1.Preparation of Pancreatic Polypeptide Nanobody and the Analysis of Binding Activity
Hong-Rui REN ; Qiong JIA ; Jia-Qin WANG ; Jing-Jing TIAN ; Rong-Jie LI ; Hua-Hua HAO ; Jian-Li LI ; Zhi-Can QU ; Rui-Wen FAN
Chinese Journal of Biochemistry and Molecular Biology 2024;40(9):1273-1281
Pancreatic polypeptide(PP),a pancreatic hormone containing 36 amino acids,plays impor-tant roles in the diagnosis and evaluation of pancreatic function,injury and diseases.In this study,a phage nanobody library against PP was constructed to screen specific PP nanobodies,which would be used to evaluate whether they have binding activity with PP antigen.After PP antigen with high purity was prepared by prokaryotic expression system,it was used to immunize alpaca to construct the nanobody li-brary against PP with high storage capacity and high abundance,from which 8 strains of PP nanobodies were obtained by phage display.One of nanobody strain(PP-VHH)was selected to be expressed in a prokaryotic expression system,which was induced overnight by IPTG.After purification and identifica-tion,the antigen-antibody binding activity and PP level in serum were detected by indirect ELISA and Sandwich ELISA methods,respectively.The results showed that PP-VHH had binding activity with PP,which could be used to detect PP in chicken and human serum.The Sandwich ELISA methods with R2 of the fitting curve 0.9868 could be used to detect PP concentrations of 48-55 pg/mL in the serum of chick-ens,while the concentrations of PP in human serum varied significantly.In summary,PP-VHH screened from nanobody library against PP could detect PP in serum,which would supply the basis for evaluation of abnormal pancreatic function and diagnosis of relative disease.
2.Visualization of nasal powder distribution using biomimetic human nasal cavity model.
Jiawen SU ; Yan LIU ; Hongyu SUN ; Abid NAEEM ; Huipeng XU ; Yue QU ; Caifen WANG ; Zeru LI ; Jianhua LU ; Lulu WANG ; Xiaofeng WANG ; Jie WU ; Lixin SUN ; Jiwen ZHANG ; Zhigang WANG ; Rui YANG ; Li WU
Acta Pharmaceutica Sinica B 2024;14(1):392-404
Nasal drug delivery efficiency is highly dependent on the position in which the drug is deposited in the nasal cavity. However, no reliable method is currently available to assess its impact on delivery performance. In this study, a biomimetic nasal model based on three-dimensional (3D) reconstruction and three-dimensional printing (3DP) technology was developed for visualizing the deposition of drug powders in the nasal cavity. The results showed significant differences in cavity area and volume and powder distribution in the anterior part of the biomimetic nasal model of Chinese males and females. The nasal cavity model was modified with dimethicone and validated to be suitable for the deposition test. The experimental device produced the most satisfactory results with five spray times. Furthermore, particle sizes and spray angles were found to significantly affect the experimental device's performance and alter drug distribution, respectively. Additionally, mometasone furoate (MF) nasal spray (NS) distribution patterns were investigated in a goat nasal cavity model and three male goat noses, confirming the in vitro and in vivo correlation. In conclusion, the developed human nasal structure biomimetic device has the potential to be a valuable tool for assessing nasal drug delivery system deposition and distribution.
3.IL2rg-/- rats support prolonged infection of human RSV
Rui XIONG ; Yong WU ; Yanwei YANG ; Zhe QU ; Susu LIU ; Yuya WANG ; Liying MA ; Rui FU ; Yihong PENG ; Chunnan LIANG ; Changfa FAN
Acta Laboratorium Animalis Scientia Sinica 2024;32(1):17-24
Objective To overcome the limitations of existing human respiratory syncytial virus(hRSV)animal models,such as semi-permissiveness and short duration of infection,this study established an IL2rg gene knockout(IL2rg-/-)rat model using TALEN gene editing technology.Methods The animal model was infected with hRSV intranasally.Clinical characteristics,body weight,and temperature changes were observed over the infection period(0~35 days).The total viral loads in respiratory organs,such as the nasal tissue,trachea,and lungs,were measured at various time points(4,11,20,and 35 days post-infection).Pathological analysis was conducted on target organs at the endpoint of observation(35 days post-infection).Changes in peripheral blood T,B,NK,and NKT cells and various cytokines were assessed at various time points(4,20,and 35 days post-infection).Results(1)IL2rg/-knockout rats sustained high viral loads in the nasal cavity upon intranasal inoculation with hRSV.The average peak titer rapidly reached 1 × 1010 copies/g in nasal tissue and 1 × 107 copies/g up to 5 weeks post-infection.(2)However,no significant pathological changes were noted in nasal,tracheal,or lung tissues.(3)An increase was observed in the content of peripheral blood B cells in hRSV-infected IL2rg--rats.(4)IL-6 and MCP-1 were increased in the early stage of infection and then decreased at the end of the observation period.Conclusions This study established a new IL2rg-/-rat model using TALEN technology and found that this model effectively supported high-level replication and long-term infection of hRSV,providing a good basis for antiviral drug screening and in vivo efficacy evaluation of anti-hRSV antibodies.
4.The taste correction process of ibuprofen oral solution based on the combination of electronic tongue technology and artificial taste comprehensive evaluation
Rui YUAN ; Yun-ping QU ; Yan WANG ; Ya-xuan ZHANG ; Wan-ling ZHONG ; Xiao-yu FAN ; Hui-juan SHEN ; Yun-nan MA ; Jin-hong YE ; Jie BAI ; Shou-ying DU
Acta Pharmaceutica Sinica 2024;59(8):2404-2411
This experiment aims to study the taste-masking effects of different kinds of corrigent used individually and in combination on ibuprofen oral solution, in order to optimize the taste-masking formulation. Firstly, a wide range of corrigent and the mass fractions were extensively screened using electronic tongue technology. Subsequently, a combination of sensory evaluation, analytic hierarchy process (AHP)-fuzzy mathematics evaluation, and Box-Behnken experimental design were employed to comprehensively assess the taste-masking effects of different combinations of corrigent on ibuprofen oral solution, optimize the taste-masking formulation, and validate the results. The study received ethical approval from the Review Committee of the Beijing University of Chinese Medicine (ethical code: 2024BZYLL0102). The results showed that corrigent fractions and types were screened separately through single-factor experiments. Subsequently, a Box-Behnken response surface design combined with AHP and fuzzy mathematics evaluation was used to fit a functional model:
5.Changes in serum immunoglobulin levels in children with thalassemia who undergo repeated blood transfusions and their correlation with delayed hemolytic transfusion reactions
Xiaohong JIN ; Meikun HU ; Rui CHEN ; Lilan GAO ; Shuxia WANG ; Mengxing LYU ; Kexuan QU
Chinese Journal of Blood Transfusion 2024;37(5):548-555
Objective To study the changes in serum immunoglobulin levels in children with thalassemia who undergo repeated blood transfusions and explore their correlation with delayed hemolytic transfusion reactions(DHTR).Methods Serum samples from children with thalassemia who received blood transfusion treatment from June 2022 to April 2023(ob-servation group)and healthy children who underwent physical examination(control group)in our hospital were collected.The levels of serum immunoglobulins(IgG subtype,IgM,IgA,IgE and IgD)were detected using flow cytometry CBA multi-factor quantitative detection technology,and the differences between the two groups were compared.The children were divided into 4 groups according to different transfusion numbers:≤10 numbers,11-30 numbers,31-50 numbers and>50 numbers,and the differences between different blood transfusion numbers and serum immunoglobulin levels in each group were compared using one-way analysis of variance(ANOVA).Children with thalassemia with DHTR were in the hemolysis group,and children with thalassemia who did not experience DHTR were in the non-hemolysis group.The changes in serum immunoglobulins(IgG subtypes,IgM,IgA,IgE and IgD)between the two groups were compared to explore the correlation between serum immunoglobulins in thalassemia children with repeated transfusion and DHTR.Results The levels of IgG1,IgG3,IgG4 and IgA in the observation group were significantly higher than those in the control group,with the increase of(2.07±2.12),(0.67±2.03),(0.30±0.37)and(6.04±11.40)mg/mL,respectively,while the level of IgD in observation group was significantly lower than that in the control group,with a decrease of(0.03±0.01)mg/mL,P<0.05.No significant difference was noticed in IgG2,IgM and IgE between the groups(P>0.05).IgG1 and IgG4 both significantly increased with the number of blood transfusions.The IgG1 in the 4 groups increased sequentially as(0.30±0.62),(0.41±0.51)and(3.60±3.48)mg/mL,and IgG4 increased sequentially as(0.12±0.13),(0.22±0.07)and(0.21±0.38)mg/mL.IgG2,IgM and IgD showed a significant decrease,with IgG 2,IgM,and IgD in four groups decreased as(0.91±1.50),(0.14±0.10)and(0.05±0.05)mg/mL,respectively,showing significant differences with the number of blood transfusions(P<0.05).No sig-nificant difference was found in IgG3,IgA and IgE with different number of transfusions(P>0.05).IgG1,IgG3 and IgG4 in the hemolysis group were significantly higher than those in the non-hemolysis group,with an increase of(4.44±3.41),(0.73±1.26)and(0.52±0.40),respectively(P<0.05).IgD in the hemolysis group was significantly lower than that in the non-hemolysis group,with a decrease of(0.00±0.06)mg/mL,P<0.05.No significance was noticed in IgG2,IgM,IgA and IgE between the hemolysis group and the non-hemolysis group(P>0.05).Conclusion The serum immunoglobulin levels of children with thalassemia who undergo repeated blood transfusions are abnormal.There are differences in correlation between the number of blood transfusions and serum immunoglobulin levels among children with thalassemia who undergo repeated blood transfusions.The relevant serum immunoglobulins for DHTR in children with thalassemia who undergo repeated blood transfusions are IgG1,IgG3 and IgG4.
6.Ameliorative effect of ginsenoside Rh1 on kidney injury in diabetic mice through activation of Nrf2/HO-1 signaling pathway
Meng QU ; Rui HUANG ; Xinda JU ; Yuxin LIU ; Jichen XIA ; Jiaxin HUANG ; Chunyan YU ; Zhiheng DONG
Journal of Jilin University(Medicine Edition) 2024;50(6):1565-1571
Objective:To discuss the protective effect of ginsenoside Rh1(G-Rh1)on kidney injury in the diabetic mellitus(DM)mice,and to clarify its mechanism.Methods:The diabetic kidney disease(DKD)model was prepared by using the high-fat,high-sugar diet combined with intraperitoneal injection of streptozotocin(STZ).A total of 48 C57/BL6 model mice were randomly divided into model group,nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor ML385 group(ML385 group)(30 mg·kg-1),G-Rh1 group(30 mg·kg-1),and G-Rh1+ML385 group(30 mg·kg-1 G-Rh1+30 mg·kg-1 ML385),and there were 12 mice in each group.Additionally,12 C57/BL6 mice were selected as control group.After treated for 8 weeks,automatic analyzer was used to detect the levels of fasting blood glucose(FBG),blood urea nitrogen(BUN),and serum creatinine(Scr)in serum of the mice in various groups,as well as 24 h urinary protein(24 h UP)levels in urine,and the kidney index was calculated;kits were used to detect the activities of superoxide dismutase(SOD)and lactate dehydrogenase(LDH),and the levels of malondialdehyde(MDA)in kidney tissue of the mice in various groups;Western blotting method was used to detect the expression levels of Nrf2 and heme oxygenase-1(HO-1)proteins in kidney tissue of the mice in various groups.Results:Compared with control group,the levels of FBG and kidney indexes in serum of the mice in model group,ML385 group,and G-Rh1+ML385 group were significantly increased(P<0.01),and the level of FBG in serum of the mice in G-Rh1 group was significantly increased(P<0.01);compared with model group,the kidney index of the mice in ML385 group was significantly increased(P<0.05),while the levels of FBG and kidney index of the mice in G-Rh1 group were significantly decreased(P<0.05 or P<0.01);compared with G-Rh1 group,the level of FBG and kidney index of the mice in G-Rh1+ML385 group were significantly increased(P<0.01).Compared with control group,the levels of BUN and Scr in serum,and 24 h UP in urine of the mice in model group,ML385 group,G-Rh1 group,and G-Rh1+ML385 group were significantly increased(P<0.01);compared with model group,the level of BUN in serum and 24 h UP in urine of the mice in ML385 group were significantly increased(P<0.05),while the levels of BUN and Scr in serum,and 24 h UP in urine of the mice in G-Rh1 group were significantly decreased(P<0.01);compared with G-Rh1 group,the levels of BUN and Scr in serum,and 24 h UP in urine of the mice in G-Rh1+ML385 group were significantly increased(P<0.01).Compared with control group,the activities of SOD in kidney tissue of the mice in model group,ML385 group,G-Rh1 group,and G-Rh1+ML385 group were significantly decreased(P<0.01),while the levels of MDA and LDH activities were significantly increased(P<0.01);compared with model group,the activity of SOD in kidney tissue of the mice in ML385 group was significantly decreased(P<0.05),and the level of MDA was significantly increased(P<0.05);the activity of SOD in kidney tissue of the mice in of G-Rh1 group was significantly increased(P<0.01),and the level of MDA and activity of LDH were significantly decreased(P<0.01);compared with G-Rh1 group,the activity of SOD in kidney tissue of the mice in G-Rh1+ML385 group was significantly decreased(P<0.01),and the level of MDA and activity of LDH were significantly increased(P<0.01).Compared with control group,the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in model group,ML385 group,G-Rh1 group,and G-Rh1+ML385 group were significantly decreased(P<0.05 or P<0.01);compared with model group,the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in ML385 group and G-Rh1+ML385 group were significantly decreased(P<0.05),while the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in G-Rh1 group were significantly increased(P<0.01);compared with G-Rh1 group,the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in G-Rh1+ML385 group were significantly decreased(P<0.01).Conclusion:Ginsenoside Rh1 reduces the oxidative stress and improves the kidney function,providing protective effects on kidney injury in the DM mice,and its mechanism may be related to the activation of the Nrf2/HO-1 signaling pathway.
7.Study on the current situation and influencing factors of job involvement for employed nurses in military hospital.
Zhi Yan SUN ; Jing Rui QU ; Wan Hong WEI ; Lu Wen ZHANG ; Ya Jun YI ; Lu LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(3):204-209
Objective: To investigate the current situation of job involvement of nurses in military hospitals in Henan Province and analyze the influencing factors, so as to provide reference for improving the level of job involvement of military nurses. Methods: In February 2022, the employed nurses of 4 military hospitals in Henan Province were investigated by convenient sampling method. A total of 663 questionnaires were collected, including 632 valid questionnaires, with an effective recovery rate of 95.32%. The self-designed questionnaire was used to investigate the basic information of nurses, the Job Involvement Scale was used to investigate the job involvement of nurses, the Emotional Labor Scale for Nurses was used to investigate nurses' emotions, and the Work-Family Conflict Scale was used to investigate the work-family conflict of nurses. Independent sample t-test and univariate analysis of variance were used to compare the job involvement of military employed nurses with different demographic characteristics, Pearson correlation analysis was used to explore the correlation between emotional labor, work-family conflict and job involvement, and hierarchical regression analysis was used to explore the impact of relevant variables on the job involvement of military employed nurses. Results: The total average score of job involvement of military employed nurses was (3.68±1.13), and the scores of vitality, dedication and focus were (3.64±1.15), (3.74±1.25) and (3.67±1.21) respectively. The total score of emotional labor of nurses was 33-80 (62.95±8.12), with an average score of (3.93±0.51). The total score of work-family conflict was 18-94 (55.16±13.53), with an average score of (3.06±0.75). Professional emotional regulation, patient-centered emotional inhibition and standardized emotional play were positively related to the job involvement (r=0.46, 0.41, 0.22, P<0.01). Time-based conflict, stress-based conflict and behavior-based conflict had negative correlation with the job involvement (r=-0.12, -0.23, -0.20, P<0.01). In hierarchical regression analysis, after controlling demographic variables, emotional labor and work-family conflict accounted for 17.2% and 4.2% of the variation of job involvement. Conclusion: The job involvement of military employed nurses tends to be at a moderate level. Emotional labor and work-family conflict can significantly affect their job involvement.
United States
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Humans
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Hospitals, Military
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Family Conflict
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Surveys and Questionnaires
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Regression Analysis
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Nurses
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Job Satisfaction
8.Research progress in pH-responsive antibacterial nanomaterials for inhibiting oral plaque biofilms.
Shan Lin QU ; Shuo YU ; Xiao Lin MA ; Rui WANG
Chinese Journal of Stomatology 2023;58(3):292-297
pH-responsive antibacterial nanomaterials are a new type of nanomaterials that can selectively undergo structural changes and trigger drug release according to significant pH differences produced by the body under physiological and pathological conditions. The formation of acidic microenvironment in plaque is the key to the pathogenicity of oral plaque biofilm, which also creates conditions for the oral application of pH-responsive antibacterial nanomaterials. pH-responsive antibacterial nanomaterials can respond to changes in pH of plaque microenvironment and accurately control the release of antibacterial drugs, providing a new direction for improving drug efficacy and targeted antibacterial. The present article reviews the classification, mechanism and application of pH-responsive antibacterial nanomaterials in oral plaque inhibition.
9.Research progress in application of deep learning technology for fracture diagnosis
Chinese Journal of Trauma 2023;39(4):378-384
Fractures are mostly caused by direct or indirect external forces and may lead to death in worsened situations. Timely and accurate diagnosis of fractures is critical in determining the severity of injury. Medical imaging is an important diagnosis and evaluation tool for fractures, but uncertain imaging, excessive images and doctors′ subjective reading will weaken diagnosis efficiency and affect subsequent treatment. Whereas, fracture diagnosis models based on deep learning technology can effectively reduce missed diagnosis and misdiagnosis caused by unclear fracture imaging, showing good diagnostic potential. The authors reviewed the deep learning technology and its application in fracture diagnosis, hoping to offer new ideas for the research and application of deep learning technology in fracture diagnosis.
10.Mechanism of Yuejuwan in Prevention and Treatment of Psychological and Heart Diseases Based on Liver TMT Labeled Quantitative Proteomics
Hanwen ZHANG ; Jiaxiang YU ; Yan SHI ; Wenshun ZHANG ; Xueying HAN ; Huan ZHANG ; Chao QU ; Xinhui SHEN ; Xiande MA ; Rui YU ; You YU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(1):26-36
ObjectiveTo observe the effects of Yuejuwan in the treatment of psychological and heart diseases (PHD) and explore its mechanism. MethodThirty 6-week-old healthy male SPF AopE-/- mice and 10 homologous C57BL/6J mice were selected for the experiment. The 30 AopE-/- mice were divided into a model group, low-dose (7.58 g·kg-1·d-1) and high-dose (30.32 g·kg-1·d-1) Yuejuwan groups, with 10 mice in each group, and 10 C57BL/6J mice were assigned to the blank control group. Intragastrical administration lasted 12 weeks. During feeding, the PHD model was induced by chronic unpredictable mild stress (CUMS) combined with high-fat diet in mice. After intragastric administration, the behavioral results [open field test (OFT) and sucrose preference test (SPT)] of mice in each group, the content of aspartic transaminase (AST), alanine aminotransferase (ALT), 5-hydroxytryptamine (5-HT), noradrenaline (NE), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), and triglyceride (TG) in serum of mice detected by the automatic biochemical analyzer, the oil red O staining and HE staining of aorta and liver and Masson staining of myocardial tissues were used for model evaluation. Finally, liver TMT-labeled quantitative proteomics was used to explore the mechanism of action. ResultThe model mice showed obvious manifestations of depression, anxiety, loss of interest, and despair, manifest lipid deposition in the aorta and liver by pathological observation, and increased myocardial fibrosis in myocardial tissues. After intragastric administration of Yuejuwan, the above symptoms and indexes of the PHD model mice were improved. Compared with the blank control group, the model group showed decreased standing times, cumulative time in the central area, total moving distance, moving speed, and sucrose preference at week 12 (P<0.01). Compared with the model group, the Yuejuwan groups showed decreased indexes mentioned above (P<0.01). After sample collection, AST, ALT, and TG levels in the model group were higher (P<0.01) and the levels of 5-HT, NE, and HDL-C were lower than those in the blank control group (P<0.01). The results of liver TMT labeled quantitative proteomics suggested that the PHD model mainly caused the changes in protein expression levels such as ApoE, UGT1A5, and FASN in mice,involving acetyl CoA metabolism,response to bacteria,cellular amino acid catabolism, and other processes,which were related to the abnormal metabolic function of the liver. The efficacy of Yuejuwan against PHD was achieved mainly through the regulation of high mobility group nucleosomal-binding domain 2 (HMGN2), CALD1, and Mup7 protein expression levels and correcting the biological processes and abnormal pathways related to the pathogenesis of PHD,including muscle contraction,tight junction pathway,myocardial contraction pathway,and focal adhesion pathway. ConclusionCUMS combined with high-fat diet is reasonable in the induction of the PHD model in AopE-/- mice. Yuejuwan can correct the depression and anxiety conditions of PHD model mice,reduce the aortic plaque, and recover the abnormal blood lipid and liver function levels. Furthermore, Yuejuwan can correct abnormal biological processes and pathways of PHD model mice. The differential proteins screened throughout the experiment and the involved physiological and pathological changes are the focus of the next experiment.

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