Objective:To establish a genomic nucleic acid mass spectrometry detection platform for allelic risk associated with Alzheimer's disease.Methods:Whole blood samples of 61 patients diagnosed as Alzheimer's disease in the Affiliated Brain Hospital of Guangzhou Medical University from December 28th, 2023 to 31st, March 2024 were collected and deoxynucleic acid (DNA) was extracted, including 22 males and 39 females, aged (67.36 ± 8.18) years old. After screening out 17 risk gene loci in Chinese population, multiplex polymerase chain reaction primers, single-base extension primers and Sanger sequencing primers were designed. Ten samples were used for primer optimization and debugging through Sanger sequencing and time-of-flight mass spectrometry to establish a detection system. The remaining samples were genotyped using a time-of-flight mass spectrometer and verified by Sanger sequencing for accuracy evaluation. Five samples were selected for gradient dilution and then subjected to time-of-flight mass spectrometry detection to evaluate the detection limit. Three clinical samples, one case of Escherichia coli and one case of Staphylococcus aureus genomic DNA samples were selected for cross-reaction research. The anti-interference ability of the detection system was evaluated against hemolysis, chylous substances and conventional anticoagulants in the samples. Two samples, one wild and one homozygous mutation sample with representative peak shapes, were selected to evaluate the anti-interference ability. Four samples containing the common genotypes of all gene loci in the system were selected and repeated 10 times to evaluate the precision.Results:The minimum intensity of single-base extension primers on mass spectrometry is greater than half of the maximum intensity. All 17 risk gene loci screened were successfully typed. The time-of-flight mass spectrometry detection results of 1,037 loci from 61 samples showed that the genotyping detection rate was 100%. The genotypes of the 20 DNA samples were completely consistent with the results of Sanger sequencing, with an accuracy rate of 100%. The mass spectrometry detection results of five samples after gradient dilution indicated that the low detection limit was 5 ng of DNA. The reaction system has a strong anti-interference ability against hemolysis of samples, chylous substances, conventional anticoagulants and DNA cross-contamination. Homologous allele interference and no cross-reaction between the bacterial genome and 17 gene loci do not affect the risk genome detection results. The results of 10 repeated mass spectrometry tests on 4 samples showed that the precision was 100%.Conclusion:The genomic detection system of Alzheimer's disease risk has been successfully established to provide an auxiliary mean for disease diagnosis and risk assessment.

Objective:To establish a genomic nucleic acid mass spectrometry detection platform for allelic risk associated with Alzheimer's disease.Methods:Whole blood samples of 61 patients diagnosed as Alzheimer's disease in the Affiliated Brain Hospital of Guangzhou Medical University from December 28th, 2023 to 31st, March 2024 were collected and deoxynucleic acid (DNA) was extracted, including 22 males and 39 females, aged (67.36 ± 8.18) years old. After screening out 17 risk gene loci in Chinese population, multiplex polymerase chain reaction primers, single-base extension primers and Sanger sequencing primers were designed. Ten samples were used for primer optimization and debugging through Sanger sequencing and time-of-flight mass spectrometry to establish a detection system. The remaining samples were genotyped using a time-of-flight mass spectrometer and verified by Sanger sequencing for accuracy evaluation. Five samples were selected for gradient dilution and then subjected to time-of-flight mass spectrometry detection to evaluate the detection limit. Three clinical samples, one case of Escherichia coli and one case of Staphylococcus aureus genomic DNA samples were selected for cross-reaction research. The anti-interference ability of the detection system was evaluated against hemolysis, chylous substances and conventional anticoagulants in the samples. Two samples, one wild and one homozygous mutation sample with representative peak shapes, were selected to evaluate the anti-interference ability. Four samples containing the common genotypes of all gene loci in the system were selected and repeated 10 times to evaluate the precision.Results:The minimum intensity of single-base extension primers on mass spectrometry is greater than half of the maximum intensity. All 17 risk gene loci screened were successfully typed. The time-of-flight mass spectrometry detection results of 1,037 loci from 61 samples showed that the genotyping detection rate was 100%. The genotypes of the 20 DNA samples were completely consistent with the results of Sanger sequencing, with an accuracy rate of 100%. The mass spectrometry detection results of five samples after gradient dilution indicated that the low detection limit was 5 ng of DNA. The reaction system has a strong anti-interference ability against hemolysis of samples, chylous substances, conventional anticoagulants and DNA cross-contamination. Homologous allele interference and no cross-reaction between the bacterial genome and 17 gene loci do not affect the risk genome detection results. The results of 10 repeated mass spectrometry tests on 4 samples showed that the precision was 100%.Conclusion:The genomic detection system of Alzheimer's disease risk has been successfully established to provide an auxiliary mean for disease diagnosis and risk assessment.

OBJECTIVE To explore the mechanism of joint injection of Caulophyllum robustum Maxim in the treatment of rheumatoid arthritis(RA).METHODS The targets of main saponins in C.robustum Maxim were obtained from Swiss Target Prediction,and the RA treatment targets collected from the GeneCards and OMIM database were intercrossed to establish an interaction network based on network pharmacology.Gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed.RA model was established by injecting complete Freund's adjuvant into the back of rabbits for verification.The arthritis index score,knee diameter and pain threshold of rabbits were compared.Pathological examination of rabbit synovial tissue was carried out.The levels of tumor necrosis factor α(TNF-α),interleukin-1β(IL-1β)and IL-6 in rabbit serum and synovial fluid were detected.The phosphorylation levels of tyrosine protein Janus kinase 2(JAK2)and signal transducer and activator of transcription 3(STAT3)proteins in rabbit synovium were detected.RESULTS Network pharmacology identified 143 intersection targets between the drug and RA.After the construction of the"drug-component-target"network,the core components of the network were echinocystic acid,oleanolic acid,hederagenin,cauloside A and cauloside C,etc.Additionally,the top 10 core targets of PPI network were SRC,STAT3,MAPK1,EGFR,PIK3CA,MAPK3,GRB2,JUN,PTPN11 and JAK2.The results of KEGG pathway analysis showed that the JAK/STAT signaling pathway was mainly involved in the treatment of RA by joint injection of C.robustum Maxim.Results of validation test showed that compared with model group,joint injection of C.robustum Maxim could reduce the swelling of rabbit knee joint,relieve the hyperplasia of synovial layer,reduce the hyperplasia of lower connective tissue,and reduce the number of inflammatory cells and capillaries.The arthritis index score(excluding low-dose group of C.robustum Maxim),knee diameter,the levels of TNF-α,IL-1β and IL-6 in serum and synovial fluid,and the protein phosphorylation levels of JAK2 and STAT3 were decreased significantly(P＜0.05 of P＜0.01),while the pain threshold were reduced significantly(P＜0.01).CONCLUSIONS The core components that may alleviate the inflammatory response of RA in joint injection of C.robustum Maxim could include echinocystic acid,oleanolic acid,hederagenin,cauloside A,and cauloside C.Its mechanism may be related to the inhibition of JAK/STAT signaling pathway and the reduction of inflammatory responses.

Objective To study the effect and mechanism of the Fosinopril combined with Fenofibrate on the prevent?ing of diabetic retinopathy. Methods A total of 150 viripotent ICR mice(100 male mice, 50 female mice) were randomly di?vided into five groups(n=30), including A group (Sham group), B group (Model group), C group [Fosinopril prevented group, 20 mg/(kg·d)], D group [Fenofibrate prevented group, 400 mg/(kg·d)] and E group (Fosinopril combined with Fenofibrate pre?vented group). The expression levels of Bax and Bcl-2 gene mRNA were determined by RT-PCR method. TUNEL staining method was used to detect the apoptosisi of retinal cells. Results The Bcl-2 mRNA of A group, Bax mRNA of B group were higher than those of other four groups. Bcl-2 mRNA of E group was higher than that of C group and D group, while the Bax mRNA was lower than those of two groups(all P<0.05). The TUNEL index of B group was the highest than other groups, which of E group

OBJECTIVE:To explore the regularity of relative correction factor (RCF) in multi-components by single marker (QAMS). METHODS:With the source of CNKI,literature retrieval was used to collect the relevant documents of multi-compo-nents by single marker (QAMS) and extract the calculated values by quantitative analysis of multi-components by single marker QAMS and measured values by external standard method (ESM),SPSS 19.0 software was used to calculate the percentage value (RSD,%) of relative standard deviation between calculated values and measured values,and the correlation between RSD and RCF was analyzed by drawing a scatter plot. RESULTS:Most of RCF values distributed in the range of >0.62-1.53,accounting for 76.7%to all data. Calculated values and measured values showed high similarity in this range and the RSD<3.0%data account-ed for 88.8% to all data,which had good credibility. Meanwhile,the similarity of calculated values and measured values was influ-enced by the differences of structure and physicochemical property of parent nucleus between other components under measured and internal references. The high degree of similarity between calculated values and measured values were decisive by very similar struc-tures of parent nucleus;the low degree was decisive by discrepant structures;and the low degree was also decisive by very similar structures and discrepant physicochemical property of parent nucleus. CONCLUSIONS:RCF will affect the credibility of content de-termination results for components under measured in QAMS,it is necessary to establish database of RCF,RSD and related param-eters.