1.Research on The Construction and Application of Multiple Fluorescence Amplification System for Three Kinds of Stains
Yi-Fan BAI ; He-Miao ZHAO ; Jing CHEN ; Hong-Di LIU ; Rui-Qin YANG ; Chong WANG
Progress in Biochemistry and Biophysics 2025;52(4):982-994
ObjectiveA multiplex amplification system was constructed based on the capillary electrophoresis platform for simultaneous detection of saliva, semen, and vaginal secretions using tissue-specific RNA markers. The aim of this study is to identify the tissue origin of suspicious body fluid stains found at crime scenes and determine whether the body fluid stains at the crime scene are one or several types among saliva, semen, and vaginal secretions. MethodsThirty saliva samples, forty semen samples, and forty vaginal secretion samples (half from 2015 and half from 2024) were collected from healthy adult volunteers. Through primer designing, system formulation, and PCR condition optimization, a multiplex fluorescent amplification system was constructed. The specificity, sensitivity, and detection ability for mixed samples of this system were investigated, and it was tested using real crime scene materials. In the primer design stage, to reduce the requirements for RNA template quality, the amplification products were set within 80-300 bp. In the system formulation stage, dominant and subordinate primers were mainly considered. By reducing the concentration of dominant primers and increasing that of subordinate primers, a capillary electrophoresis spectrum with an appropriate peak height ratio was finally obtained. Additionally, gradient experiments were designed to adjust the concentrations of PCR reagents and PCR amplification conditions, and multiple versions of DNA amplification enzymes were optimized to achieve the best experimental results. ResultsThrough statistical analysis, there was no significant difference in the capillary electrophoresis of the 3 types of body fluid samples from the two years (2015 and 2024), demonstrating that the sample preservation method in this study can preserve samples for a relatively long time. The composite amplification system constructed in this study exhibited high specificity for all 3 types of body fluid, with no cross-reactions between the markers of each type of body fluid. The minimum detection thresholds for the 3 types of body fluid reached 0.002 9, 0.001 5, and 0.42 mg/L, respectively. This system also had a high degree of discrimination for mixed samples, especially for semen-saliva mixtures, where each body fluid marker could still be successfully detected when the concentration ratio of semen to saliva was 100:1. Meanwhile, in the two actual cases presented in this article, the application of this composite amplification system performed outstandingly. ConclusionThe composite amplification detection system constructed in this study can achieve the correct screening of saliva, semen, and vaginal secretions, overcoming the problems such as low specificity and sensitivity of marker tests and unbalanced RFU values of each marker in previous studies. The specificity and sensitivity meet the practical work requirements, and the operation is simple. It provides an analytical and identification method for body fluid stains in actual case and is applicable to the identification of the tissue origin of biological evidence at crime scenes involving sexual assault, indecent assault, and other criminal acts. In the future, more types of body fluid markers will be screened to expand the types of body fluids detected by the system, and body fluid-specific cSNP and cInDel genetic markers will be introduced to infer the sources (individuals and types) of mixed and complex stains more accurately.
2.Mechanism of inhibitory effect of total flavonoids from Taraxacum mongolicum on obesity in mice by regulating intestinal flora
Yixue GAO ; Lin GUO ; Linyan LANG ; Jing WU ; Haoyang WANG ; Jing YANG ; Mingsan MIAO ; Zhanzhan LI
China Pharmacy 2025;36(3):293-299
OBJECTIVE To investigate the mechanism of the inhibitory effect of total flavonoids from Taraxacum mongolicum on high-fat diet-induced obesity in mice through modulation of intestinal flora. METHODS Twenty-four C57BL/6J mice were randomly divided into blank group, model group and T. mongolicum total flavonoid group, with 8 mice in each group. Except for the blank group, the other 2 groups were given a high-fat diet, while T. mongolicum total flavonoid group was given T. mongolicum total flavonoid [400 mg/(kg·d)] intragastrically, once a day, for 8 consecutive weeks. During the experiment, the food intake of each group of mice was recorded. After the last medication, the body mass, fat weight, blood lipid level and pathological changes of liver and epididymal fat in mice were evaluated to observe the effect of T. mongolicum total flavonoid on the treatment of obesity in mice. The changes in abundance and structure of intestinal flora in mice were detected by amplicon sequencing; the effects of T. mongolicum total flavonoids on fat metabolism related genes were analyzed by qPCR. RESULTS Compared with model group, the body weight of mice in T. mongolicum total flavonoids group was decreased significantly (P<0.05); the levels of total lipid cholesterol, triglycerides, and LDL cholesterol were all decreased significantly (P<0.01), and the level of HDL cholesterol was increased significantly (P<0.01); the fat indexes of inguinal white adipose tissue and epididymal white wind_lz@hactcm.edu.cn adipose tissue were significantly reduced (P<0.05); significant improvement in hepatocellular steatosis and adipose cytopathy were significantly improved; mRNA expressions of COX7A1 and COX8B were significantly upregulated (P<0.05). The results of bacterial colony detection showed that compared with the model group, there was a rising trend in the diversity of the bacterial colony in T. mongolicum total flavonoids group, and the Sobs index characterization and β diversity were increased significantly (P<0.05). Relative abundances of Blautia, norank_f_Ruminococcaceae, Bilophila, Alistipes, classified_f_Ruminococcaceae, Parabacteroides, norank_f_Desulfovibrionaceae, Anaerotruncus were significantly up-regulated(P<0.05), while those of Faecalibaculum, Erysipelatoclostridium, GCA-900066575, Tuzzerella, Lactobacillus, norank_f_norank_o_RF39, achnospiraceae_FCS020_group were significantly down-regulated (P<0.05). CONCLUSIONS T. mongolicum total flavonoids can reduce body mass, fat weight and blood lipid levels, and repair the pathological damage to liver and epididymal fat in obese mice, which is related to improving intestinal flora disorders caused by high-fat diet.
3.Two new glycosides from the Citri Sarcodactylis Fructus
Jing-jing MIAO ; Ge-ge XIA ; Ge-ge ZHAO ; Yu-zhong ZHENG ; Yan-zhi WANG
Acta Pharmaceutica Sinica 2025;60(1):196-200
Six compounds were isolated from the ethyl acetate fraction of
4.Pharmaceutical care of a patient with inflammatory bowel disease and atrial fibrillation
Jinfa CAO ; Ling ZHONG ; Miao HE ; Jing TIAN
Journal of Pharmaceutical Practice and Service 2025;43(10):525-528
Objective To investigate the disease characteristics, potential drug interactions, and key points of pharmaceutical care in patients with inflammatory bowel disease and atrial fibrillation, so as to provide a reference for rational clinical medication use. Methods Through involvement in the pharmaceutical care of a patient with inflammatory bowel disease and atrial fibrillation, and by reviewing relevant literature, the clinical pharmacist assessed the rationality of the medication regimen, proposed medication reconciliation recommendations, and assisted the multidisciplinary team in optimizing the treatment plan. Results Following the intervention, the patient's inflammatory bowel disease-related symptoms were improved significantly, atrial fibrillation was effectively controlled, and no significant adverse drug reactions were observed. Conclusion There exists a complex bidirectional interaction between inflammatory bowel disease and atrial fibrillation in terms of pathological mechanisms and pharmacotherapy. Through individualized medication reconciliation and multidisciplinary pharmaceutical care, safe and effective long-term management of those patients can be achieved.
5.Nutritional status and influencing factors in elderly patients with chronic renal insufficiency
Miao ZHU ; Manman LYU ; Haichuan YUAN ; Juantang ZHAO ; Xiujuan WU ; Jing TAO
Journal of Public Health and Preventive Medicine 2025;36(6):171-175
Objective To assess the nutritional status in elderly patients with chronic renal insufficiency (CRI) and reveal the key factors affecting the nutritional status. Methods A total of 310 elderly patients with CRI who received hospitalization treatment and outpatient follow-up in the hospital from January 2021 to June 2024 were selected as the investigation subjects. The nutritional status of patients was evaluated by mini-nutritional assessment (MNA) questionnaire, and the nutritional status and dietary structure of patients were comprehensively evaluated by anthropometric indicators [height, weight, body mass index (BMI), upper arm circumference, calf circumference], biochemical indicators [serum albumin (ALB), prealbumin (PA), hemoglobin (Hb), transferrin (TF)] and 24-hour dietary review method. According to the investigation results of nutritional status, the patients were divided into good nutrition group (MNA score≥24 points), nutritional risk group (MNA score of 17-23.5 points) and malnutrition group (MNA score<17 points). Univariate analysis was adopted to screen the potential influencing factors of elderly CRI. Multivariate logistic regression model was applied to analyze the influencing factors of malnutrition in elderly CRI patients. Results Among the 325 questionnaires were distributed, but only 310 valid questionnaires were recovered, with an effective recovery rate of 95.38%. Investigation results revealed that among the 310 patients, 29.35% (91 cases) had good nutritional status, and 42.26% (131 cases) had nutritional risk, and 28.39% (88 cases) had malnutrition. Univariate analysis indicated that there were statistical differences in BMI, CRI staging, serum ALB, PA, Hb, TF, protein intake and total calorie intake among the good nutrition group, the nutritional risk group and the malnutrition group (P<0.05). Multivariate logistic regression analysis suggested that low BMI (OR=0.903, 95%CI: 0.867-0.941), high CRI stage (OR=1.091, 95%CI: 1.053-1.130), low serum ALB (OR=0.907, 95%CI: 0.867-0.948), PA (OR=0.918, 95%CI: 0.888-0.949), Hb (OR=0.944, 95%CI: 0.909-0.997), TF (OR=0.912, 95%CI: 0.874-0.952), insufficient protein intake (OR=0.924, 95%CI: 0.882-0.969) and insufficient total calorie intake (OR=0.938, 95%CI: 0.909-0.968) were influencing factors for malnutrition in elderly patients with CRI (all P<0.05). Drawing ROC curve of malnutrition in elderly patients with CRI according to the prediction probability of logistic regression model found that the AUC, sensitivity, specificity, 95%CI and Youden index were 0.976, 93.18%, 92.34%, 0.953-0.990 (P<0.05) and 0.855. Conclusion The incidence rate of malnutrition is high in elderly patients with CRI, and is mainly affected by factors such as low BMI, high CRI stage, low serum ALB, PA, Hb and TF levels and insufficient protein and total calorie intakes. In addition, logistic regression model has high predictive value and can provide a reference for early clinical identification of high-risk population with malnutrition among elderly patients with CRI.
6.Optimization and practice of occupational education curriculum system for nuclear emergency medical rescue
Jiajin LIN ; Jing LI ; Wei HE ; Shenglong XU ; Dalu LIU ; Wei ZHANG ; Juan GUO ; Xia MIAO ; Yan ZHOU
Chinese Journal of Medical Education Research 2024;23(7):931-935
Nuclear emergency medical rescue is one of the important courses of military medical professional education, and improving the training level of nuclear emergency medical rescue plays a very important role in improving post competency of trainees. Based on the problem of disconnection between "teaching" and "need" in the occupational education of nuclear emergency medical rescue in the past, this study proposes the curriculum goal of "the combination of three abilities" and performs the optimization and practice of the occupational education curriculum system of nuclear emergency medical rescue from the aspects of curriculum setting, curriculum content, teaching methods, and assessment and evaluation. The results show that the new curriculum system can significantly improve the comprehensive ability of nuclear emergency medical rescue among trainees and better meet the requirements for their posts, thereby playing an important role in cultivating high-quality military medical talents in nuclear emergency medical rescue.
7.Exploring mechanism of action of Xihuang pills against hyperplasia of mammary glands based on quantitative proteomics
Rui TAO ; Jing-Rui WANG ; Jun-Liang WANG ; Xue-Li MA ; Juan-Xia SUN ; Guang-Miao GAO ; Qi-Rui FAN ; Tao HAN
Chinese Pharmacological Bulletin 2024;40(9):1641-1648
Aim To clarify the differential proteins of mammary tissues in Xihuang pills(XHP)against hy-perplasia of mammary glands(HMG)based on quanti-tative proteomics technology and validate them,and to explore the mechanism of action.Methods SD rats were randomly divided into blank group,model group and XHP group,with 10 rats in each group.Except for the blank group,estrogen and progesterone were injec-ted intramuscularly to establish a rat model of mamma-ry hyperplasia for 30 d.After XHP was administered for 14 d,the rats in each group were observed to have morphological changes in the apparent morphology of the mammary tissues,and pathological changes in the mammary tissues were stained with hematoxylin-eosin staining(HE),and the differentially expressed pro-teins(DEPs)in the groups were screened by quantita-tive proteomics technology and subjected to bioinforma-tics analysis,and Western blot to verify the key DEPs.Results Compared with the model group,the appar-ent pathological morphology of the XHP group was sig-nificantly improved,the diameter of the nipple height of the rats was significantly reduced(P<0.01),and the degree of histopathology was significantly allevia-ted.Quantitative proteomics identified 4,299 DEPs in mammary tissue,and bioinformatics analysis of 14 DEPs with consistent changes between the XHP group and the blank group relative to the model group re-vealed that they were related to the regulation of mus-cular systemic processes,regulation of muscle contrac-tion,DNA replication,and pre-initiation of DNA repli-cation.Western blot results showed that,compared with the model group,rat mammary tissue of the XHP group showed significantly lower levels of ACLY and ALDOC protein expression levels were significantly re-duced and BIN1 protein expression levels were signifi-cantly increased(P<0.01).Conclusions XHP may exert its anti-mammary hyperplasia effect through the regulation of BIN1,ACLY and ALDOC protein lev-els,the regulation of DNA replication,the regulation of pre-initiation of DNA replication and muscular sys-temic processes,and the regulation of muscle contrac-tion.
8.Ginkgo biloba extract activates Nrf2/ARE pathway to improve vascular endothelial dysfunction induced by chronic intermittent hypoxia in rats
Sheng-Yong SI ; Hong-Man LI ; Si-Si MIAO ; Xiao HAN ; Zhi-Jing LI ; Chao-Jun WEI ; Da-Nan LIU
Chinese Pharmacological Bulletin 2024;40(10):1837-1844
Aim To investigate the effects of Ginkgo biloba extract(GBE)on vascular endothelial dysfunc-tion induced by chronic intermittent hypoxia(CIH)in rats and its related mechanisms.Methods The CIH rat model was established,and 50 and 100 mg·kg-1 GBE was administered by intragastric administration.The systolic blood pressure(SBP)of the tail artery was detected in each group.HE staining was used to detect the morphology of aorta tissue.DAF-FM DA staining and nitric reductase assay were used to detect NO levels.ELISA was used to detect serum ET-1,TNF-α and IL-6 levels.DHE staining was used to de-tect reactive oxygen species(ROS)levels of aortic tis-sue.Kits were used to detect the serum levels of MDA,SOD and GSH-Px.Western blot was used to detect the levels of VCAM-1,ICAM-1,nucleus Nrf2,HO-1 and NQO1 of aortic tissue.Results GBE sig-nificantly decreased the levels of SBP,ET-1,ROS,MDA,VCAM-1,ICAM-1,TNF-α and IL-6,and sig-nificantly increased the levels of NO,SOD,GSH-Px,nuclear Nrf2,HO-1 and NQO1 in CIH rats.GBE sig-nificantly improved the histomorphology of aorta in CIH rats.Conclusions GBE can improve vascular endo-thelial dysfunction and reduce blood pressure in CIH model rats.The mechanism may be related to the acti-vation of Nrf2/ARE pathway and the inhibition of oxi-dative stress and inflammation by GBE.
9.Microgravity-mediated Notch1 signaling pathway affects bone homeostasis by regulating macrophage polarization
Jing XU ; Jian GUO ; Yonggui LUO ; Daxing LI ; Ying TANG ; Baojia LOU ; Miao PENG ; Yong ZHENG
Chinese Journal of Immunology 2024;40(8):1625-1633
Objective:To investigate the effect of microgravity-mediated Notch1 signaling on macrophage polarization on bone homeostasis.Methods:The animal model was constructed by tail-limb suspension(HLS)to simulate the microgravity environment.The animals were grouped into Control group,HLS group,HLS+NC group,HLS+si group,HLS+rhNF-κB group.ELISA was used to detect the content of TNF-α and IL-1β in serum of rats.TUNEL staining was used to detect the apoptosis of bone tissue.Immunofluo-rescence was used to detect the polarization of macrophages in bone tissue.The rat osteoblast CP-R091 microgravity model was con-structed by simulating the microgravity environment with a rotating wall bioreactor.The cell experiments were divided into Control group,HLS group,HLS+NC group,HLS+si group,HLS+rhNF-κB group.CCK-8 test was used to detect the proliferation activity of cells in each group,and AO test was used to test the apoptosis rate of cells in each group.PCR was used to detect the expression of os-teogenesis-related genes in bone tissues and cells.Western blot was used to detect the expression of Notch1,hair division-related en-hancer-1(HES-1),and Notch pathway ligand 1(Jagged1)in bone tissues and cells of each group.Results:Compared with control group,the contents of TNF-α and IL-1β in the serum of the rats in the HLS group,the apoptosis rate,and the proportion of M1 macro-phages were significantly increased.Compared with HLS group,the HLS+si group could obviously partially reverse the change trend of the above parameters,while HLS+rhNF-κB group significantly changed the above parameter values.Compared with control group,the proliferation activity of the cells in the HLS group was significantly reduced,and the apoptosis rate was significantly increased.Com-pared with HLS group,the HLS+si group could obviously partially reverse the change trend of the above parameters,while the HLS+rhNF-κB group made the above parameter values worse.The expressions of the osteogenesis-related genes collagen type Ⅰ(COL1),osteocalcin(OCN)and Runt-related transcription factor 2(RUNX2)in bone tissues and cells in the microgravity environment were significantly decreased,while the expressions of Notch-1,Hes-1 and Jagged1 were significantly increased,and the differences were statistically significant(all P<0.05).Conclusion:Microgravity-mediated Notch1 signaling regulates M1/M2 polarization of macro-phages,participates in cell proliferation and apoptosis in bone tissue,and affects the progress of bone homeostasis.
10.A re-evaluation of the effectiveness of exercise interventions in patients with amyotrophic lateral sclerosis
Jing SHAN ; Yajie MIAO ; Jia LIU ; Jinju SUN ; Xue MENG ; Mengxue YANG
Chinese Journal of Modern Nursing 2024;30(28):3852-3858
Objective:To re-evaluate the effectiveness of exercise interventions in patients with amyotrophic lateral sclerosis (ALS) by conducting a systematic review and provide insights for the implementation and continuous improvement of exercise intervention strategies.Methods:A comprehensive search was conducted across databases, including CNKI, Wanfang, VIP, SinoMed, PubMed, Embase, Cochrane Library, Web of Science, CINAHL, Campbell Collaboration, and the Joanna Briggs Institute Evidence-Based Health Care Center Database. The search covered literature from the inception of the databases until December 19, 2023. Two researchers trained in evidence-based nursing independently screened the literature. The methodological quality of the included studies was assessed using the AMSTAR 2 tool, and the quality of evidence was graded using the GRADE system.Results:Eight systematic reviews were included. The AMSTAR 2 evaluation revealed one review of moderate quality, six of low quality, and one of very low quality. GRADE assessment of 33 outcome indicators showed that one piece of evidence was of moderate quality, 24 were of low quality, and eight were of very low quality.Conclusions:Exercise interventions may improve overall functional outcomes in ALS patients without exacerbating fatigue. However, due to the limitations in the quantity and quality of the included studies, further large-scale, high-quality randomized controlled trials are needed to verify the effects of exercise interventions in ALS patients.


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