1.Characterization of intestinal flora and transcriptomics in rats with gastric cancer
Chenxi ZHANG ; Jijuan LI ; Feicheng ZHANG ; Tianyu GAO ; Xinyue LIANG ; Lijia PAN
Acta Laboratorium Animalis Scientia Sinica 2025;33(1):70-81
Objective To analyze differences in the intestinal microbiota and transcriptomics between N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)gastric cancer rats and normal rats and to analyze the correlation between the two,so as to provide a reference for related studies using MNNG gastric cancer rats as a model.Methods A total of 12 Wistar rats were randomly divided into normal(NM)and gastric cancer(GC)groups.The GC group was given a concentration of 20 mg/mL of MNNG by gavage with a dose of 100 g/mL once a day,and the NM group was given the same amount of normal saline by gavage.Samples were collected for testing after 16 weeks of continuous intervention.The gastric tissues were collected and stained by HE staining to observe morphological changes in the gastric mucosa of the two groups,and the expression levels of differential genes were detected by transcriptome sequencing.The cecal contents were collected for 16S rRNA sequencing.Results(1)Visual observation and HE results showed that the volume of gastric mucosa in the NM group was normal,the surface was glossy,the gastric wall was elastic,the direction of the mucosal folds was regular,there were no hyperplasia or hemorrhagic spots.In the GC group,the volume of gastric mucosa was reduced,the gastric wall was thinned,elasticity was poor,the direction of the folds was disordered and irregular,and there was a bulge accompanied by yellow-black keratotic hyperplasia.In the NM group,the squamous epithelial layer,submucosa,and muscular layer of the gastric mucosa were clear,with no hyperplasia and keratinization.In the GC group,the gastric mucosa had disorganized layers and cell polarity,with different cell morphologies;the squamous epithelial layer was destroyed,and squamous epithelial cells were hyperplasic,keratinized,and had invaded the muscular layer by proliferation.The modeling was considered successful.(2)The results of intestinal microbiota sequencing showed that the abundance of Akkermansia and Lactobacillus in MNNG gastric cancer rats decreased significantly,and the abundance of the rumen coccaceae Prevonella,and Blauter increased significantly.(3)The three key pathways obtained by transcriptomic sequencing and KEGG pathway enrichment analysis were amebiasis,systemic lupus erythematosus,and the PI3K-Akt signaling pathway,and five genes differentially enriched in these three pathways were those for MCPT8I2,IGH-6,IGHG1,ACTN2,and VEGF-D.(4)Combined analysis of intestinal microbiota and transcriptomics showed that_UCG-005,Prevonella_UCG-003 and Brautella were positively correlated with amebiasis,systemic lupus erythematosus,and the PI3K-Akt signaling pathway.Conclusions The abundance of intestinal microbiota in gastric cancer rats formed by MNNG gavage is different from that of normal rats.The genes for MCPT8I2,IGH-6,IGHG1,ACTN2 and VEGF-D may be up-regulated in gastric cancer induced by MNNG gavage.Combined analysis of intestinal microbiota and differential genes suggested that the mechanism of MNNG carcinogenesis may be mainly related to the destruction of gastric mucosa and the inflammatory response.
2.Characterization of intestinal flora and transcriptomics in rats with gastric cancer
Chenxi ZHANG ; Jijuan LI ; Feicheng ZHANG ; Tianyu GAO ; Xinyue LIANG ; Lijia PAN
Acta Laboratorium Animalis Scientia Sinica 2025;33(1):70-81
Objective To analyze differences in the intestinal microbiota and transcriptomics between N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)gastric cancer rats and normal rats and to analyze the correlation between the two,so as to provide a reference for related studies using MNNG gastric cancer rats as a model.Methods A total of 12 Wistar rats were randomly divided into normal(NM)and gastric cancer(GC)groups.The GC group was given a concentration of 20 mg/mL of MNNG by gavage with a dose of 100 g/mL once a day,and the NM group was given the same amount of normal saline by gavage.Samples were collected for testing after 16 weeks of continuous intervention.The gastric tissues were collected and stained by HE staining to observe morphological changes in the gastric mucosa of the two groups,and the expression levels of differential genes were detected by transcriptome sequencing.The cecal contents were collected for 16S rRNA sequencing.Results(1)Visual observation and HE results showed that the volume of gastric mucosa in the NM group was normal,the surface was glossy,the gastric wall was elastic,the direction of the mucosal folds was regular,there were no hyperplasia or hemorrhagic spots.In the GC group,the volume of gastric mucosa was reduced,the gastric wall was thinned,elasticity was poor,the direction of the folds was disordered and irregular,and there was a bulge accompanied by yellow-black keratotic hyperplasia.In the NM group,the squamous epithelial layer,submucosa,and muscular layer of the gastric mucosa were clear,with no hyperplasia and keratinization.In the GC group,the gastric mucosa had disorganized layers and cell polarity,with different cell morphologies;the squamous epithelial layer was destroyed,and squamous epithelial cells were hyperplasic,keratinized,and had invaded the muscular layer by proliferation.The modeling was considered successful.(2)The results of intestinal microbiota sequencing showed that the abundance of Akkermansia and Lactobacillus in MNNG gastric cancer rats decreased significantly,and the abundance of the rumen coccaceae Prevonella,and Blauter increased significantly.(3)The three key pathways obtained by transcriptomic sequencing and KEGG pathway enrichment analysis were amebiasis,systemic lupus erythematosus,and the PI3K-Akt signaling pathway,and five genes differentially enriched in these three pathways were those for MCPT8I2,IGH-6,IGHG1,ACTN2,and VEGF-D.(4)Combined analysis of intestinal microbiota and transcriptomics showed that_UCG-005,Prevonella_UCG-003 and Brautella were positively correlated with amebiasis,systemic lupus erythematosus,and the PI3K-Akt signaling pathway.Conclusions The abundance of intestinal microbiota in gastric cancer rats formed by MNNG gavage is different from that of normal rats.The genes for MCPT8I2,IGH-6,IGHG1,ACTN2 and VEGF-D may be up-regulated in gastric cancer induced by MNNG gavage.Combined analysis of intestinal microbiota and differential genes suggested that the mechanism of MNNG carcinogenesis may be mainly related to the destruction of gastric mucosa and the inflammatory response.
3.Risk factors and progress in diagnosis and treatment of iliopsoas impingement after total hip replacement
Hongxun ZHU ; Jijuan LYU ; Peng ZHANG ; Zhi FENG ; Qinghe YE
Chinese Journal of Orthopaedics 2024;44(23):1567-1572
Total hip arthroplasty (THA) is a progressively refined orthopaedic surgery with excellent long-term survival rates, but it still faces problems such as postoperative pain, infection and loosening, especially groin pain. Iliopsoas impingement (IPI) is a relatively rare and overlooked cause of groin pain after THA, which often leads to delayed diagnosis and inappropriate treatment due to lack of awareness. IPI refers to the pain in the groin area caused by abnormal contact between the iliopsoas and the front of the acetabulum. There are many risk factors for IPI after THA including acetabular cup protrusion, osteophyte impingement, screw protrusion, bone cement extravasation, improper placement of acetabular prosthesis, and changes in lower limb length. The diagnosis of IPI is primarily based on physical examination, imaging findings, and a diagnostic treatment of pain relief following fluoroscopic or ultrasound-guided injection of corticosteroids and local anesthesia into the iliopsoas tendon sheath. Other potential causes such as hip dislocation, periprosthetic infection, loosening or fracture should be excluded. Treatment of IPI includes non-surgical treatment (non-steroidal anti-inflammatory drugs, physical therapy and ultrasound-guided injections of corticosteroids and local anesthesia in the iliopsoas tendon sheath), iliopsoas tenotomy, and acetabular cup revision, all three of which should be performed stepwise to maximize patient benefit.
4.Risk factors and progress in diagnosis and treatment of iliopsoas impingement after total hip replacement
Hongxun ZHU ; Jijuan LYU ; Peng ZHANG ; Zhi FENG ; Qinghe YE
Chinese Journal of Orthopaedics 2024;44(23):1567-1572
Total hip arthroplasty (THA) is a progressively refined orthopaedic surgery with excellent long-term survival rates, but it still faces problems such as postoperative pain, infection and loosening, especially groin pain. Iliopsoas impingement (IPI) is a relatively rare and overlooked cause of groin pain after THA, which often leads to delayed diagnosis and inappropriate treatment due to lack of awareness. IPI refers to the pain in the groin area caused by abnormal contact between the iliopsoas and the front of the acetabulum. There are many risk factors for IPI after THA including acetabular cup protrusion, osteophyte impingement, screw protrusion, bone cement extravasation, improper placement of acetabular prosthesis, and changes in lower limb length. The diagnosis of IPI is primarily based on physical examination, imaging findings, and a diagnostic treatment of pain relief following fluoroscopic or ultrasound-guided injection of corticosteroids and local anesthesia into the iliopsoas tendon sheath. Other potential causes such as hip dislocation, periprosthetic infection, loosening or fracture should be excluded. Treatment of IPI includes non-surgical treatment (non-steroidal anti-inflammatory drugs, physical therapy and ultrasound-guided injections of corticosteroids and local anesthesia in the iliopsoas tendon sheath), iliopsoas tenotomy, and acetabular cup revision, all three of which should be performed stepwise to maximize patient benefit.
5.Clustering of health-risk behaviors among children and adolescents in Yancheng City
LU Xiaomin ; ZHAO Jijuan ; SUN Zhongyou ; ZHANG Xueyan ; CHEN Shenglan
Journal of Preventive Medicine 2023;35(4):331-334
Objective:
To investigate the clustering of health-risk behaviors and its influencing factors among children and adolescents in Yancheng City, Jiangsu Province, so as to provide insights into the prevention and control of health-risk behaviors among children and adolescents.
Methods:
Students were randomly sampled from 4 primary schools, 4 junior high schools and 4 senior high schools in Yancheng City using a multi-stage stratified cluster random sampling method from September to December 2021. Students' demographics and 12 health-risk factors including unhealthy diet, insufficient physical activity and attempted smoking were collected using the Student's Health Status and Influencing Factors Questionnaire by Jiangsu Provincial Center for Disease Control and Prevention, and factors affecting the clustering of health-risk behaviors were identified using a multivariable linear regression model.
Results:
A total of 2 925 valid questionnaires were recovered, and the respondents included 1 611 boys (55.08%) and 1 314 girls (44.92%). A total of 2 896 respondents were detected with health-risk behaviors, with a detection rate of 99.09%, and 2 772 respondents were detected with clustering of health-risk behaviors (93.06%). Insufficient sleep, insufficient physical activity and insufficient duration of outdoor activity were predominant patterns of clustering. The median number of health-risk behaviors was 4.00 (interquartile range, 2.00) per capita. Multivariable linear regression analysis showed that boys (β=0.232), grade (junior high school, β=0.519; senior high school, β=0.427), urban area (β=0.241), living at school (β=0.395), family structure (single parental family, β=0.188; other families, β=0.344) and father's education level of primary school and below (β=0.369) were factors affecting clustering of health-risk behavior among primary and high school students.
Conclusions
The detection of health-risk behaviors is high among children and adolescents in Yancheng City, and insufficient sleep, insufficient physical activity and insufficient duration of outdoor activity are predominant health-risk behaviors. Boys, junior high school and above, urban areas, living at schools, single parents, and fathers with a low educational level lead to a high degree of clustering of health-risk behaviors.
6.Analysis of volatile organic compounds in exhaled breath after radiotherapy.
Dianlong GE ; Xue ZOU ; Yajing CHU ; Jijuan ZHOU ; Wei XU ; Yue LIU ; Qiangling ZHANG ; Yan LU ; Lei XIA ; Aiyue LI ; Chaoqun HUANG ; Pei WANG ; Chengyin SHEN ; Yannan CHU
Journal of Zhejiang University. Science. B 2022;23(2):153-157
Radiotherapy uses high-energy X-rays or other particles to destroy cancer cells and medical practitioners have used this approach extensively for cancer treatment (Hachadorian et al., 2020). However, it is accompanied by risks because it seriously harms normal cells while killing cancer cells. The side effects can lower cancer patients' quality of life and are very unpredictable due to individual differences (Bentzen, 2006). Therefore, it is essential to assess a patient's body damage after radiotherapy to formulate an individualized recovery treatment plan. Exhaled volatile organic compounds (VOCs) can be changed by radiotherapy and thus used for medical diagnosis (Vaks et al., 2012). During treatment, high-energy X-rays can induce apoptosis; meanwhile, cell membranes are damaged due to lipid peroxidation, converting unsaturated fatty acids into volatile metabolites (Losada-Barreiro and Bravo-Díaz, 2017). At the same time, radiotherapy oxidizes water, resulting in reactive oxygen species (ROS) that can increase the epithelial permeability of pulmonary alveoli, enabling the respiratory system to exhale volatile metabolites (Davidovich et al., 2013; Popa et al., 2020). These exhaled VOCs can be used to monitor body damage caused by radiotherapy.
Breath Tests/methods*
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Exhalation
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Humans
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Quality of Life
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Respiratory System/chemistry*
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Volatile Organic Compounds/analysis*
7.Purification, characterization and partial primary structure analysis of rutin-degrading enzyme in tartary buckwheat seeds.
Yuwei ZHANG ; Jie LI ; Yong YUAN ; Jijuan GU ; Peng CHEN
Chinese Journal of Biotechnology 2017;33(5):796-807
Rutin-degrading enzymes (RDE) can degrade rutin into poorly water soluble compound, quercetin, and cause the bitter taste in tartary buckwheat. In the present study RDE from Yu 6-21 tartary buckwheat seeds was purified by ammonium sulphate precipitation, followed by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B, ion exchange chromatography on CM-Cellulose and gel filtration chromatography on Sephadex G-150. Purified RDE showed single band with molecular weight of 66 kDa on SDS-PAGE. The optimum pH and temperature of RDE were 5.0 and 50 ℃ respectively. The Km was 0.27 mmol/L, and the Vmax was 39.68 U/mg. The RDE activity could be inhibited by Cu²⁺, Zn²⁺, Mn²⁺ and EDTA, and showed tolerance to 50% methanol (V/V). The N terminal sequence (TVSRSSFPDGFLFGL) was obtained by Edman degradation method and 15 internal peptide sequences were determined by MALDI-TOF-MS (matrix-assisted laser desorption ionization time of flight mass spectrometry). These results established the foundations for identification of the candidate gene of RDE via transcriptome data and further studying RDE biological function.
8.Comparison of antibody persistence after primary immunization with 5 μg and 10 μg recombinant hepatitis B vaccine among newborns with normal and high response: a five-year following-up
Li ZHANG ; Wei ZHANG ; Jingjing LYU ; Jijuan ZHANG ; Jiaye LIU ; Bingyu YAN ; Yi FENG ; Xiaofeng LIANG ; Fuqiang CUI ; Fuzhen WANG ; Guomin ZHANG ; Aiqiang XU
Chinese Journal of Epidemiology 2017;38(9):1156-1160
Objective To compare the antibody persistence 5 years after primary immunization with 5 μg and 10 μg recombinant hepatitis B vaccine (HepB) among newborns with normal and high response.Methods Newborns who completed three doses of 5 μ g HepB made by recombinant dexyribonucleic acid technique in Saccharomyces (HepB-SC) or 10 μg HepB made by recombinant dexyribonucleic acid technique in Hansenula polymorpha (HepB-HP) were recruited.Standardized questionnaire was used and blood samples were collected 1-6 months (T0) and five years (T1) after the third dose respectively.The titer of anti-HBs was detected by chemiluminescence microparticle imunoassay (CMIA).Those who achieved normal or high antibody response (anti-HBs titer ≥100 mIU/ml) were included in the study and the positive rate (≥ 10 mIU/ml) and titer of anti-HBs at T1 were compared between 5 μg HepB group and 10 μg HepB group.Multivariable analysis was conducted to identify the independent factors associated with the antibody persistence.Results The positive rate of anti-HBs at T1 was 49.92% (943/1 883) and 75.92% (1 135/1 495) respectively in 5 μg HepB group and 10 μg HepB group,the difference was significant (x2=237.75,P<0.001).The anti-IBs geometric mean concentrations at T1 were 10.23 mIU/ml (95%CI:9.38-11.16) and 28.91 mIU/ml (95%CI:26.65-31.35) in the two groups respectively,the difference was also significant (F=280.36,P<0.001).Among those whose anti-HBs titer was < 10 mIU/ml at T1,the distributions of anti-HBs titer were significantly different between 5 μg HepB group and 10 μg HepB group (x2=39.75,P< 0.001).The multivariable analysis showed that dosage of HepB was independently associated with both positive rate and titer of anti-HBs at T1 after excluding the other factors [P<0.001,OR=1.44(95% CI:1.20-1.73);P<0.001,β =0.27 (95% CI:0.14-0.40)].Conclusion Five year anti-HBs persistence after primary immunization with 10 μg HepB might be better than that after primary immunization with 5 μg HepB among infants who achieved normal or high anti-HBs response after primary HepB immunization.
9.Detection and identification of viable Listeria monocytogenes by real-time PCR
Dazhi JIN ; Jijuan CAO ; Zheng ZHANG ; Mingjie XIE ; Shuirong ZHU
Chinese Journal of Microbiology and Immunology 2008;28(10):941-945
Objective To establish a rapid,sensitive and specific assay based on real-time PCR combined with reverse transcription for detecting and identifying viable Listeria monocytogenes.Methods The hlyA gene of Listeria monocytogenes was chosen as target,and then the primers and TaqMan probe were designed.Both ends of probe were modified with two different fluorescence groups.The PCR reaction was optimized systematically.The mRNA of Listeria monocytogenes was extracted,and then reverse transcription was performed through random primer.The cDNA Was detected by real-time PCR.Then the specificity,sensitivity and reproducibility of real-time PCR were estimated.In final,real-time PCR was applied to detect 20 mocked double-blind samplea.Results Viable Listeria monocytogenes were detected by real-time PCR accurately and quickly,and meanwhile,none of other bacteria and non-viable Listeria monocytogenes could be identified.The sensitivity was 10 CFU/ml in pure culture and 103CFU/ml for mocked samples respectively.The coefficient of variation of intra-assay and inter-assay Was less than 5%.When this assay was applied directly to identify 20 mocked double-blind samples,10 of these were positive to viable Listeria monocytogenes,5 were negative to non-viable Listeria monocytogenes,and 5 were negative to other pathogens.Conclusion It is demonstrated that real-time PCR is a reliable,accurate and feasible assay for viable Listeria monocytogenes.The establishment of this assay provided complete data for analysis and diagnosis in the field of food safety and epidemiologic survey.


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