1.Newcastle disease virus suppresses antigen presentation via inhibiting IL-12 expression in dendritic cells
NAN FULONG ; NAN WENLONG ; YAN XIN ; WANG HUI ; JIANG SHASHA ; ZHANG SHUYUN ; YU ZHONGJIE ; ZHANG XIANJUAN ; LIU FENGJUN ; LI JUN ; ZHOU XIAOQIONG ; NIU DELEI ; LI YIQUAN ; WANG WEI ; SHI NING ; JIN NINGYI ; XIE CHANGZHAN ; CUI XIAONI ; ZHANG HE ; WANG BIN ; LU HUIJUN
Journal of Zhejiang University. Science. B 2024;25(3):254-270,后插1-后插4
As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.
2.Aqueous extract of Fritillaria ussuriensis attenuates nonalcoholic fatty liver disease in mice by activating AMPK/ACC pathway and regulating intestinal flora
Shimin XIE ; Yue LI ; Zhaopeng ZHANG ; Xia YANG ; Yiquan LI ; Jicheng HAN ; Yining WAN ; Huidan CHEN ; Ningyi JIN ; Yilong ZHU ; Guangze ZHU
Chinese Journal of Pathophysiology 2024;40(11):2090-2098
AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.
3.Eukaryotic expression,purification and immunoefficacy evaluation of ferritin nan-oparticles of dengue virus type Ⅱ
Junduo BAO ; Xiangshu QIU ; Yan GAO ; Jiaqi ZHANG ; Xiao LI ; Xin JIN ; Huijun LU ; Ningyi JIN
Chinese Journal of Veterinary Science 2024;44(6):1120-1126
The E protein of dengue virus type Ⅱ was presented on ferritin of Helicobacter pylori to construct a novel dengue nanoparticle vaccine candidate,and the immunological indexes of the vac-cine were evaluated,aiming to provide new ideas for the development of dengue vaccine.The re-combinant plasmid of E-Ferritin was optimized and synthesized,and then transfected into HEK-293F cells.The recombinant protein was expressed,identified,purified and analyzed.Mice were im-munized with E-Ferritin nanoparticle vaccine by intramuscular injection on the hind limbs on the day 0,14 and 28.ELISA,neutralization test,flow cytometry and lymphocyte proliferation test were used to detect the levels of specific antibodies,neutralizing antibodies,CD3+,CD4+and CD8+T lymphocytes in spleen cells and the proliferation of spleen lymphocytes after specific stimulation.The target protein with a size of about 69 kDa was expressed in the cells with a single band.The purified protein concentration was 0.407 g/L,and the purity was 82.32%.The results from transmission electron microscopy showed that E-Ferritin protein could be recombined into a particle structure with a particle size of about 50 nm.The results of mouse immune experiments showed that E-Ferritin protein had good immunogenicity.The average specific antibody titer of E-ferritin protein in serum was 1∶92 160 after immunization 42 d.The main subclass of antibody was IgGl.The results of flow cytometry showed that E-Ferritin as an immunogen could induce higher levels of CD4+and CD8+T lymphocyte immune response.In lymphocyte proliferation test,the level of specific stimulation in the vaccine group was significantly higher than that in the non-specific stimulation group.In conclusion,the dengue virus envelope protein ferritin nanoparticle vaccine constructed in this study has good immunogenicity,which can provide reference for the de-velopment of new dengue vaccine candidates.
4.Detection of Japanese encephalitis virus in livestock in Xinjiang Uygur Autono-mous Region of China from 2021 to 2022
Bing LI ; Yan GAO ; Xinyu CAO ; Xiangshu QIU ; Aijian QIN ; He ZHANG ; Ningyi JIN
Chinese Journal of Veterinary Science 2024;44(8):1713-1718
The SYBR Green Ⅰ real-time fluorescence quantitative PCR detection method was used to determine the prevalence of Japanese encephalitis virus(JEV)in mosquito vectors and cattle se-rum samples in Xinjiang.The E gene fragment of the JEV strain was amplified by PCR,cloned into a pEASY-Blunt vector,produced as a recombinant plasmid,and its sensitivity,specificity and re-producibility were verified.Between 2021 and 2022,serum samples were taken in the regions of Hami,Altay,Ili,Aksu,and Kashi in order to monitor the prevalence of JEV in livestock in Xin-jiang.The positive rate was discovered and evaluated using the established detection method.The established detection method showed a good linear relationship,and the detection interval was 4.03X102-4.03×109 copies/pL.The correlation coefficient was 0.995,the slope was-3.431,and the extreme value of the lower limit of sensitivity was 4.03 × 102 copies/pL.This method has no specific amplification for Zika virus(ZIKV)and Dengue virus(DENV).The intra group coefficient of variation of reproducibility was 0.53%-1.27%,and the inter group coefficient of variation was 0.48%-1.43%.Using this method to detect serum samples from livestock in Xinjiang from 2021 to 2022,the total positive rate was 3.28%,with positive detection rates in horses,cows,and sheep being 2.35%,6.77%,and 3.74%respectively,the virus was identified as Type Ⅰ JEV.A SYBR Green Ⅰ real-time PCR method for the detection of genotype 1 JEV was established.JEV was de-tected in the serum of horses,cattle and sheep in Xinjiang,with a total positive rate of 3.11%.
5.Difference analysis of 18F-FMISO PET/CT hypoxia imaging in response to heavy ion radiotherapy in patients with non-small cell lung cancer
Mingyu LIU ; Ningyi MA ; Jian CHEN ; Caiyue REN ; Fuquan ZHANG ; Jingfang MAO ; Kailiang WU ; Guoliang JIANG ; Yun SUN ; Shaoli SONG ; Jingyi CHENG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2023;43(1):11-15
Objective:To explore the clinical value of 18F-fluoromisonidazole (FMISO) PET/CT hypoxia imaging in early response to heavy ion radiotherapy in patients with non-small cell lung cancer(NSCLC). Methods:From April 2018 to January 2021, the 18F-FMISO PET/CT images of 23 NSCLC patients (19 males, 4 females; age (64.9±10.3) years) who received heavy ion radiotherapy in Shanghai Proton and Heavy Ion Center were retrospectively analyzed. The evaluation parameters included tumor volume (TV), tumor to background ratio (TBR) before and after radiotherapy. Patients were divided into hypoxia group and non-hypoxia group with the baseline TBR value≥1.4 as hypoxia threshold. Wilcoxon signed rank test was used to compare the differences of TV and TBR before and after radiotherapy in 2 groups. Results:Of 23 NSCLC patients, 17 were hypoxia and 6 were non-hypoxia. Compared with the baseline, TV after the radiotherapy (59.44(22.86, 99.43) and 33.78(8.68, 54.44) cm 3; z=-3.05, P=0.002) and TBR after the radiotherapy (2.25(2.09, 2.82) and 1.42(1.24, 1.67); z=-3.39, P=0.001) of the hypoxia group were significantly lower, while TV (16.19(6.74, 36.52) and 8.59(4.38, 25.47) cm 3; z=-1.57, P=0.120) and TBR (1.19(1.05, 1.27) and 1.10 (0.97, 1.14); z=-1.89, P=0.060) of the non-hypoxia group decreased with no significant differences. Conclusions:Hypoxic NSCLC tumors are sensitive to heavy ion radiation. Compared with non-hypoxic tumors, hypoxic tumors respond more quickly, and a significant reduction in TV can be observed early after radiotherapy. Heavy ion radiation can significantly improve tumor hypoxia.
6.Mechanism of action of exosomes in the development and progression of hepatitis B virus-related chronic hepatitis and hepatocellular carcinoma
Ziwei GUO ; Hening CHEN ; Xu CAO ; Jiaxin ZHANG ; Ningyi ZHANG ; Qian JIN ; Xiaoke LI ; Yongan YE
Journal of Clinical Hepatology 2022;38(9):2125-2129
Exosomes (EXOs) are formed by intracellular multivesicular bodies and carry a variety of biomacromolecules such as lipids, proteins, encoding and non-coding RNAs, and mitochondrial DNA. EXOs can be released in vivo by different cell types, including hepatocytes, hepatic stellate cells, and immune cells and play the role of intercellular communication. More and more studies have shown that EXOs are involved in the development, progression, and prognosis of chronic hepatitis B (CHB) and hepatocellular carcinoma (HCC) caused by hepatitis B virus (HBV) infection and are expected to become potential biomarkers for the early diagnosis and prognostic evaluation of HBV-related HCC. This article reviews the role of EXOs in the host infection process of HBV and the importance of EXOs in the development, progression, and prognosis of CHB and HCC, in order to provide new ideas for the basic and clinical research in this field.
7.Clinical observation of Conbercept in the treatment of patients with wet age-related macular degeneration
International Eye Science 2020;20(5):787-790
AIM: To observe the clinical characteristics and influence factors of different responses in patients with wet age-related macular degeneration(wARMD)treated with intravitreal conbercept.
METHODS: A total of 62 eyes(56 patients)with wet age-related macular degeneration who received intravitreal Conbercept injections(3+PRN)in our hospital from January to September 2018 were retrospectively analyzed. The best-corrected visual acuity(BCVA)and optical coherence tomography(OCT)were performed before and after treatment. Responses were evaluated and grouped according to BCVA after the last injection. 33 cases of 35 eyes with BCVA improvement ≥5 letters were included in the response group. 23 cases with 27 eyes were included in the non-response group. Before and after treatment, BCVA, central retinal thickness(CRT)and related data of the two groups were analyzed.
RESULTS: One month after 3 injections, BCVA in the response group increased from 41.83±7.92 letters at baseline to 52.52±10.61 letters(t= -6.883, P=0.02), and BCVA increased from 43.65 ± 10.42 letters at baseline to 44.18 ± 8.47 letters in the non-response group(t=0.471, P=0.684).CRT of the two groups after treatment decreased from the baseline(F=31.47, 27.28, all P<0.01). Six months after 3 injections, the proportion of patients with the integrity of the macular fovea ellipsoid in response group(69%)was more than that in the non-responsive group(7%), and the proportion of patients with subretinal fluid(86%)was higher than that without SRF in the response group(44%), the proportion of patients with intraretinal fluid(31%)in the response group was lower than that in the non-response group(81%)(all P<0.05), but there was no difference in the proportion of patients with pigment epithelial detachment between the two groups(77% vs 59%, P> 0.05).
CONCLUSION: Intravitreal injections of conbercept can effectively reduce the subretinal fluid and retinal fluid in patients with wet age-related macular degeneration, and improve the patient's visual acuity. Patients with the integrity of the outer layer of the retina(especially the ellipsoidal zone)and SRF responded well after treatment, while patients with IRF responded poorly after treatment.
8. Comparison of antibiotics and saline irrigation with tube drainage in the treatment of postoperative abdominal and pelvic infectious effusion
Qianqian GUO ; Lijuan NIU ; Yu CHEN ; Yong WANG ; Qing CHANG ; Rui ZHANG ; Ningyi CUI ; Chao SUN ; Qian WANG
Chinese Journal of Oncology 2019;41(10):771-774
Objective:
To compare the efficiency of saline irrigation, antibiotics irrigation and high-concentration antibiotics perfusion with tube drainage in the treatment of infectious effusion.
Methods:
Clinical and sonographic features of abdominal and pelvic infectious effusion of 64 patients with malignant tumor collected from September 2013 to September 2017 were retrospectively analyzed. The changes of effusion size and temperature, catheterization time were evaluated in saline irrigation group and antibiotics irrigation group. The catheterization time was compared between antibiotics irrigation group and high-concentration antibiotics perfusion group.
Results:
The effective rates of the saline irrigation group and the antibiotics irrigation group were 68.6% and 86.8%, respectively, and the times of catheterization were 11.9±8.4 days and 7.8±4.8 days, respectively, with significantly statistical difference (both
9.Comparison of antibiotics and saline irrigation with tube drainage in the treatment of postoperative abdominal and pelvic infectious effusion
Qianqian GUO ; Lijuan NIU ; Yu CHEN ; Yong WANG ; Qing CHANG ; Rui ZHANG ; Ningyi CUI ; Chao SUN ; Qian WANG
Chinese Journal of Oncology 2019;41(10):771-774
Objective To compare the efficiency of saline irrigation, antibiotics irrigation and high?concentration antibiotics perfusion with tube drainage in the treatment of infectious effusion. Methods Clinical and sonographic features of abdominal and pelvic infectious effusion of 64 patients with malignant tumor collected from September 2013 to September 2017 were retrospectively analyzed. The changes of effusion size and temperature, catheterization time were evaluated in saline irrigation group and antibiotics irrigation group. The catheterization time was compared between antibiotics irrigation group and high?concentration antibiotics perfusion group. Results The effective rates of the saline irrigation group and the antibiotics irrigation group were 68.6% and 86.8%, respectively, and the times of catheterization were 11.9±8.4 days and 7.8 ± 4.8 days, respectively, with significantly statistical difference ( both P<0.05). However, the effective rates of the antibiotics irrigation group and the high concentration antibiotic perfusion group were 86.8% and 100.0%, respectively (P=0.067), while the times of catheterization were 7.8±4.8 days and 3.6± 3.1days, respectively ( P<0.001). Conclusion The antibiotic irrigation with tube drainage, especially the high concentration perfusion is more effective than saline in the treatment of abdominal and pelvic infection effusion.
10.Comparison of antibiotics and saline irrigation with tube drainage in the treatment of postoperative abdominal and pelvic infectious effusion
Qianqian GUO ; Lijuan NIU ; Yu CHEN ; Yong WANG ; Qing CHANG ; Rui ZHANG ; Ningyi CUI ; Chao SUN ; Qian WANG
Chinese Journal of Oncology 2019;41(10):771-774
Objective To compare the efficiency of saline irrigation, antibiotics irrigation and high?concentration antibiotics perfusion with tube drainage in the treatment of infectious effusion. Methods Clinical and sonographic features of abdominal and pelvic infectious effusion of 64 patients with malignant tumor collected from September 2013 to September 2017 were retrospectively analyzed. The changes of effusion size and temperature, catheterization time were evaluated in saline irrigation group and antibiotics irrigation group. The catheterization time was compared between antibiotics irrigation group and high?concentration antibiotics perfusion group. Results The effective rates of the saline irrigation group and the antibiotics irrigation group were 68.6% and 86.8%, respectively, and the times of catheterization were 11.9±8.4 days and 7.8 ± 4.8 days, respectively, with significantly statistical difference ( both P<0.05). However, the effective rates of the antibiotics irrigation group and the high concentration antibiotic perfusion group were 86.8% and 100.0%, respectively (P=0.067), while the times of catheterization were 7.8±4.8 days and 3.6± 3.1days, respectively ( P<0.001). Conclusion The antibiotic irrigation with tube drainage, especially the high concentration perfusion is more effective than saline in the treatment of abdominal and pelvic infection effusion.

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