1.iTRAQ-based proteomics reveals the mechanism of action of Yinlai decoction in treating pneumonia in mice consuming a high-calorie diet
Qianqian Li ; Tiegang Liu ; Chen Bai ; Xueyan Ma ; Hui Liu ; Zi ; an Zheng ; Yuxiang Wan ; He Yu ; Yuling Ma ; Xiaohong Gu
Journal of Traditional Chinese Medical Sciences 2024;11(1):21-32
Objective:
To uncover the underlying mechanisms of action of the Yinlai decoction on high-calorie diet-induced pneumonia through proteomics analysis.
Methods:
Based on the Gene Expression Omnibus (GEO) database, lung tissue samples from normal and high-fat diet (HFD) fed mice in the GSE16377 dataset were selected as test cohorts to identify differentially expressed genes and conduct bioinformatics analyses. In the animal experiments, mice were randomly divided into the control (N), high-calorie diet pneumonia (M), and Yinlai decoction treatment (Y) groups. Mice in the M group received high-calorie feed and a 0.5 mg/mL lipopolysaccharide solution spray for 30 min for 3 d. The mice in the Y group were intragastrically administered 2 mL/10 g Yinlai decoction twice daily for 3 d. Pathological evaluation of the lung tissue was performed. Differentially expressed proteins (DEPs) in the lung tissue were identified using quantitative proteomics and bioinformatics analyses. The drug-target relationships between Yinlai decoction and core DEPs in the lung tissue were verified using AutoDock Vina and Molecular Graphics Laboratory (MGL) Tools. DEPs were verified by western blot.
Results:
GEO data mining showed that an HFD altered oxidative phosphorylation in mouse lung tissue. The Yinlai decoction alleviated pathological damage to lung tissue and pneumonia in mice that were fed a high-calorie diet. A total of 47 DEPs were identified between the Y and M groups. Enrichment analysis revealed their association with energy metabolism pathways such as the tricarboxylic acid cycle (TCA) and oxidative phosphorylation. The protein-protein interaction network revealed that Atp5a1, Pdha1, and Sdha were the target proteins mediating the therapeutic effects of Yinlai decoction. Molecular docking results suggested that the mechanism of the therapeutic effect of Yinlai decoction involves the binding of brassinolide, praeruptorin B, chrysoeriol, and other components in Yinlai decoction to Atp5a1.
Conclusion
The Yinlai decoction alleviated lung tissue damage and pneumonia in mice that were fed a high-calorie diet by regulating the TCA and oxidative phosphorylation. Our study highlights the importance of a healthy diet for patients with pneumonia and provides a scientific basis for the prevention and treatment of pneumonia through dietary adjustments.
2.Application study on the prevention and treatment of spleen aminopeptide oral solution on seasonal allergic rhinitis
Haiyun SHI ; Kaili ZHENG ; Change FAN ; Huijiao CAI ; Tingting MA ; Huiyu NING ; Hongtian WANG ; Junjing ZHANG ; Xueyan WANG
Chinese Journal of Preventive Medicine 2024;58(8):1219-1229
Methods:A total of 392 patients with seasonal allergic rhinitis were selected from the population of the epidemiological investigation project of allergic diseases in Hohhot, Inner Mongolia. The project was led by Department of Allergy, Beijing Shijitan Hospital, Capital Medical University, and assisted by Hohhot First Hospital from April to May 2023. The patients were randomly divided into a spleen aminopeptide group (296 cases) and control group (96 cases) at a ratio of 3∶1, and the enrollment period was from June 1 to 14, 2023. The treatment group was treated with spleen aminopeptide oral solution for 12 weeks starting from 4-6 weeks (±7 days) before the pollen dispersal period, while the control group was treated with a simulated agent of spleen aminopeptide oral solution. Both the treatment group and the control group were treated with oral antihistamines and/or nasal glucocorticoids as needed during the pollen dispersal period. Evaluate the therapeutic effect by comparing the symptom scores, drug scores and quality of life scores of the two groups, and detect the expression levels of cytokines in serum. Symptom scores, quality of life scores, drug scores and laboratory results were compared by independent sample t test/Kruskal-Wallis test and χ2 test/Fisher′s exact test. Results:Compared with the control group, spleen aminopeptide treatment for 12 weeks significantly improved symptoms of nasal congestion [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=6.308, P<0.05], nasal itching [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=4.966, P<0.05], sneezing [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=5.245, P<0.05], runny nose [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=5.41, P<0.05] and tearing [ M( Q1, Q3):1(0, 2) vs. 1(0, 3), H=4.664, P<0.05]. At 12 weeks of treatment, the scores of nasal symptoms and ocular symptoms in control group and experimental group were significantly increased compared with baseline ( P<0.05). In experimental group, nasal congestion [ M( Q1, Q3):1(0, 1) vs. 1(0, 2), H=4.042, P<0.05], eye itching/foreign body sensation/redness symptom scores [ M( Q1, Q3):1(0, 2) vs. 1(0, 2), H=5.302, P<0.05] and total scores [ M( Q1, Q3):4(-1, 9) vs. 5(0, 12.5), H=3.958, P<0.05] were significantly increased. The antihistamine drug score of the splenic peptide treatment group at 6 weeks were lower than that of the control group ( H=4.232, P<0.05). After 12 weeks of treatment, the antihistamine drug score [ M( Q1, Q3):10(0, 24) vs. 19(2, 36.5), H=6.67, P<0.05] and the total drug score [ M( Q1, Q3):28.5(5, 77.5) vs. 46(6, 155.5), H=3.995, P<0.05] were significantly lower than those of the control group. The serum IL-17A levels of the treatment group were significantly lower than those of the control group after 6 weeks (0.7±1.77 vs. 0.85±1.67 ,H=10.08, P<0.05) and 12 weeks (0.81±1.63 vs. 0.94±1.73, H=5.196, P<0.05) of splenic aminopeptide treatment. Conclusions:Early treatment with spleen aminopeptide oral solution can significantly improve nasal and ocular symptoms of patients with seasonal allergic rhinitis, reduce the use of drugs during the onset period, and improve the quality of life. It may exert an immunomodulatory effect by reducing the expression level of IL-17A in the serum of patients.Objects:To conduct a study on the prevention and treatment of seasonal allergic rhinitis in Hohhot, Inner Mongolia, evaluate the preventive and therapeutic effects of spleen aminopeptide oral solution on seasonal allergic rhinitis, and explore its related mechanisms.
3.Application study on the prevention and treatment of spleen aminopeptide oral solution on seasonal allergic rhinitis
Haiyun SHI ; Kaili ZHENG ; Change FAN ; Huijiao CAI ; Tingting MA ; Huiyu NING ; Hongtian WANG ; Junjing ZHANG ; Xueyan WANG
Chinese Journal of Preventive Medicine 2024;58(8):1219-1229
Methods:A total of 392 patients with seasonal allergic rhinitis were selected from the population of the epidemiological investigation project of allergic diseases in Hohhot, Inner Mongolia. The project was led by Department of Allergy, Beijing Shijitan Hospital, Capital Medical University, and assisted by Hohhot First Hospital from April to May 2023. The patients were randomly divided into a spleen aminopeptide group (296 cases) and control group (96 cases) at a ratio of 3∶1, and the enrollment period was from June 1 to 14, 2023. The treatment group was treated with spleen aminopeptide oral solution for 12 weeks starting from 4-6 weeks (±7 days) before the pollen dispersal period, while the control group was treated with a simulated agent of spleen aminopeptide oral solution. Both the treatment group and the control group were treated with oral antihistamines and/or nasal glucocorticoids as needed during the pollen dispersal period. Evaluate the therapeutic effect by comparing the symptom scores, drug scores and quality of life scores of the two groups, and detect the expression levels of cytokines in serum. Symptom scores, quality of life scores, drug scores and laboratory results were compared by independent sample t test/Kruskal-Wallis test and χ2 test/Fisher′s exact test. Results:Compared with the control group, spleen aminopeptide treatment for 12 weeks significantly improved symptoms of nasal congestion [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=6.308, P<0.05], nasal itching [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=4.966, P<0.05], sneezing [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=5.245, P<0.05], runny nose [ M( Q1, Q3):2(1, 2) vs. 2(1, 3), H=5.41, P<0.05] and tearing [ M( Q1, Q3):1(0, 2) vs. 1(0, 3), H=4.664, P<0.05]. At 12 weeks of treatment, the scores of nasal symptoms and ocular symptoms in control group and experimental group were significantly increased compared with baseline ( P<0.05). In experimental group, nasal congestion [ M( Q1, Q3):1(0, 1) vs. 1(0, 2), H=4.042, P<0.05], eye itching/foreign body sensation/redness symptom scores [ M( Q1, Q3):1(0, 2) vs. 1(0, 2), H=5.302, P<0.05] and total scores [ M( Q1, Q3):4(-1, 9) vs. 5(0, 12.5), H=3.958, P<0.05] were significantly increased. The antihistamine drug score of the splenic peptide treatment group at 6 weeks were lower than that of the control group ( H=4.232, P<0.05). After 12 weeks of treatment, the antihistamine drug score [ M( Q1, Q3):10(0, 24) vs. 19(2, 36.5), H=6.67, P<0.05] and the total drug score [ M( Q1, Q3):28.5(5, 77.5) vs. 46(6, 155.5), H=3.995, P<0.05] were significantly lower than those of the control group. The serum IL-17A levels of the treatment group were significantly lower than those of the control group after 6 weeks (0.7±1.77 vs. 0.85±1.67 ,H=10.08, P<0.05) and 12 weeks (0.81±1.63 vs. 0.94±1.73, H=5.196, P<0.05) of splenic aminopeptide treatment. Conclusions:Early treatment with spleen aminopeptide oral solution can significantly improve nasal and ocular symptoms of patients with seasonal allergic rhinitis, reduce the use of drugs during the onset period, and improve the quality of life. It may exert an immunomodulatory effect by reducing the expression level of IL-17A in the serum of patients.Objects:To conduct a study on the prevention and treatment of seasonal allergic rhinitis in Hohhot, Inner Mongolia, evaluate the preventive and therapeutic effects of spleen aminopeptide oral solution on seasonal allergic rhinitis, and explore its related mechanisms.
4.Melatonin Attenuates Mitochondrial Damage in Aristolochic AcidInduced Acute Kidney Injury
Jian SUN ; Jinjin PAN ; Qinlong LIU ; Jizhong CHENG ; Qing TANG ; Yuke JI ; Ke CHENG ; Rui WANG ; Liang LIU ; Dingyou WANG ; Na WU ; Xu ZHENG ; Junxia LI ; Xueyan ZHANG ; Zhilong ZHU ; Yanchun DING ; Feng ZHENG ; Jia LI ; Ying ZHANG ; Yuhui YUAN
Biomolecules & Therapeutics 2023;31(1):97-107
Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN).AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.
5.A highly sensitive bio-barcode immunoassay for multi-residue detection of organophosphate pesticides based on fluorescence anti-quenching
Xu LINGYUAN ; Zhang XIUYUAN ; El-Aty A.M.ABD ; Wang YUANSHANG ; Cao ZHEN ; Jia HUIYAN ; Salvador J.-PABLO ; Hacimuftuoglu AHMET ; Cui XUEYAN ; Zhang YUDAN ; Wang KUN ; She YONGXIN ; Jin FEN ; Zheng LUFEI ; Pujia BAIMA ; Wang JING ; Jin MAOJUN ; D.Hammock BRUCE
Journal of Pharmaceutical Analysis 2022;12(4):637-644
Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and rice.Gold nanoparticles were functionalized with monoclonal antibodies against the tested OPs.DNA oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal amplification.The detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the fluorophore.The resulting fluorescence signal en-ables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concen-tration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,respectively.The mean recovery ranged between 80.3%and 110.8%with relative standard deviations of 7.3%-17.6%,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.
6.Structural and functional characteristics of microbiota in oropharynx of sub-healthy children with gastrointestinal heat retention syndrome differentiated by traditional Chinese medicine
Zhen JIANHUA ; Yu HE ; Li XIAOFEI ; Dong FEI ; Zheng ZI'AN ; Ma XUEYAN ; Wan YUXIANG ; Liu TIEGANG ; Fan LU ; Gu XIAOHONG
Journal of Traditional Chinese Medical Sciences 2022;9(3):230-236
Objective:To explore the structural and functional characteristics of microbiota in oropharynx of sub-healthy children with gastrointestinal heat retention syndrome(GHRS)differentiated by traditional Chinese medicine,and screen the biometric operational taxonomic units(OTUs)to assist the clinical diagnosis.Methods:We recruited children according to the"GHRS diagnostic scale",collected their oropharyngeal swabs,and sequenced the 16S rDNA V4 region.We described the bacterial structure with alpha-indexes,beta-distances,and relative abundances;moreover,we screened the differential genera/OTUs with Wilcoxon rank-sum test,MetagenomeSeq analysis,and linear discriminant analysis effect size(LEfSe)analysis,in which biometric OTUs were selected to construct the receiver operating characteristic curve to verify the diagnostic value.The bacterial function was predicted with Kyoto Encyclopedia of Genes and Genomes pathways according to 16S rDNA gene by using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States.Results:The study population was composed of 10 children with GHRS and 10 healthy control children.GHRS children were more likely to overeat(gluttony,P=.033).Alpha-indexes,such as Sobs,abundance-based coverage estimator,Bootstrap,and Qstat,were significantly higher in the GHRS group,while beta-distances did not exhibit any significant intergroup differences.There were 9 differently distributed non-predominant genera between the groups in Wilcoxon rank-sum test,as well as 13 non-predominant genera in MetagenomeSeq analysis and 3 non-predominant OTUs in LEfSe analysis.OTU44 and OTU196 were used to construct the receiver operating characteristic curve,and the area under curve was 0.92.Predicted functions showed that pathways related to oxidative phosphorylation and carbon metabolism were enriched in healthy control samples,while the pathway related to renin secretion was remarkably enriched in GHRS samples.Conclusion:Unique oropharyngeal microbial structure and function were identified in GHRS children.OTU44 and OTU196 were specific OTUs,which could be used as biomarkers of GHRS to assist clinical diagnosis.
7.Optimization of blending method of Shuanghuanglian injection and its stability study in different solvents
Bin HUAI ; Xueyan ZHENG ; Zhi CHEN ; Jie DU
International Journal of Traditional Chinese Medicine 2021;43(7):674-679
Objective:To optimize the blending method of Shuanghuanglian injection, and to investigate its stability in different solvents (0.9% sodium chloride injection, 5% glucose injection, 10% glucose injection, glucose and sodium chloride injection). Methods:By using orthogonal test to optimize the best dissolution method of Shuanghuanglian injection By measuring the content change of insoluble particles, pH value and principal components (baicalin, forsythione, chlorogenic acid) in the finished products to investigatethe stability of Shuanghuanglian injection in different solvents. Results:The optimal blending method of Shuanghuanglian injection was to add 5 ml sterilized water for injection into the vial and oscillate at 1 200 r/min frequency for 5 min. The main constituents of Shuanghuanglian injection were stable in 8 h in the infusion of four kinds of finished products. Insoluble particles in 0.9% sodium chloride infusion and 5% glucose infusion met the requirements within 8 h, and insoluble particles in 10% glucose infusion and 6 h glucose and sodium chloride infusion met the requirements. The pH value of 0.9% sodium chloride infusion within 8 h met the optimal requirements of the best compatibility, 5% glucose infusion within 2 h met the requirements, and 4 h sodium chloride infusion met the requirements of the best compatibility. Conclusion:This study optimized the best preparation method of Shuanghuanglian (freeze-dried) for injection. Sodium chloride injection should be used as the solvent to prepare finished infusion in clinical application, and 5% glucose injection should be prepared just before use.
8. Relationship between work-related musculoskeletal disorders and ergonomic factor load in shipyard workers
Ying QU ; Xifeng CHEN ; Wei ZHANG ; Chengbin ZHENG ; Xueyan ZHANG ; Ning JIA ; Siwu ZHONG ; Qing XU ; Xi ZHANG ; Zhongxu WANG
China Occupational Medicine 2020;47(03):260-267
OBJECTIVE: To assess the effect of ergonomic factor load on work-related musculoskeletal disorders(WMSDs) in shipyard workers. METHODS: A total of 751 shipyard workers were selected as the research subjects using judgment sampling method. The exposure level of adverse ergonomic factors was assessed using the Quick Exposure Check method. The prevalence of WMSDs was investigated using the revised Musculoskeletal Disorders Investigating Questionnaire of our research group, and the relationship between them was analyzed. RESULTS: The proportions of high or very high level of work load exposure to the neck, back, shoulder and hand in shipyard workers from high to low were 66.4%, 63.5%, 59.8% and 43.7%(P<0.01) respectively. The proportions of occupational stress, driving, vibration and working rhythm at high or very high exposure level were 20.0%, 4.1%, 22.9% and 3.2%, respectively. The prevalence of WMSDs in four body parts of shipyard workers from high to low was the back, neck, hand and shoulder(the prevalence were 44.2%, 31.2%, 26.9% and 26.6%, respectively, P<0.01). After excluding the influence of confounding factors, multiple logistic regression analysis showed that the higher the labor load exposure level and longer of the vibration tool using, the higher the risk of shoulder WMSDs [odds ratio(OR) and 95% confidence interval(CI) were 1.25(1.04-1.51) and 1.33(1.05-1.69), respectively, P<0.05]. The higher the level of occupational stress, the higher the risk of back and neck WMSDs [OR(95%CI) was 1.29(1.05-1.58) and 1.42(1.15-1.77), respectively, P<0.05]. CONCLUSION: There was a dose-effect relationship between the exposure level of shoulder load, the time of using vibration tools and the shoulder WMSDs, and there was a dose-effect relationship between the occupational stress level and the WMSDs in the back and neck.
9. Effects of curcumin on pulmonary inflammation and p38 expression in an infant murine model of pneumococcal pneumonia
Wenjuan DUAN ; Song QING ; Jieling WANG ; Xueyan ZHENG ; Kai YANG
International Journal of Traditional Chinese Medicine 2019;41(10):1096-1100
Objective:
To investigate the effects of curcumin on pneumococcal pneumonia-induced pneumonia, apoptosis and p38 MAPK expression in infant mice.
Methods:
A total of 60 male infant C57BL/6 mice at three weeks of age were randomly divided into 6 groups: control group, model group, high-dose curcumin treatment group, middle-dose curcumin treatment group, low-dose curcumin treatment group and SB203580 treatment group. The Curcumin and SB203580 were intraperitoneally applied at doses of 200, 60, and 20 mg/kg (for curcumin) and 100 mg/kg (for SB203580) from two days before bacterial infection to three days post-infection. The control group and model group were intraperitoneally injected with an equal volume of saline. The model group, curcumin treatment groups and SB203580 treatment group were transnasally inoculated with approximately 106 CFU/ml of pneumococcal pneumonia in 50 μl of PBS applied to the tip of the nose to establish the experimental pneumococcal pneumonia. Subsequently, all the mice were killed and lung tissues were harvested for hematoxylin-eosin staining, calculation of lung score indexes, measurement of IL-1β and TNF-α contents by ELISA, and measurement of Bax, Bcl-2and p38 MAPK expression by Western blot.
Results:
Compared to the model group, the edema score (0.50 ± 0.10, 1.51 ± 0.16, 1.38±0.11,
10.An Enzymolysis-Assisted Agrobacterium tumefaciens-Mediated Transformation Method for the Yeast-Like Cells of Tremella fuciformis
Yuanyuan WANG ; Danyun XU ; Xueyan SUN ; Lisheng ZHENG ; Liguo CHEN ; Aimin MA
Mycobiology 2019;47(1):59-65
Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.
Agrobacterium
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Blotting, Southern
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Digestion
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Genome
;
Genomics
;
Methods
;
Microscopy, Fluorescence
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Oxidoreductases
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Polymerase Chain Reaction
;
Real-Time Polymerase Chain Reaction
;
Wounds and Injuries


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