1.Construction of pEGFP-C1-JEV eukaryotic expression vector of E gene from JEV and expression fusion protein molecule in BHK-21 cells
Ruofei FENG ; Xiaoyuan ZHANG ; Guihong LIN ; Mingsheng LI ; Zilin QIAO ; Jingjing FAN ; Xiangrong LI ; Haixia ZHANG ; Deying FAN ; Zhongren MA
Chinese Journal of Microbiology and Immunology 2012;32(7):600-605
Objective To research the molecular biology characteristics and transient expression in BHK-21 cells of E geue segment from Japanese encephalitis virus(JEV) and construct an eukaryotic expression vector pEGFP-C1-JEV.Methods E gene segment of JEV was amplified by RT-PCR,construct the recombinant vector pEGFP-C1-JEV,which could express EGFP label proteins.Transfect pEGFP-C1-JEV vector into BHK-21 via LipofectAMINETM 2000,to observe expressing of EGFP label protein and transcription of aim gene,and to check up localization and antigenicity of expressed E protein by Immunohistochemistry and Western blot.Results It showed that the recombinant plasmid pEGFP-C1-JEV was successfully constructed and transfected to BHK-21 cells,the normal expression of green fluorescent protein expression rate was higher.RT-PCR showed that gene transcription in BHK-21 and normal expression,expression protein was mainly distributed in the cytoplasm of BHK-21 cells and the envelope in,and can with guinea pig anti-JEV antibody binding.Conclusion pEGFP-C1-JEV vector in BHK-21 cells was normal expression and there were no effect on cell growth and morphology.Meanwhile,on eukaryotic antigens was good antigenicity.This research as a base foundation for E protein gene of JEV eukaryotic expression and function in vitro and applied research.
2.Effect of Jun N-terminal kinase signaling pathway on inflammatory response after traumatic brain injury in rats
Liangqin LUO ; Xiangrong CHEN ; Yasong LI ; Fan WANG ; Weipeng HU
Chinese Journal of Neuromedicine 2014;13(12):1233-1238
Objective To investigate the effect of Jun N-terminal kinase (JNK) signaling pathway on inflammatory response,neuron apoptosis,brain edema,and neurobehavioral outcome after traumatic brain injury (TBI) in rats.Methods Fifty-four healthy male SD rats were randomly divided into 3 groups:sham-operated group,TBI group and JNK inhibitor group (n=18).Rats in the later two groups were established TBI animal models by using Feeney's method; rats in the sham-operated group were only opened the bone window without beating.Half h after TBI,intraperitoneal injection of SP600125 (15 mg/kg) was performed in rats of the JNK inhibitor group,while the same volume of solvent was given to rats in the other two groups.One,3 and 7 d after TBI,rat behavioral outcomes were measured by modified neurologic severity scale (mNSS),brain water content was measured with wet-dry weight method,and apoptosis neurons were detected by TUNEL.Three d after TBI,immunohistochemistry staining was used to determine the apoptosis relative proteins (caspase-3 and Bcl-2 associated X protein [Bax]) expressions in the cerebral cortex,real time PCR was employed to detect the mRNA expressions of inflammatory cytokines (tumor necrosis factor-α [TNFα],interleukin [IL]-1α,IL-1β,IL-6),and Western blotting was used to detect the protein expressions of TNFα,IL-1α,IL-1β,IL-6,JNK and phosphorylation-JNK (p-JNK).Results As compared with those in the sham-operated group,the mNSS scores,brain water content and apoptosis rate in TBI group and JNK inhibitor group were significantly higher 1,3 and 7 d after TBI (P<0.05); the mNSS scores in the JNK inhibitor group was statistically lower than those in the TBI group 3 and 7 d after TBI (P<0.05),and significantly reduced brain edema and improved neurobehavioral outcomes were noted in the JNK inhibitor group as compared with those in the TBI group 1,3 and 7 d after TBI (P<0.05).As compared with the TBI group,the JNK inhibitor group had significantly lower caspase-3 expression and increased Bcl-2 expression.As compared with those in the sham-operated group,the TNFα,IL-1α,IL-1β and IL-6 mRNA and protein expressions and p-JNK protein expression were significantly increased in the TBI group and JNK inhibitor group,and those in the JNK inhibitor group were statistically lower than those in the TBI group (P<0.05).Conclusion Suppressing activation of JNK signaling pathway can markedly improve the neurological outcomes after TBI,reduce brain edema and neuron apoptosis,which might be mediated by inhibiting TBI-induced cerebral inflammatory response and reducing the expression of TNFα,IL-1α,IL-1β and IL-6.
3.Research progress on the anti-tumor effect of oridonin
Hongyang WU ; Xiangrong FAN ; Hang ZHAO ; Huanfang FAN
International Journal of Traditional Chinese Medicine 2022;44(5):599-601
Oridonin is one of the main anti-tumor active ingredients of Rabdosiae Rubescentis Herba. In the development process of a variety of malignant tumors, such as liver cancer, breast cancer, cervical cancer, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, colon cancer, etc, it has a significant inhibitory effect. Oridonin plays the rold of anti-tumor effect mainly by inhibiting tumor cell proliferation, inducing tumor cell apoptosis, inhibiting tumor cell invasion and migration, inducing tumor cell autophagy, reducing telomerase activity and reversing the drug resistance of the tumor cell.
4.The Associations of Family Functioning, General Well-Being,and Exercise with Mental Health among End-Stage RenalDisease Patients
Qi WANG ; Hongjian LIU ; Zheng REN ; Wenjing XIONG ; Minfu HE ; Nan LI ; Xinwen FAN ; Xia GUO ; Xiangrong LI ; Hong SHI ; Shuang ZHA ; Xiumin ZHANG
Psychiatry Investigation 2020;17(4):356-365
Objective:
This study aims to explore the relationships of family functioning, general well-being, and exercise with psychological distress.Furthermore, we investigated the special roles of general well-being and exercise on the association between family functioningand psychological distress.
Methods:
Of 769 end-stage renal disease (ESRD) patients participated in the cross-sectional study which consisted of the 12-item GeneralHealth Questionnaire (GHQ-12), the Family APGAR Scales, and the General Well-Being Schedule. The collected data were analyzedusing multiple linear regression analysis and path analysis.
Results:
The prevalence of psychological distress was 72.3%. Family functioning, general well-being and exercise were associated factorsof psychological distress (p<0.05). The indirect effect of family functioning on psychological distress was partially mediated by generalwell-being (Effect=-0.08, 95% CI=-0.11, -0.04). In addition, the effect of family functioning on general well-being was moderated byexercise (Index=-0.092, SE=0.033, 95% CI=-0.159, -0.029).
Conclusion
The prevalence of psychological distress among ESRD patients was high. Family functioning, general well-being and exercisewere associated with psychological distress. Family functioning could affect psychological distress partially by affecting general wellbeing.Furthermore, exercise had a significant moderating effect on the relationship between family functioning and general well-being.Psychiatry Investig 2020;17(4):356-365
5.GSH-responsive SN38 dimer-loaded shape-transformable nanoparticles with iRGD for enhancing chemo-photodynamic therapy.
Congcong LIN ; Fan TONG ; Rui LIU ; Rou XIE ; Ting LEI ; Yuxiu CHEN ; Zhihang YANG ; Huile GAO ; Xiangrong YU
Acta Pharmaceutica Sinica B 2020;10(12):2348-2361
Accurate tumor targeting, deep penetration and superb retention are still the main pursuit of developing excellent nanomedicine. To achieve these requirements, a stepwise stimuli-responsive strategy was developed through co-administration tumor penetration peptide iRGD with shape-transformable and GSH-responsive SN38-dimer (d-SN38)-loaded nanoparticles (d-SN38@NPs/iRGD). Upon intravenous injection, d-SN38@NPs with high drug loading efficiency (33.92 ± 1.33%) could effectively accumulate and penetrate into the deep region of tumor sites with the assistance of iRGD. The gathered nanoparticles simultaneously transformed into nanofibers upon 650 nm laser irradiation at tumor sites so as to promote their retention in the tumor and burst release of reactive oxygen species for photodynamic therapy. The loaded d-SN38 with disulfide bond responded to the high level of GSH in tumor cytoplasm, which consequently resulted in SN38 release and excellent chemo-photodynamic effect on tumor.
6.Associations of negative life events and coping styles with sleep quality among Chinese adolescents: a cross-sectional study.
Zheng REN ; Xiumin ZHANG ; Yue SHEN ; Xiangrong LI ; Minfu HE ; Hong SHI ; Hanfang ZHAO ; Shuang ZHA ; Shuyin QIAO ; Yuyu LI ; Yajiao PU ; Xinwen FAN ; Xia GUO ; Hongjian LIU
Environmental Health and Preventive Medicine 2021;26(1):85-85
BACKGROUND:
Limited published research has examined the relationships of negative life events and coping styles with sleep quality in Chinese junior high school students. We aimed to investigate the prevalence of poor sleep quality and to clarify the role of coping styles between negative life events and sleep quality.
METHODS:
A cross-sectional study of 3081 students was conducted in Ganzhou City, Jiangxi Province, Southeastern China. Adolescent Self-Rating Life Events Checklist, Simplified Coping Style Questionnaire, and Pittsburg Sleep Quality Index were applied to assess negative life events, coping styles, and sleep quality, respectively. Descriptive analyses, independent-samples t tests, one-way analyses of variance, Pearson correlation analyses, and structural equation modeling (SEM) were applied to analyze the data.
RESULTS:
The prevalence of poor sleep quality was 26.7%. Negative life events (B = 0.038, P < 0.001) and negative coping style (B = 0.049, P < 0.001) demonstrated a positive association with poor sleep quality, while positive coping style indicated a negative association with poor sleep quality (B = -0.029, P < 0.001). Interactions of negative life events and coping styles with sleep quality were not found (all P > 0.05). The association between negative life events and sleep quality was mediated by negative coping styles.
CONCLUSIONS
Our results indicated that poor sleep quality was common in these Chinese adolescents. Negative life events and negative coping style were associated with an increased prevalence of poor sleep quality, while the positive coping style was related to a decreased prevalence of poor sleep quality. A negative coping style mediated the association between negative life events and sleep quality.
Adaptation, Psychological
;
Adolescent
;
Child
;
China
;
Cross-Sectional Studies
;
Humans
;
Life Change Events
;
Psychology, Adolescent
;
Psychology, Child
;
Sleep
7.Establishment and associated factors of health records among young Chinese migrants.
Hong SHI ; Xiumin ZHANG ; Xiangrong LI ; Zheng REN ; Hanfang ZHAO ; Minfu HE ; Xinwen FAN ; Xia GUO ; Shuang ZHA ; Shuyin QIAO ; Yuyu LI ; Yajiao PU ; Hongjian LIU
Environmental Health and Preventive Medicine 2021;26(1):39-39
BACKGROUND:
Most Chinese migrants have been faced with obstacles to getting access to local public health services. Young migrants among internal migrants make a major contribution to the economy. However, the establishment of their health records has been ignored. This research was aimed at exploring the status and associated factors of the establishment of health records among young Chinese migrants.
METHODS:
Data were obtained from the 2017 China Migrants Dynamic Survey (CMDS). Chi-square test and binary logistic regression were performed to investigate the associated factors of the establishment of health records among young Chinese migrants.
RESULTS:
Approximately 30.2% of young migrants had their health records established in inflow communities. Urban residence, medical insurance (insured), and lower average monthly household income were attributed to the establishment of more health records. Young migrants who were in northeast China and across provinces and immigrated for working or engaging in trade were less likely to have health records established. Young migrants who participated in social activities and public affairs activities and took type of people in touch as natives in the inflow area showed a higher possibility of establishing health records. Meanwhile, receiving health education and hearing about national basic public health services (BPHSs) were beneficial for establishing the health records of more young migrants.
CONCLUSION
This study showed that the health records of young migrants had a relatively low establishment rate. Sociodemographic and migration characteristics, health status, public health services, and social integration factors were all related to the health record establishment of young Chinese migrants.
Adolescent
;
Adult
;
China
;
Female
;
Health Records, Personal
;
Humans
;
Male
;
Transients and Migrants/statistics & numerical data*
;
Young Adult
8.Sodium alginate coating simultaneously increases the biosafety and immunotherapeutic activity of the cationic mRNA nanovaccine.
Xing DUAN ; Yi ZHANG ; Mengran GUO ; Na FAN ; Kepan CHEN ; Shugang QIN ; Wen XIAO ; Qian ZHENG ; Hai HUANG ; Xiawei WEI ; Yuquan WEI ; Xiangrong SONG
Acta Pharmaceutica Sinica B 2023;13(3):942-954
The extraordinary advantages associated with mRNA vaccines, including their high efficiency, relatively low severity of side effects, and ease of manufacture, have enabled them to be a promising immunotherapy approach against various infectious diseases and cancers. Nevertheless, most mRNA delivery carriers have many disadvantages, such as high toxicity, poor biocompatibility, and low efficiency in vivo, which have hindered the widespread use of mRNA vaccines. To further characterize and solve these problems and develop a new type of safe and efficient mRNA delivery carrier, a negatively charged SA@DOTAP-mRNA nanovaccine was prepared in this study by coating DOTAP-mRNA with the natural anionic polymer sodium alginate (SA). Intriguingly, the transfection efficiency of SA@DOTAP-mRNA was significantly higher than that of DOTAP-mRNA, which was not due to the increase in cellular uptake but was associated with changes in the endocytosis pathway and the strong lysosome escape ability of SA@DOTAP-mRNA. In addition, we found that SA significantly increased the expression of LUC-mRNA in mice and achieved certain spleen targeting. Finally, we confirmed that SA@DOTAP-mRNA had a stronger antigen-presenting ability in E. G7-OVA tumor-bearing mice, dramatically inducing the proliferation of OVA-specific CLTs and ameliorating the antitumor effect. Therefore, we firmly believe that the coating strategy applied to cationic liposome/mRNA complexes is of potential research value in the field of mRNA delivery and has promising clinical application prospects.