ObjectiveTo explore the changes in color， odor and chemical components during wine-processing of Chuanxiong Rhizoma（CR）， identify differential markers， and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis（PLS-DA）， principal component analysis（PCA）， discriminant factor analysis（DFA） and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models， and differential components were screened out based on variable importance in the projection（VIP） value1. Then， high performance liquid chromatography（HPLC） was employed to detect the content changes of four components（ferulic acid， senkyunolide I， senkyunolide A and ligustilide） during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L^*（brightness value）， a^*（red-green value） and b^*（yellow-blue value）. Based on chromaticity differences， the color reference ranges were established for moderately processed CR， including L^* of 46.75-48.24， a^* of 5.37-6.07 and b^* of 20.32-21.70. In odor analysis， DFA revealed significant differences among processing stages， and 11 odor markers were identified， with four differential markers（4-hydroxy-3-butylphthalide， isopropyl butyrate， L-limonene and 1-methoxyhexane） based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection， with discrimination accuracy rates of 92.4% and 93.272% for color and odor， respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages， and four differential components were screened out， providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.

ObjectiveTo explore the changes in color， odor and chemical components during wine-processing of Chuanxiong Rhizoma（CR）， identify differential markers， and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis（PLS-DA）， principal component analysis（PCA）， discriminant factor analysis（DFA） and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models， and differential components were screened out based on variable importance in the projection（VIP） value1. Then， high performance liquid chromatography（HPLC） was employed to detect the content changes of four components（ferulic acid， senkyunolide I， senkyunolide A and ligustilide） during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L^*（brightness value）， a^*（red-green value） and b^*（yellow-blue value）. Based on chromaticity differences， the color reference ranges were established for moderately processed CR， including L^* of 46.75-48.24， a^* of 5.37-6.07 and b^* of 20.32-21.70. In odor analysis， DFA revealed significant differences among processing stages， and 11 odor markers were identified， with four differential markers（4-hydroxy-3-butylphthalide， isopropyl butyrate， L-limonene and 1-methoxyhexane） based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection， with discrimination accuracy rates of 92.4% and 93.272% for color and odor， respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages， and four differential components were screened out， providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.

Idiopathic pulmonary fibrosis （IPF） can be classified as pulmonary collateral disease，and its pathogenesis is mainly characterized by the loss of Qi meridian nourishment，the loss of Yin meridian nourishment，and the formation of blood stasis in the blood vessels. Qi Yin deficiency is the pathological basis that runs through IPF，and obstruction of meridians and collaterals is a key element in the development of the disease. The dysfunction of "spleen being in charge of the defensive function" is closely related to the formation of the pathological pattern of "lung deficiency and collateral stasis" in IPF. The term "spleen being in charge of the defensive function" originated from the Yellow Emperor's Inner Canon. If the spleen is healthy，the Qi will be filled with vitality. Positive energy is stored inside，evil cannot be dried up. Its concept is quite similar to the immune defense function in modern medicine. If the principle of "spleen being in charge of the defensive function" is lost，the key structure and function of the IPF alveolar epithelial barrier may be abnormal，and it can interact with various innate immune cells to promote inflammation and fibrosis processes. Therefore，this article explains the imbalance of immune homeostasis in IPF alveolar epithelium from two aspects：the barrier function of alveolar epithelial cells（AECs） and their interaction with innate immune cells. And based on the theory of "spleen being in charge of the defensive function"，using traditional Chinese medicine for strengthening the spleen and nourishing Qi to treat IPF from the perspective of the spleen. This not only strengthens the scientific connotation of "spleen being in charge of the defensive function" in the pathogenesis of IPF，but also provides new research directions and ideas for its future clinical prevention and treatment.

Objective To establish and verify a universal and stable potency test method in vitro for SARS-CoV-2 mRNA vaccine,so as to use it for the quality control of SARS-CoV-2 mRNA vaccine.Methods ELISA kits that could bind well to S protein of SARS-CoV-2 variants,as well as transfected cells,cell plating concentrations and doses for transfection were screened,and then a potency test method for SARS-CoV-2 mRNA vaccine in vitro was established and verified.Results An ELISA kit was found with good binding ability to S protein of each variant,and HEK293T cells were determined as the transfection cells,with the plating concentration of 2.5 × 10~5 cells/mL and the transfection dose of 4 μg/well in the 6-well plate.An universal and stable potency test method for SARS-CoV-2 mRNA vaccine in vitro was established.The verification results showed that the method met the quality control needs.Conclusion The established potency test method in vitro for SARS-CoV-2 mRNA vaccine has good relative accuracy,linearity,intermediate precision and range,and can be applied to the quality control of SARS-CoV-2 mRNA vaccines.

BACKGROUND:Conductive biomaterials are considered potential candidates for transmitting electrical signals for myocardial repair.Combining cell-based or cell-free strategies with conductive biomaterials to replenish cardiomyocytes and/or restore electrical signaling pathways is a promising approach for cardiac repair. OBJECTIVE:To evaluate the effect of polypyrrole-chitosan conductive composite hydrogel on cardiac function in rats with myocardial ischemia-reperfusion injury. METHODS:The polypyrrole-chitosan conductive composite hydrogel was prepared by chemical oxidative polymerization.The micromorphology,biocompatibility and conductivity of the hydrogels were characterized.Thirty adult SD rats were selected to establish a myocardial ischemia-reperfusion injury model by clamping the left anterior descending branch of the heart and then releasing it.After 21 days of modeling,the rats were divided into three groups by the random number table method:Normal saline was injected into the left ventricular infarction area and infarction margin area in the blank group.Chitosan hydrogel was injected into the left ventricular infarction area and infarction margin area in the ordinary hydrogel group.The polypyrrole-chitosan conductive composite hydrogel was injected into the left ventricular infarction area and infarction margin area,with 10 rats in each group.The corresponding time points after modeling were set,and cardiac mechanical function(echocardiogram,pressure-volume analysis),cardiac electrophysiology(electrocardiogram,programmed electrical stimulation,optical mapping technology,microelectrode array technology,eight-lead electrocardiogram,and electrical resistivity of the scar area)and cardiac histology were detected. RESULTS AND CONCLUSION:(1)There were a lot of pores on the surface of the conductive composite hydrogel,and the conductivity was(3.19±0.03)×10-3 mS/cm,which had good biocompatibility co-cultured with smooth muscle cells.(2)After 105 days of modeling,echocardiogram and pressure-volume analysis showed that compared with the blank group and the ordinary hydrogel group,the conductive composite hydrogel could significantly improve the contractile function of the heart of rats with myocardial ischemia-reperfusion injury.The results of electrocardiogram,programmed electrical stimulation,optical mapping technology,microelectrode array technology,eight-lead electrocardiogram,and electrical resistivity of the scar area examination at 105 days after modeling displayed that,compared with the blank group and the ordinary hydrogel group,the conductive composite hydrogel could significantly improve the electrical conduction function of the heart of rats with myocardial ischemia-reperfusion injury and reduce the occurrence of arrhythmia.Masson staining of heart tissue at 105 days after modeling exhibited that there were different degrees of fibrosis in the myocardial infarction area of the three groups.Compared with the normal saline group and the ordinary hydrogel group,the conductive hydrogel group had more normal myocardial tissue and less fibrosis in the myocardial infarction area.(3)The results verify that polypyrrole-chitosan conductive composite hydrogel may promote the repair of infarcted heart after ischemia-reperfusion injury by increasing the electrical conduction velocity of infarct scar area tissue,increasing scar thickness,enhancing synchronous cardiac contraction,and reducing damaged tissue.

Objective To establish and verify a universal and stable potency test method in vitro for SARS-CoV-2 mRNA vaccine,so as to use it for the quality control of SARS-CoV-2 mRNA vaccine.Methods ELISA kits that could bind well to S protein of SARS-CoV-2 variants,as well as transfected cells,cell plating concentrations and doses for transfection were screened,and then a potency test method for SARS-CoV-2 mRNA vaccine in vitro was established and verified.Results An ELISA kit was found with good binding ability to S protein of each variant,and HEK293T cells were determined as the transfection cells,with the plating concentration of 2.5 × 10~5 cells/mL and the transfection dose of 4 μg/well in the 6-well plate.An universal and stable potency test method for SARS-CoV-2 mRNA vaccine in vitro was established.The verification results showed that the method met the quality control needs.Conclusion The established potency test method in vitro for SARS-CoV-2 mRNA vaccine has good relative accuracy,linearity,intermediate precision and range,and can be applied to the quality control of SARS-CoV-2 mRNA vaccines

With the development of modern medical standards,autoimmune diseases and their associ-ated successive osteoporosis have received increasing attention in recent years.Patients with autoimmune diseases,due to the characteristics of the disease and the prolonged use of glucocorticoid hormone thera-py,may affect the bone formation and bone absorption of the patient,followed by severe successive osteo-porosis,thereby increasing the risk of osteoporotic vertebral fractures.Vertebral compression fractures of the spine are common fracture types in patients with osteoporotic fractures.Osteoporosis is a common complication after glucocorticoid therapy in patients with autoimmune diseases.Percutaneous vertebro-plasty(PVP)and percutaneous kyphoplasty(PKP)are minimally invasive operation and are commonly used surgical methods for the treatment of osteoporotic vertebral compression fractures.However,due to the operation of spinal puncture during the operation,there are serious surgical risks such as bone cement leakage,spinal epidural hemorrhage,subdural hemorrhage,and subarachnoid hemorrhage in both PVP and PKP.As a result,it is necessary to evaluate the patient's body before surgery carefully,especially in the case of blood coagulation.This article reports a case of autoimmune disease patient admitted to Peking University People's Hospital due to lumbar 4 vertebral compression fracture combined with Sj?gren's syn-drome.The patient's preoperative examination showed that the activated partial thromboplastin time(APTT)was significantly prolonged.After completing the APTT extended screening experiment and lu-pus anticoagulant factor testing,the multi-disciplinary team(MDT)of Peking University People's Hospi-tal jointly discussed the conclusion that the patient's test results were caused by an abnormal self-immuni-ty anti-copulant lupus(LAC).Based on the results of the laboratory examination,the patient was con-sidered to be diagnosed with combined antiphospholipid syndrome(APS).For such patients,compared with the patient's tendency to bleed,we should pay more attention to the risk of high blood clotting in the lower limbs of the patient,pulmonary clots and so on.With timely anti-coagulation treatment,the patient safely passed the peripheral period and was successfully discharged from the hospital.Therefore,for pa-tients with autoimmune diseases with prolonged APTT in the perioperative period,doctors need to careful-ly identify the actual cause and carry out targeted treatment in order to minimize the risk of surgical and perioperative complications and bring satisfactory treatment results to the patients.

Objective To construct a nonparametric proportional hazards (PH) model for mixed informative interval-censored failure time data for predicting the risks in heart transplantation surgeries. Methods Based on the complexity of mixed informative interval-censored failure time data, we considered the interdependent relationship between failure time process and observation time process, constructed a nonparametric proportional hazards (PH) model to describe the nonlinear relationship between the risk factors and heart transplant surgery risks and proposed a two-step sieve estimation maximum likelihood algorithm. An estimation equation was established to estimate frailty variables using the observation process model. I-spline and B-spline were used to approximate the unknown baseline hazard function and nonparametric function, respectively, to obtain the working likelihood function in the sieve space. The partial derivative of the model parameters was used to obtain the scoring equation. The maximum likelihood estimation of the parameters was obtained by solving the scoring equation, and a function curve of the impact of risk factors on the risk of heart transplantation surgery was drawn. Results Simulation experiment suggested that the estimated values obtained by the proposed method were consistent and asymptotically effective under various settings with good fitting effects. Analysis of heart transplant surgery data showed that the donor's age had a positive linear relationship with the surgical risk. The impact of the recipient's age at disease onset increased at first and then stabilized, but increased against at an older age. The donor-recipient age difference had a positive linear relationship with the surgical risk of heart transplantation. Conclusion The nonparametric PH model established in this study can be used for predicting the risks in heart transplantation surgery and exploring the functional relationship between the surgery risks and the risk factors.

Objective A competing endogenous RNA (ceRNA) regulatory network associated with long non-coding RNA (lncRNA) specific for lung adenocarcinoma (LUAD) was constructed based on bioinformatics methods, and the functional mechanism of actinfilament-associated protein 1-antisense RNA1 (AFAP1-AS1) in LUAD was analyzed, in order to provide a new direction for the study of LUAD therapeutic targets. Methods The gene chip of LUAD was downloaded from the Gene Expression Omnibus (GEO), and lncRNA and mRNA with differential expression between LUAD and normal tissues were screened using GEO2R online software, and their target genes were predicted by online databases to construct ceRNA networks and perform enrichment analysis. In cell experiments, AFAP1-AS1 was genetically knocked down and siRNA was constructed and transfected into LUAD cells A549 by cell transfection. CCK8, transwell, scratch assay and flow cytometry were used to detect the ability of cells to proliferate, invade, migrate and apoptosis. Results A total of 6 differentially expressed lncRNA and 494 differentially expressed mRNA were identified in the microarray of LUAD. The ceRNA network involved a total of 6 lncRNA, 22 miRNA, and 55 mRNA. Enrichment analysis revealed that mRNA was associated with cancer-related pathways. In cell assays, knockdown of AFAP1-AS1 inhibited cell proliferation, invasion, and migration, and AFAP1-AS1 promoted apoptosis. Conclusion In this study, we construct a lncRNA-mediated ceRNA network, which may help to further investigate the mechanism of action of LUAD. In addition, through cellular experiments, AFAP1-AS1 is found to have potential as a therapeutic target for LUAD.

Objective To construct a nonparametric proportional hazards (PH) model for mixed informative interval-censored failure time data for predicting the risks in heart transplantation surgeries. Methods Based on the complexity of mixed informative interval-censored failure time data, we considered the interdependent relationship between failure time process and observation time process, constructed a nonparametric proportional hazards (PH) model to describe the nonlinear relationship between the risk factors and heart transplant surgery risks and proposed a two-step sieve estimation maximum likelihood algorithm. An estimation equation was established to estimate frailty variables using the observation process model. I-spline and B-spline were used to approximate the unknown baseline hazard function and nonparametric function, respectively, to obtain the working likelihood function in the sieve space. The partial derivative of the model parameters was used to obtain the scoring equation. The maximum likelihood estimation of the parameters was obtained by solving the scoring equation, and a function curve of the impact of risk factors on the risk of heart transplantation surgery was drawn. Results Simulation experiment suggested that the estimated values obtained by the proposed method were consistent and asymptotically effective under various settings with good fitting effects. Analysis of heart transplant surgery data showed that the donor's age had a positive linear relationship with the surgical risk. The impact of the recipient's age at disease onset increased at first and then stabilized, but increased against at an older age. The donor-recipient age difference had a positive linear relationship with the surgical risk of heart transplantation. Conclusion The nonparametric PH model established in this study can be used for predicting the risks in heart transplantation surgery and exploring the functional relationship between the surgery risks and the risk factors.