ObjectiveTo investigate the effects of rhizosphere organic acids secreted by the roots of Salvia miltiorrhiza on continuous cropping obstacles. MethodsThe mixed solution of organic acids in the rhizosphere of S. miltiorrhiza in continuous cropping and rotation cropping was added to the hairy roots subcultured for 21 days， and samples were collected on days 0， 2， 4， 6， 8， and 10. The changes of biomass， effective components， primary metabolites， secondary metabolites， antioxidant enzymes， and hormones in hairy roots of S. miltiorrhiza were observed and determined. ResultsCompared with the rotation cropping group and the blank control group， the simulation of organic acid secretion from the roots of S. miltiorrhiza had a significant inhibitory effect on the growth of hairy roots and decreased the content of effective components as well as total sugar and total protein in primary metabolites. Compared with the blank control group， the rotation cropping group and the continuous cropping group showed total sugar and total protein content decreases of 33.9% and 5.1%， respectively. On the other hand， the secretion of organic acids from S. miltiorrhiza roots significantly promoted the accumulation of total phenolic acids and total tanshinone， which showed increases of 14.6% and 1.6%， respectively， in continuous cropping group and rotation cropping group compared with the blank control group. ConclusionThe organic acid environment under continuous cropping significantly inhibited the growth of hairy roots and the accumulation of primary metabolites， while promoting the synthesis and accumulation of secondary metabolites of S. miltiorrhiza.

Alternative splicing (AS) serves as a fundamental regulatory mechanism in gene expression, contributing to proteomic diversity by generating an array of mRNA isoforms from precursor mRNA via distinct splice site combinations. In light of the limited therapeutic options currently available, the exploration of AS as a target for drug development is of paramount importance. This review offers an exhaustive analysis of the biological functions and underlying molecular mechanisms associated with various AS-induced splice variants, RNA-binding proteins, and cis-elements, highlighting their significance as clinical biomarkers. We place particular emphasis on the current therapeutic applications of AS in an array of lung diseases, including but not limited to lung cancer, cystic fibrosis, silicosis, acute respiratory distress syndrome, pneumonia, asthma, chronic obstructive pulmonary diseases, pulmonary arterial hypertension, and idiopathic pulmonary fibrosis. The review delves into the role of AS events in the diagnosis and treatment of lung diseases, focusing on the regulatory influence of splicing factors and RNA-binding proteins, while also enumerating the mutated components implicated in AS misregulation. Consequently, a comprehensive understanding of the intricate mechanisms governing these splicing events could potentially offer novel avenues for the development of splicing-targeted therapeutics and diagnostic tools for the prevention and treatment of lung diseases.

ObjectiveTo explore the effects of angle and original moisture content on the moisture distribution， migration and contents of effective components in the drying process of sliced Salviae Miltiorrhizae Radix et Rhizoma（SMRR）. MethodsSet the slicing angles of SMRR at 30°， 45°， and 90°. Cut the fresh samples， 1/3 dehydrated samples， and 2/3 dehydrated samples， dry them in an oven at 40 ℃ and take samples at the set time points. Low-field nuclear magnetic resonance（LF-NMR） and magnetic resonance imaging（MRI） were used to analyze the changes in transverse relaxation time（T₂） of SMRR samples in 9 treatment groups at specific times， as well as the distribution and migration of water in the samples. The contents of tanshinone Ⅱ_A， tanshinone Ⅰ， cryptotanshinone， and salvianolic acid B in samples from 9 different treatment groups were determined by high performance liquid chromatography（HPLC）， and the best processing technology of SMRR was screened by combining with One-way ANOVA， Duncan multiple comparison and principal component analysis（PCA）. ResultsThe moisture content of dry basis of SMRR in each treatment group decreased with the extension of drying time. The drying rate of fresh cut group decreased slowly at first， while the drying rate of water loss group showed a trend of increasing at first and then decreasing. The internal water of SMRR could be divided into three states， including bound water， non flowing water and free water. During the drying process， the water migration law showed that the free water of fresh cut group disappeared after drying for 12 h， the content of bound water gradually decreased， and the overall fluidity deteriorated. In the water loss group， part of the free water was transformed into more cohesive and non flowing water after drying for 3 h， and the three kinds of water basically disappeared after drying for 12 h. The MRI results showed that the entire dehydration process slowly moved from the outer side to the center， and the internal water eventually dissipated. In terms of the contents of active ingredients， the order of the effect of slicing angle on the total content of active ingredients in SMRR was 30°>45°>90°. The content of tanshinones was ranked as 1/3 dehydrated group>2/3 dehydrated group>fresh cut group， and the content of salvianolic acid B was ranked as 1/3 dehydrated group>fresh cut group>2/3 dehydrated group. Combined with the results of PCA and comprehensive scoring results， the overall level of effective component content in SMRR was the highest when cut at 30° after 1/3 of water loss. ConclusionAfter comprehensive evaluation， SMRR can be sliced at 30° after 1/3 of water loss. It is not only easy to cut， but also the surface and cross-sectional colors remain basically unchanged after drying， which is similar to the color under traditional processing， and the effective ingredients are preserved the highest. This study can provide a basis for the optimization of processing technology of SMRR.

Diabetic retinopathy (DR) is a common complication of diabetes that can lead to visual impairment or even blindness. Current treatments mainly rely on invasive methods, which carry the risk of complications, making early intervention crucial. In recent years, research has revealed that the mechanical microenvironment of the retina plays a key role in the development and progression of DR, involving cell migration, functional disorders, and changes in the extracellular matrix. Mechanobiological mechanisms, such as mechanical signal transduction and the Hippo/Yes-associated protein pathway, are gradually being uncovered. Mechanical diagnostic and therapeutic strategies, including optical coherence elastography, tissue engineering, and intelligent diagnostic systems, offer new directions for DR management. In the future, further integration of biomechanics and mechanobiology research is needed to build multi-scale mechanical models and explore the synergistic regulation mechanisms between mechanical and biochemical factors, aiming to achieve precise and personalized diagnosis and treatment of DR and ultimately improve patients' visual outcomes.

Diabetic retinopathy (DR) is a common complication of diabetes that can lead to visual impairment or even blindness. Current treatments mainly rely on invasive methods, which carry the risk of complications, making early intervention crucial. In recent years, research has revealed that the mechanical microenvironment of the retina plays a key role in the development and progression of DR, involving cell migration, functional disorders, and changes in the extracellular matrix. Mechanobiological mechanisms, such as mechanical signal transduction and the Hippo/Yes-associated protein pathway, are gradually being uncovered. Mechanical diagnostic and therapeutic strategies, including optical coherence elastography, tissue engineering, and intelligent diagnostic systems, offer new directions for DR management. In the future, further integration of biomechanics and mechanobiology research is needed to build multi-scale mechanical models and explore the synergistic regulation mechanisms between mechanical and biochemical factors, aiming to achieve precise and personalized diagnosis and treatment of DR and ultimately improve patients' visual outcomes.

Objective:To observe the long-term efficacy and safety of macular buckling (MB) in the treatment of high myopia traction maculopathy.Methods:A retrospective clinical study. From January 2014 to December 2017, 57 eyes of 57 patients with high myopia traction maculopathy who underwent MB treatment at Zhongshan Ophthalmic Center of Sun Yat-sen University were included in the study. Among them, there were 15 males with 15 eyes, average age was 51.80±10.72 years; there were 42 females with 42 eyes, average age was 59.14±11.51 years. There were 21 eyes of 21 cases with highly myopic macular hole with macular detachment (MHMD), and 36 eyes in 36 cases with highly myopic foveoschisis with macular detachment (FSMD), and they were grouped accordingly. All patients underwent best corrected visual acuity (BCVA), optical coherence tomography (OCT), and axial length (AL) measurements. The standard logarithmic visual acuity chart was used for BCVA examination, which was converted into logarithm of the minimum angle of resolution (logMAR) visual acuity during statistics. All patients underwent MB, either on its own or combined with vitrectomy. Patients with significant vitreous macular traction on OCT were treated with combined surgery. One, 3, 6 months and 1, 2, 3, and 4 years after the operation, the same equipment and methods before the operation were used to conduct related examinations, and the long-term efficacy and safety of the two groups of eyes were observed.Results:Before surgery, the logMAR BCVA of eyes in MHMD group and FSMD group were 1.35±0.47 and 1.17±0.59, respectively; 4 years after surgery, they were 1.02±0.49 and 0.73±0.55, respectively. The BCVA improved significantly at postoperative 4 years than preoperative in both groups ( P=0.039, 0.001). In the eyes with MHMD, the BCVA was found to be significant improved 3 years after surgery ( P=0.042). Whereas, in the eyes with FSMD, the BCVA was found to be significantly improved 3 months after surgery ( P=0.013). Macular reattachment was achieved in 100% of cases, while macular hole closure rate was achieved in 66.7% in the MHMD group. In the FSMD group, either macular reattachment rate or the foveoschisis resolution rate was 97.2%. After surgery, choroidal neovascularization was observed in 2 eyes, and 3 eyes with intraretinal cyst. Conclusion:MB may represent a safe and effective surgical option for the treatment of high myopia maculopathy.

Salvia miltiorrhiza Bge(SMB)has long been used in traditional Chinese medicine to treat cardiovascular and cerebrovascular diseases.Growing clinical usage has led to a huge demand for artificial planting of SMB.Thus,continuous cropping of SMB is an important challenge that needs to be addressed.Contin-uous cropping can alter the metabolic profile of plants,resulting in poor growth and low yield.In this study,we tried to image the spatial location and variation of endogenous metabolites in continuously cropped SMB using matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI).Spatially resolved expressions of tanshinones,salvianolic acids,polyamines,phenolic acids,amino acids,and oligosaccharides in normal and continuously cropped SMB roots were compared.The ex-pressions of dihydrotanshinone Ⅰ,tanshinone Ⅱ A,dehydromiltirone,miltirone,dehydrotanshinone ⅡA,spermine,salvianolic acid B/E,tetrasaccharide,and pentasaccharide in continuously cropped SMB roots were much lower than those in normal roots.There was little difference in the expressions of caffeic acid and salvianolic acid A in normal and continuously cropped SMB roots.Ferulic acid was more widely distributed in xylem of normal SMB but strongly expressed in xylem,phloem,and cambium of continuously cropped SMB.The spatially resolved metabolite information enhances our understanding of the metabolic signature of continuously cropped SMB and also provides insights into the metabolic ef-fects of continuous cropping in other plants.

Objective: To study the effect of WT1 expression on the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in acute leukemia (AL) and its significance as molecular marker to dynamically monitor minimal residual disease (MRD) . Methods: Retrospectively analyzed those AL patients who underwent allo-HSCT in the First Hospital Affiliated to Zhejiang University School of Medicine during Jan 2016 to Dec 2017, a total number of 314 cases, 163 males and 151 females, median age was 30 (9-64) years old. Comparing the difference of WT1 expression at diagnosed, pre-HSCT and after HSCT. Using the receiver operating characteristic (ROC) curve to determine the WT1 threshold at different time so as to predict relapse. The threshold of WT1 expression before transplantation was 1.010%, within 3 months after HSCT was 0.079% and 6 months after HSCT was 0.375%. According to these thresholds, WT1 positive patients were divided into low expression groups and high expression groups. Analyzed the relationship between overall survival (OS) , disease-free survival (DFS) , cumulative incidence of relapse (CIR) and WT1 expression. Results: The OS and DFS of high expression group pre-HSCT were lower than low expression group [69.2% (9/13) vs 89.1% (57/64) , χ²=4.086, P=0.043; 53.8% (7/13) vs 87.5% (56/64) , χ²=9.766, P=0.002], CIR was higher than low expression group [30.8% (4/13) vs 7.8% (5/64) , P=0.017]. There was no significant difference of OS and DFS between high expression and low expression group of 3 months after HSCT (P=0.558, P=0.269) . The OS and DFS of high expression group of 6 months after transplantation were both lower than low expression group (P=0.049, P=0.035) . Multivariate analysis showed that WT1>0.375% when 6 months after transplantation was the only independent prognostic factor for shorter DFS (P=0.022) . There was no statistically significant difference in CIR between the high-expression group and the low-expression group 3 months after transplantation and 6 months after transplantation (P=0.114, P=0.306) . Conclusion High expression of WT1 before and after HSCT was an adverse prognosis factor. It is of clinical practical value to use WT1 as a transplant recommendation index for patients with acute leukemia and as a marker to monitor MRD dynamically.

BACKGROUND:Chitosan biological materials can induce bone marrow mesenchymal stem cells to differentiate toward neurons. As a derivative of chitosan, carboxymethyl chitosan has a series of excelent properties. However, whether carboxymethyl chitosan can induce the neuronal differentiation of bone marrow mesenchymal stem cells remains unclear.OBJECTIVE:To investigate the effect of carboxymethyl chitosan thermosensitive hydrogel on the differentiation of bone marrow mesenchymal stem cells into neurons and the possible mechanism.METHODS:Passage 3 bone marrow mesenchymal stem cells from rats were selected and cultured in carboxymethyl chitosan thermosensitive hydrogel extracts in different concentrations (0, 50, 100, 150, 200, 500 g/L). Control cells were cultured in culture medium with no addition of carboxymethyl chitosan thermosensitive hydrogel extracts. MTT assay was performed to investigate the effects of different concentrations of carboxymethyl chitosan thermosensitive hydrogel extracts on bone marrow mesenchymal stem cell proliferation. Western blot assay was used to explore the effect of 150 g/L carboxymethyl chitosan thermosensitive hydrogel extracts on the expression of neuron-specific enolase, Nestin, Vimentin, NF-M, microtubule associated protein 2, glial fibrillary acidic protein, β3-tubulin, Notch1 and Jag1 protein.RESULTS AND CONCLUSION:MTT assay showed that carboxymethyl chitosan thermosensitive hydrogel promoted the cell proliferation, and the proliferation rate reached the peak at the concentration of 150 g/L. Western blot assay showed that the cells induced by 150 g/L carboxymethyl chitosan thermosensitive hydrogel extract had significant increases in neuron-specific enolase, Nestin, Vimentin, NF-M, microtubule associated protein 2, glial fibrillary acidic protein, and β3-tubulin protein expression, and obvious decreases in Notch1 and Jag1 protein expression in comparison with the control group. These results indicate that the carboxymethyl chitosan thermosensitive hydrogel induces rat bone marrow mesenchymal stem cells to differentiate toward neurons, and suppresses the activity of Notch signal pathway in the process of differentiation.

Objective To investigate the condition of flight fatigue and its relationship with mental health in the pilots of base transformation on the purpose of providing the reference for comprehensive intervention during the transformation.Methods Both 90 transformed and 90 nontransformed pilots were tested by fatigue assessment instrument (FAI) and symptom checklist 90 (SCL 90).Thirty-eight transformed and 38 non-transformed pilots were selected to continuously record ECG for 1 h for the analysis of heart rate variability (HRV).Correlation analysis was done on the condition of fatigue and mental health.Results ①FAI score results:the total score and factor 1,2,and 3 of FAI in transformed pilots was significantly higher than that in non-transformed pilots (t=1.981-4.570,P＜0.05).②HRV test:the standard deviation of R-R intervals,root mean square of the successive R R intervals difference,total power,low frequency,high frequency in transformed pilots was all significantly lower than that in non-transformed pilots (t=4.475-8.644,P＜0.01).The ratio of low frequency to high frequency of transformed pilots was significantly higher than that of non-transformed pilots (t=7.681,P＜0.01).③The total score and the score of factor 1,2,and 3 in FAI showed varied positive correlation with the total score and each score of factors in SCL-90 (r=0.165-0.452,P＜0.01 or 0.05).The score of factor 4 in FAI had weak positive correlation with the total score of SCL-90 and the score of sensitivity in interpersonal relationship,depression,hostility (r=0.158-0.175,P＜0.05).Conclusions The severity of flight fatigue in transformed pilots was significantly higher than that in non transformed pilots.The condition of flight fatigue is closely correlated with mental health.We should carry out comprehensive intervention based on psychological intervention to ensure the flight safety during transformation.