1.Establishment of liver IL-6 reporter mouse model by hydrodynamic gene transfection
Cong MA ; Yulong ZHANG ; Qianqian ZHOU ; Man ZHAO ; Xiaohui WANG ; Xinquan JIANG ; Linsheng ZHAN
Military Medical Sciences 2017;41(4):269-272,277
Objective To construct a mouse model for real-time,noninvasive and specific monitoring of inflammation activation in hepatic tissues.Methods An inflammation reporter gene was targeted to the liver by hydrodynamic gene delivery technology.Bioluminescence imaging was used to detect the firefly luciferase(Fluc) expression in the mouse liver after inflammatory stimulation.Besides,the relevance between the light intensity and inflammation level was also intensively investigated.Results pIL-6-Fluc was successfully delivered to the liver.The hydrodynamic gene delivery could cause a transient liver injury that could return normal in 5 to 7 days.The expression of pIL-6-Fluc could be induced by lipopolysaccharides(LPS) treatment with an about (46.80±13.35) fold increase at the peak value,which was significantly higher than that detected by ELISA [(4.09±0.96)fold].Conclusion An inflammation reporter mouse model is constructed in this study by hydrodynamic gene transfection,allowing noninvasive monitoring of inflammation activation specifically in hepatic tissues.The reporter model is capable of monitoring inflammation activation with a sensitivity higher than that of ELISA.
2.Preparation and Identification of A Peptide Monoclonal Antibody Against Human Papillomavirus 1 8E6
Qianqian KONG ; Zhenhua TANG ; Fenfen XIANG ; Yueping ZHAN ; Jian XU ; Rong WU ; Xiangdong KANG
Journal of Modern Laboratory Medicine 2016;31(3):30-32,35
Objective To prepare a peptide monoclonal antibody (McAb)against human papillomavirus 18E6 separately,and identify its specificity and pathogenicity.Metheds The advantage epitope peptide was designed and synthesized by ABCpred and Bcepred,and then used to immunize BALB/c mice after coupling with bovine serum albumin (BSA).And the McAb was prepared by hybridoma technique.HPV18E6 gene was amplified from cervical swab specimen containing HPV18 and insert-ed into expression vector pET-28a.The constructed recombinant plasmid was transformed to E.coli BL21(DE3)for expres-sion under induction of isopropyl thio-β-D-galactoside.The expressed protein was used to identified the McAb had been pre-pared.Results The hybridoma cell lines could constantly produce MAbs against HPV18E6 peptides.Sequencing proved that recombinant plasmid pET-28a-HPV18E6 was constructed correctly.Western blotting showed that the anti-HPV18E6 pep-tides antibody could specifically recognize HPV18E6.Conclusion A monoclonal antibody against the advantage epitope pep-tide of human papillomavirus 18E6 prepared could specifically recognize HPV18E6 specifically.
3.Study on the Prevention and Treatment of Postoperative Abdominal Adhesion by Inhibiting Epithelial-Mesenchymal Transition of Peritoneal Mesothelial Cells with Supplemented Buyang Huanwu Decoction
Minlin ZHENG ; Wenjiang FAN ; Yanan WANG ; Qianqian ZHAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(6):1458-1470
Objective To evaluated the efficacy of Supplemented Buyang Huanwu Decoction in preventing and treating postoperative abdominal adhesions.Methods One hundred and eight male SD rats were randomly divided into ordinary group,mold group,Sodium hyaluronate group and Supplemented Buyang Huanwu Decoction group.The animals were sacrificed 7,14 and 28 days after the operation,Visual scoring was performed,and samples were taken from the injury site for hematoxylin-eosin staining and Masson staining,and scanning electron microscope to observe the morphology of peritoneal mesothelial cells in the modeling site.Immunohistochemical method was used to observe the expression of epithelial mesenchymal transition(EMT)related cell marker proteins E-cadherin and α-SMA in the adhesive site.Results ①Efficacy of Supplemented Buyang Huanwu Decoction in the prevention and treatment of postoperative abdominal adhesions:The Supplemented Buyang Huanwu Decoction group can reduce the Diamod visual adhesion score(P<0.05),as well as the adhesion levels of HE staining and Masson staining(P<0.05).②Degree of damage to peritoneal mesothelial cells:Scanning electron microscopy showed that in the field of view of the Supplemented Buyang Huanwu Decoction group,paving like peritoneal mesothelial cells could be seen,indicating that the Supplemented Buyang Huanwu Decoction can improve the damage of peritoneal mesothelial cells on the serosal side of the cecum in PAA.③Expression of Epithelial mesenchymal transition-related indicators:The expression of E-Cadherin protein in the Supplemented BuyangHuanwu Decoction group increased(P<0.05).The expression of α-SMA protein in the Supplemented Buyang Huanwu Decoction group decreased(P<0.05).Conclusion The effect of Supplemented Buyang Huanwu Decoction in the prevention and treatment of postoperative abdominal adhesions is exact,which mechanism may be by reducing Epithelial mesenchymal transition of peritoneal mesothelial cells,reducing the deposition of extracellular matrix,thereby reducing peritoneal adhesion.
4.Refractive status of children and adolescents in Gaoxin District of Chengdu
Chinese Journal of School Health 2022;43(10):1565-1569
Objective:
To investigate the refractive status of children and adolescents aged 4-18 years in Chengdu, and to analyze the epidemiological characteristics of refractive parameters.
Methods:
A cross sectional survey was conducted among 82 024 children and adolescents aged 4-18 years in Gaoxin District of Chengdu from August to September 2021. The prevalence of screening myopia, low vision rate, high myopia rate, refractive status and axial development were analyzed.
Results:
The prevalence of screening myopia in students aged 4-18 years was 40.42%(33 158/82 024). Low myopia was 24.51%(20 108/82 024), moderate myopia was 13.05%(10 703/82 024) and high myopia was 2.86%(2 347/82 024). The prevalence of screening myopia was 1.81% (233/12 848) in kindergarten, 34.44%(17 095/49 644) in primary school, 79.73%(9 738/12 214) in junior high school and 83.25% (6 092/7 318) in senior high school. The rate of visual impairment increased by year from the age of 4, and the rate of myopia increased most rapidly from 6 to 15 years old, the prevalenct of high myopia was compared between adjacent age groups: there were statistical differences between 9-15 years old were more likely to be nearsighted than boys( P <0.035 7). Significant differences in screening myopia between 5- and 7-17 year old groups,and in boys and girls.The median total diopter was 0.40 D at the age of 4 and developed to -2.90 D at the age of 18 . The diopter of girls aged 8-15 years was higher than that of boys, and there was statistical significance( Z=-2.53, -4.09, -5.67, -8.64, -5.56, -4.97, -2.52, -2.14, P <0.05). The axial length gradually increased with age, with the mean value of (22.31±0.59) mm at 4 years old and (24.91±1.00) mm at 18 years old. The mean corneal curvature did not change with age (43.19±1.47)D.
Conclusion
6-15 years old is a especially critical period for myopia development. More efforts need to be taken to decrease the prevalence of myopia before 6 years old. Prevention of the development of high myopia should start before the age of 10. The prevalence of myopia in girls is higher than that in boys, more prevention and control of myopia should focus on girls.
5.Construction and immunogenicity study of recombinant MVA vaccine expressing HIV-1 CRF01_AE gp145 gene
Qianqian ZHAN ; Danying CHEN ; Jingrong YE ; Shuangqing YU ; Ke XU ; Yi ZENG ; Xia FENG
Chinese Journal of Experimental and Clinical Virology 2015;29(3):263-265
Objective To construct recombinant MVA vaccine encoding codon-optimized HIV-1 CRF01_AE gp145 gene and evaluate its immunogenicity in mice.Methods The intracellular region of codon-modified HIV-1 CRF01_AE consensus env gene obtained in our preliminary study was removed to get gp145 gene.The modified gp145 gene was cloned into shuttle plasmid pSC11.BHK-21 cells infected with wild-type MVA in advance were transfected with the recombinant shuttle plasmid.Then the recombinant MVA vaccine expressing gp145 rMVA-AEgp145 was obtained by homologous recombination.After expression of Gp145 was confirmed by WB assay,BALB/c mice were immunized with the recombinant MVA vaccine and the cellular immune responses were evaluated.Results Recombinant MVA vaccine expressed Gp145 protein efficiently and induced high level of cellular immune responses in immunized BALB/c mice.Conclusion The recombinant MVA vaccine encoding HIV-1 CRF01_AE gp145 gene was constructed successfully.This vaccine could induce strong env-specific CTL responses in mice.
6.Expressing patterns of serum inflammatory factors in patients with gas-tric cancer with different types of tongue fur
Junfeng ZHANG ; Liwei ZHANG ; Juan WU ; Qianqian XIN ; Wei DONG ; Ruiping WANG ; Zhen ZHAN
Journal of Beijing University of Traditional Chinese Medicine 2018;41(5):405-412
Objective To investigate the expressing patterns of serum inflammatory factors in patients with gastric cancer(GC)with different types of tongue fur.Methods GC patients with different types of longue fur(n=134)and non-GC subjects with different types of tongue fur(n=93)were chosen from clinic.The levels of serum inflammatory factors were detected by using electro-chemiluminescence immu-noassay(ECLIA),and correlation between types of tongue fur and serum inflammatory factors was ana-lyzed by using multi-factor association method.Results The levels of serum interleukin-17α(IL-17α) had significant difference in GC patients with different types of longue fur(P<0.05).The analysis on fur color showed that level of serum IL-17αwas 3.062 ng/L in GC patients with yellow fur and 3.982 ng/L in those with white fur(P<0.05).ROC analysis showed that the sensitivity was 72.7%,specific-ity was 91.3%and AUC(95% CI)was 0.639(0.539 -0.739)of serum IL-17αin distinguishing white fur and yellow fur,and cutoff value was 6.033 ng/L.The similar tendency was identified in non-GC subjects.The correlation analysis showed that there were 20 serum inflammatory factors with signifi-cant positive correlation in GC patients with the same types of tongue fur,and significant negative correla-tion in GC patients with different types of tongue fur.Conclusion The level of IL-17αcan significantly distinguish white tongue fur and yellow tongue fur.The white-yellow tongue fur and thin-thick tongue fur may reflect adverse inflammatory status in GC patients.
7.Identification of the Antioxidant of Chrysanthemum morifolium Using Entropy-weighted TOPSIS Method Combined with Spectrum-effect Analysis
Qianqian ZHU ; Shiqin WANG ; Xuxiang ZHOU ; Ruili NIE ; Jianting ZHAN ; Bo WANG ; Jing NIE ; Xiaochuan YE ; Dan LIU
Herald of Medicine 2024;43(6):956-964
Objective To evaluate the antioxidant function of Chrysanthemum morifolium from different origins and to identify their antioxidant material basis.Methods The HPLC fingerprints of the water extracts of C.morifolium from different origins were established.The antioxidant activities of C.morifolium were assayed by measuring the 2.2-diphenyl-l-picrylhydrazyl(DPPH),hydroxyl radical,ABTS,superoxide anion radical scavenging capacity and ferric ion reducing capacity FRAP.Entropy-weighted TOPSIS was used to calculate the weighting coefficients of the single indexes.Grey relational analysis(GRA)and partial least squares were used for spectrum-effect analysis to identify the antioxidant material basis of C.morifolium.Results A total of 16 common peaks were discovered in the fingerprint of the water extracts of 10 batches of C.morifolium,among which 13 common components were identified.All the C.morifolium samples had good antioxidant capacity,and the results of entropy-weighted TOPSIS analysis showed that the ranking of total antioxidant potency of 10 batches of C.morifolium was follows:S1>S8>S3>S5>S4>S10>S7>S2>S9>S6.The peaks of 1-5,9,10,12,14 were positively correlated with the antioxidant activity and the variable influence on projection(VIP)values were greater than 1.The correlation coefficients of these nine peaks in GRA were all greater than 0.7.Conclusion The entropy-weighted TOPSIS method combined with the spectrum-effect analysis could be used to screen out the antioxidant material basis of C.morifolium and the results provide a basis for establishing quality assessment system for C.morifolium based on Quality-markers thus improving the quality control level.
8.Effect of baotaiyin on IL-23 /Th17 immune inflammatory axis in mouse model of spontaneous abortion.
Xingxiu ZHAN ; Lijuan JIANG ; Hongping NIU ; Lijuan YANG ; Qianqian WAN ; Yanping QIAN
Journal of Central South University(Medical Sciences) 2022;47(11):1532-1539
OBJECTIVES:
The mechanism for traditional Chinese medicine in treating of recurrent spontaneous abortion is not clear. This study aims to explore the mechanism of baotaiyin in the treatment of recurrent abortion by regulating the immune inflammatory axis of interleukin (IL)-23/helper T cell (Th)17.
METHODS:
Spontaneous abortion model mice were randomly divided into a model group, 3 dose (low, medium, and high) groups of baotaiyin, with 10 mice in each group. After 14 days of medication, the levels of IL-17, IL-23, IL-10, and TGF-β in serum were detected with enzyme-linked immunosorbent assay. The proportion of Th17 and regulatory T cells (Treg) cells in spleen lymphocytes was tested with flow cytometry. The expressions of (retinoid-related orphan receptor γt, ROR-γt) and forkhead box P3 (FOXP3) mRNA in decidua tissues was detected with RT-PCR. Embryo absorption rate was counted.
RESULTS:
Compared with the model group, the absorption rate of embryo and Th17/Treg cell ratio in baotaiyin medium- and high-dose groups were decreased significantly (all P<0.05); the levels of IL-17 and IL-23 in serum were decreased (both P<0.05), while the levels of TGF-β and IL-10 in baotaiyin medium- and high-dose groups were increased (P<0.05, P<0.01, respectively); the expression of ROR-γt mRNA was decreased and the expression of FOXP3 mRNA was increased (all P<0.01) in decidua tissues of baotaiyin medium- and high-dose groups.
CONCLUSIONS
Baotaiyin inhibits the positive feedback cycle of IL-23/Th17 immune inflammatory axis, which regulates Th17/Treg cell balance, mediates the maternal and fetal immune tolerance, and prevents the recurrent abortion.
Mice
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Animals
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Female
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Humans
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Pregnancy
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Interleukin-23
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Interleukin-17/genetics*
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Interleukin-10
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Abortion, Habitual
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Transforming Growth Factor beta/genetics*
9.Composition changes and the characterization of “fried charcoal and saving properties”based on the carbon-frying process of Sophora japonica and its bud
Chunmeng XU ; Zhan LIU ; Qianqian ZHANG ; Qi LU ; Yanpeng DAI ; Xuelan ZHANG ; Dianhua SHI
China Pharmacy 2024;35(21):2622-2628
OBJECTIVE To explore the changes in chemical compositions and the characteristics of “fried charcoal and saving properties” based on the carbon-frying process of Sophora japonica and its bud. METHODS The slightly, moderately and heavily carbon-fried samples of S. japonica and its bud were prepared. The fingerprints of S. japonica, its bud and carbon-fried samples were established, and common peaks were identified. HPLC method was used to determine the contents of identified components and differential analysis was also performed; the differential components were screened by using chemometric methods, and their content ratios were used to characterize the “fried charcoal and saving properties” of S. japonica and its bud. RESULTS There were 9 common peaks in the fingerprints of S. japonica and its carbon-fried samples, 8 common peaks in those of S. japonica bud and its carbon-fried samples. In the fingerprints of S. japonica and its bud, and their different fried products, 6 components were identified, such as rutin, kaempferol-3-O-rutinoside, isorhamnetin-3-O-rutinoside, quercetin, kaempferol,isorhamnetin. Among them, the contents of rutin, kaempferol-3-O-rutinoside and isorhamnetin-3-O-rutinoside were the highest in slightly carbon-fried samples of S. japonica bud, and the contents of quercetin and isorhamnetin were the highest in moderately carbon-fried samples of S. japonica bud, and the contents of kaempferol were higher in moderately and healily carbon-fried samples of S. japonica bud. The chemometric results showed that the variable importance projection values of rutin and quercetin were both greater than 1. The range of rutin-quercetin content ratio between 9.00-14.00 and 3.00-6.00 respectively could characterize “fried charcoal and saving properties” of S. japonica and its bud. CONCLUSIONS There are significant differences in the chemical compositions of raw and fried products of S. japonica and its bud. Rutin and quercetin may be the differential components that affect their quality, and the ratio range of the two can be used to characterize the “fried charcoal and saving properties” of S. japonica and its bud in the future.
10.Isogenic human pluripotent stem cell disease models reveal ABRA deficiency underlies cTnT mutation-induced familial dilated cardiomyopathy.
Bin LI ; Yongkun ZHAN ; Qianqian LIANG ; Chen XU ; Xinyan ZHOU ; Huanhuan CAI ; Yufan ZHENG ; Yifan GUO ; Lei WANG ; Wenqing QIU ; Baiping CUI ; Chao LU ; Ruizhe QIAN ; Ping ZHOU ; Haiyan CHEN ; Yun LIU ; Sifeng CHEN ; Xiaobo LI ; Ning SUN
Protein & Cell 2022;13(1):65-71