentrations ranged from 5 to 15 μmol/L. The optimal technology of MTX-oxi-Ms@PTX/ ICA was determined to include 80 mg Soluplus®， Soluplus® and TPGS1000 mass ratio of 4∶1 （mg/mg）， 2 mg DSPE-PEG2000-TK- PEG5000， 2 mg DSPE-PEG2000-MTX， 1 mg PTX， and 1.5 mg ICA， with a hydration temperature of 35 ℃ and a formulation volume of 5 mL. Under the optimal conditions， average encapsulation efficiency of PTX and ICA in 3 batches of MTX-oxi- Ms@PTX/ICA reached 92.75%， the critical micelle concentration （CMC） was 0.007 9 mg/mL， the particle size was （62.09±1.68） nm， the polydispersity index （PDI） was 0.046±0.032， and the Zeta potential was （－2.47±0.15） mV. Within 30 days of placement， there was no significant change E-mail：yingqiang_1126@163.com in particle size and polydispersity index of micelle. In vitro release experiments showed that MTX-oxi-Ms@PTX/ICA released drugs more rapidly in oxidative environments. The half maximal inhibitory concentration of MTX-oxi-Ms@PTX/ICA against RENCA cells was （5.170±0.036） μmol/L. In vitro cellular uptake experiments indicated that compared with unmodified micelles， MTX modified micelles had stronger targeting effects on cancer cells， and also significantly enhanced the inhibitory ability of invasion and migration of RENCA cells （P＜0.05）. CONCLUSIONS MTX-oxi-Ms@PTX/ICA micelles are successfully prepared， which exhibit high encapsulation efficiency， low critical micelle concentration， and good stability. These micelles demonstrate significant cytotoxicity against RENCA cells and effectively inhibit cancer cell invasion and migration.

Mitochondrial dysfunction in chondrocytes is a key pathogenic factor in osteoarthritis (OA), but directly modulating mitochondria in vivo remains a significant challenge. This study is the first to verify a correlation between mitochondrial dysfunction and the downregulation of the FOXO3 gene in the cartilage of OA patients, highlighting the potential for regulating mitophagy via FOXO3 gene modulation to alleviate OA. Consequently, we developed a chondrocyte-targeting CRISPR/Cas9-based FOXO3 gene-editing tool (FoxO3) and integrated it within a nanoengineered 'truck' (NETT, FoxO3-NETT). This was further encapsulated in injectable hydrogel microspheres (FoxO3-NETT@SMs) to harness the antioxidant properties of sodium alginate and the enhanced lubrication of hybrid exosomes. Collectively, these FoxO3-NETT@SMs successfully activate mitophagy and rebalance mitochondrial function in OA chondrocytes through the Foxo3 gene-modulated PINK1/Parkin pathway. As a result, FoxO3-NETT@SMs stimulate chondrocytes proliferation, migration, and ECM production in vitro, and effectively alleviate OA progression in vivo, demonstrating significant potential for clinical applications.

Sesquiterpenes are natural terpenes containing 15 carbon atoms. They are widely used in the perfume, pharmaceutical, and biofuel industries due to their remarkable biological activities. The traditional production of sesquiterpenes relies on chemical synthesis or plant extraction, which has the disadvantages of low yields and waste of resources. The construction of microbial cell factories for the efficient synthesis of sesquiterpenes by means of synthetic biology provides a new option. In recent years, with the development of metabolic engineering and synthetic biology, the heterologous synthesis of a variety of sesquiterpenes has been successfully achieved by metabolic engineering of the oleaginous yeast, Yarrowia lipolytica. In this paper, we review the research progress in the heterologous synthesis of different sesquiterpenes by Y. lipolytica, discuss the synthetic biology strategies commonly used in this field, and make an outlook on the research directions and engineering approaches to further enhance the sesquiterpene yield in this host. This paper provides a reference for strategies such as synergistic optimization of synthetic biology and metabolic engineering, enhanced precursors, and opens up new directions for the application of synthetic biology in green chemistry and sustainable production.
Yarrowia/genetics* ; Sesquiterpenes/metabolism* ; Metabolic Engineering/methods* ; Synthetic Biology/methods*

Objective:To observe the application value of "Internet +" based case teaching method combined with objective structured clinical examination (OSCE) teaching method in the teaching of nursing students in the Department of Nephrology, and analyze its impact on nursing student satisfaction of teaching.Methods:A total of 60 nursing students who practiced in the Department of Nephrology, Jiangsu Provincial People's Hospital from January 2021 to June 2022 were selected as the research subjects. They were randomly divided into observation group and control group, with 30 students in each group. The control group adopted traditional teaching method, and the observation group adopted case teaching method based on "Internet +" combined with OSCE teaching method. SPSS 25.0 statistical software was used for data analysis, and independent sample t-test was used to compare the clinical practical work ability and teaching satisfaction of the two groups of nursing students after teaching. Results:After 4 weeks of teaching, the total scores of clinical thinking ability, disease observation and evaluation ability, mastery of medical knowledge, standardized operation ability, communicative ability, adaptive ability, humanistic care and health literacy of the observation group were higher than those of the control group [(29.84±3.52) vs. (25.67±3.36), (5.81±1.52) vs. (4.34±1.67), (17.93±2.24) vs. (15.52±2.05), (25.72±2.73) vs. (22.17±2.56), (4.18±0.75) vs. (3.56±0.69), (83.48±6.67) vs. (71.26±7.05)], with a statistically significant difference ( P < 0.05); after 4 weeks of teaching, the teaching preparation, teaching process, teaching ability, teaching attitude, teaching effect and total score of the observation group were higher than those of the control group [(17.13±2.42) vs. (15.07±1.84), (36.44±3.12) vs. (34.07±2.49), (21.97±2.36) vs. (21.37±2.01),(17.00±2.36) vs. (16.83±2.23), (17.73±2.00) vs. (14.07±2.55), (110.27±5.51) vs. (101.40±4.58)], and the difference was statistically significant ( P<0.05). Conclusions:The case teaching method based on "Internet +" combined with OSCE can not only effectively improve the clinical practical work ability of nursing students in the department of nephrology, but also improve student satisfaction of teaching.

Objective:To evaluate the efficacy and prognostic factors of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with secondary acute myeloid leukemia (sAML) .Methods:In this multicenter, retrospective clinical study, adult patients aged ≥18 years who underwent allo-HSCT for sAML at four centers of the Zhejiang Hematopoietic Stem Cell Transplantation Collaborative Group from January 2014 to November 2022 were included, and the efficacy and prognostic factors of allo-HSCT were analyzed.Results:A total of 95 patients were enrolled; 66 (69.5%) had myelodysplastic syndrome-acute myeloid leukemia (MDS-AML) , 4 (4.2%) had MDS/MPN-AML, and 25 (26.3%) had therapy-related AML (tAML) . The 3-year CIR, LFS, and overall survival (OS) rates were 18.6% (95% CI 10.2%-27.0%) , 70.6% (95% CI 60.8%-80.4%) , and 73.3% (95% CI 63.9%-82.7%) , respectively. The 3-year CIRs of the M-AML group (including MDS-AML and MDS/MPN-AML) and the tAML group were 20.0% and 16.4%, respectively ( P=0.430) . The 3-year LFSs were 68.3% and 75.4%, respectively ( P=0.176) . The 3-year OS rates were 69.7% and 75.4%, respectively ( P=0.233) . The 3-year CIRs of the groups with and without TP53 mutations were 60.0% and 13.7%, respectively ( P=0.003) ; the 3-year LFSs were 20.0% and 76.5%, respectively ( P=0.002) ; and the 3-year OS rates were 40.0% and 77.6%, respectively ( P=0.002) . According to European LeukmiaNet 2022 (ELN2022) risk stratification, the 3-year CIRs of patients in the low-, intermediate-, and high-risk groups were 8.3%, 17.8%, and 22.6%, respectively ( P=0.639) . The three-year LFSs were 91.7%, 69.5%, and 65.6%, respectively ( P=0.268) . The 3-year OS rates were 91.7%, 71.4%, and 70.1%, respectively ( P=0.314) . Multivariate analysis revealed that advanced disease at allo-HSCT and TP53 mutations were independent risk factors for CIR, LFS, and OS. Conclusion:There was no significant difference in the prognosis of patients who underwent allo-HSCT among the MDS-AML, MDS/MPN-AML, and tAML groups. Advanced disease at transplantation and TP53 mutations were poor prognostic factors. ELN2022 risk stratification had limited value for predicting the prognosis of patients with sAML following allo-HSCT.

Objective To explore the regulatory and functional mechanism of angiopoietin-like 4(ANGPTL4)expression in gastric epithelial cells infected with Helicobacter pylori(H.pylori).Methods H.pylori-positive gastric specimens from Department of Gastroenterology of second Affiliated Hospital of Army Medical University in November 2021 and the gastric tissues of H.pylori-infected mice were collected,and the expression locations of ANGPTL4 in these gastric tissues were detected by immunofluorescence assay.After gastric epithelial cells and gastric organoids were infected with H.pylori,the expression of ANGPTL4 was detected by real-time PCR and Western blotting.The regulatory mechanism of H.pylori-induced ANGPTL4 expression was investigated by cellular models above.Recombinant ANGPTL4 was used to stimulate gastric epithelial cells to investigate the effect of ANGPTL4 on the claudin 1(CLDN1)expression.Results H.pylori infection significantly increased the expression of ANGPTL4 in gastric epithelial cells in a CagA-dependent,bacterial concentration-dependent and infection time-dependent manner(P＜0.05).Compared to uninfected cells and CagA knockout(ΔcagA)H.pylori-induced ones,the ANGPTL4 expression was significantly increased in WT H.pylori-infected human and mouse gastric organoid(P＜0.05).Blocking NF-κB signaling pathway significantly inhibited the enhanced ANGPTL4 expression in H.pylori-infected gastric epithelial cells(P＜0.05).Recombinant ANGPTL4 activated ERK signaling pathway to down-regulate the CLDN1 expression in gastric epithelial cells(P＜0.05).Conclusion H.pylori activates NF-κB signaling pathway to up-regulate ANGPTL4 expression in gastric epithelial cells in a CagA-dependent manner,and ANGPTL4 activates ERK signaling pathway to down-regulate the CLDN1 expression in gastric epithelial cells,which may contributes to the development of gastric diseases induced by H.pylori infection.

Objective:To establish the standard of nursing quality in bone oncology department, and provide the basis for scientific evaluation of nursing quality in bone oncology department.Methods:On the theoretical basis of Donabedian′s three-dimensional quality model of "structure-process-outcome", and through literature review and semi-structured interview method, the "evaluation index of nursing quality in bone oncology department" was preliminatively formulated from November 2022 to June 2023. The Delphi method was used to select 31 experts from 31 third-level A hospitals and nursing colleges in 27 provinces or municipalities across the country for two rounds of correspondence consultation. The criteria were screened and modified to determine the evaluation criteria of nursing quality in bone tumor specialty.Results:The questionnaire recovery rate of 2 rounds of expert consultation was 100.00%, the authority coefficient of 2 rounds of expert consultation was 0.93, and the coefficient of variation of 1, 2 and 3-grade standards were all less than or equal to 0.25. The Kendall′s coefficient of concordance of the primary standards of the two rounds of expert consultation were both 0.088, in the secondary standards were 0.103 and 0.140, in the tertiary standards were 0.119 and 0.110. Through 2 rounds of expert letter consultation, the evaluation criteria for the quality of care in bone tumor specialties were divided into three levels, including 3 primary standards (structural quality criteria, process quality criteria and outcome quality criteria), 21 secondary standards and 80 tertiary standards.Conclusions:The construction process of nursing quality standard in bone oncology department is scientific and reliable, reflecting specialty characteristics, and can provide scientific basis for the evaluation of nursing quality in bone oncology department and standardize nursing behavior.

Seeds are the source for the production of Chinese medicinal materials. The seed authenticity and quality of directly affect the effectiveness and safety of Chinese medicinal materials. The seed quality is faced with the problems such as mixed sources， existence of adulterants and seeds stocked for years， low maturity， and low purity. To ensure the high-quality and sustainable development of the Chinese medicinal material industry， it is urgent to standardize the seed market and identify and evaluate the quality of the seeds circulating in the market. Seed identification methods include visual inspection， microscopic observation， micro-character identification， chemical fingerprinting， molecular identification， electronic nose， X-ray diffraction， electrochemical fingerprinting， spectral imaging， and artificial intelligence. These methods have different application scopes and unique advantages and disadvantages. According to the different species of Chinese herbal medicines and different requirements of testing sites， suitable methods can be selected to achieve rapid and accurate identification with low costs. In the future， the seed identification methods should be developed based on emerging technologies with interdisciplinary knowledge， and intelligent， nondestructive， and single-grain detection methods are needed for the modern Chinese medicinal material industry. This paper introduces the seed identification technologies currently applied in research and production， compares the principles， applicability， advantages， and disadvantages of different technologies， and provides an outlook on the future development of seed identification technologies， aiming to provide a reference for the identification and quality evaluation of seeds of Chinese medicinal material.

ObjectiveTo establish a method based on specific polymerase chain reaction （PCR） that can accurately and rapidly identify Astragalus membranaceus var. mongholicus （AMM） seeds and A. membranaceus （AM） seeds. MethodThe Chloroplast Genome Information Resource （CGIR） and IdenDSS were used to obtain the characteristic DNA fragments of AMM and AM， and the specific single nucleotide polymorphism （SNP） sites of AMM and AM were screened out， on the basis of which the specific primers MG-F/MG-R of AMM and MJ-F/MJ-R of AM were designed. The specific PCR method for identifying AMM and AM was established and optimized， and the specificity and applicability of the method were investigated. ResultThe specific PCR conditions for the identification of AMM were primers MG-F/MG-R， annealing at 62 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the specific band appeared at about 220 bp， with no band for the seeds of AM or adulterants. The specific PCR conditions for identifying the AM were primers MJ-F/MJ-R， annealing at 58 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the band appeared at about 150 bp， with no band of AMM or adulterants. ConclusionThe specific PCR method established in this study can accurately and quickly identify the seeds of AMM and AM， which provides a basis for the classification and accurate identification of Astragalus seeds and adulterants.

BACKGROUND:At present,wrist protection products designed in and outside China have not solved the contradiction between protecting the wrist joint from injury and maintaining the flexible movement of the wrist joint. OBJECTIVE:To investigate the biomechanical mechanism of dorsiflexion injury of the wrist joint in elderly people after falls,and to provide a biomechanical basis for the prevention and treatment of wrist injury in elderly people after falls. METHODS:A 65-year-old man was selected to obtain the original data by uninterrupted CT scan of the middle and lower 2/3 of his left forearm up to the end of the finger.A finite element model of wrist dorsiflexion was established using ANSYS 12.0 finite element software.The palm surface of the model was constrained,and the model at a velocity load of 2 m/s in the direction of vertical downward was given to simulate the injury state of the palm when the elderly fall.The stress distribution of the soft tissues and bones of the wrist joint and the change of the stress with time were observed after the load was applied. RESULTS AND CONCLUSION:(1)A realistic and effective finite element model of the dorsal extension position of the wrist joint of the elderly was established.The soft tissue stresses were mainly concentrated in the small fissure of the palm and the dorsal side of the wrist after loading.The skeletal stresses were mainly concentrated in the lower end of the ulnar radius dorsally.The stresses in the lower end of the radius were the greatest.The palmar stresses were mainly concentrated in the middle and lower 1/3 of the radius and the hook bone.The stress distribution of the ulnar radius was asymmetric,and the stresses in the radius were more concentrated.(2)The results of the study are consistent with the clinical situation of a fallen wrist injury in elderly people,and can be used to explain the mechanism of wrist dorsiflexion injury,which can provide the biomechanical basis for the design of wrist protection devices that can be used to prevent wrist injury induced by falling and the treatment of wrist injury in elderly people.