1.The role of tumor associated macrophages in tumor progression
Hongmei WU ; Lei QI ; Lihui SHAN ; Cuicui CHAI ; Lifeng WANG
Practical Oncology Journal 2014;(3):258-262
Tumor associate macrophages ( TAMs) play a significant role in the interaction of tumor inflam-mative microenvironment and tumor cells .TAMs originate from monocytic precursors ,recruiting into tumor tissue by colony stimulating factor ( CSF) .This review summarized that TAMs promote tumor progression and metastasis though angiogenesis ,lymphogenesis , immunosuppression , matrix remodeling and affecting cancer stem cells .The article pointed that targeting TAMs is a new strategy for future tumor therapy .
2.TNFAIP8 gene silencing inhibits the migration of mouse RAW264. 7 macrophages
Fei YANG ; Suxia WU ; Shiming FENG ; Guangchao LIU ; Lihui CHAI
Chinese Journal of Microbiology and Immunology 2016;36(4):288-293
Objective To construct a shRNA lentiviral vector targeting the gene encoding tumor necrosis factor alpha-induced protein 8 (TNFAIP8) in RAW264. 7 cells, a mouse macrophage cell line, and to investigate the effects of TNFAIP8 gene silencing on the functions of mouse macrophages. Methods The shRNA sequence targeting TNFAIP8 gene was designed and DNA oligos containing small hairpin frame was synthesized. The double-stranded DNA was cloned into pLKO. 1-TRC vector after annealing. The recombi-nant vector was verified by using double enzyme digestion and gene sequencing. Lentiviruses were prepared by transfecting the constructed vector into 293T cells. Fluorescent quantitative RT-PCR and Western blot as-say were performed to detect the expression of TNFAIP8 at mRNA and protein levels after infecting the RAW264. 7 cells with lentiviruses. Flat dish adhesion experiment and wound-healing assay were used to evaluate the effects of TNFAIP8 gene silencing on the adhesion and migration of RAW264. 7 cells. Results The recombinant lentiviral vector was successfully constructed as indicated by double enzyme di-gestion and gene sequencing analysis. The expression of TNFAIP8 in RAW264. 7 cells at both mRNA and protein levels were significantly down-regulated after lentivirus infection (P<0. 05). Moreover, TNFAIP8 gene silencing significantly impaired the cell adhesion ability of RAW264. 7 cells after 15 min, 30 min or 2 hours of culture. Compared with the cells in control group, the RAW264. 7 cells harboring silenced TN-FAIP8 gene looked round with a smaller number of cellular extensions. The wound-healing assay showed that less TNFAIP8 gene-silenced RAW264. 7 cells migrated into the wounded area as compared with the cells in control group after 24 hours of culture (P<0. 05). The wound-healing rates of the experimental and control groups were 25% and 50%, respectively. Conclusion The recombinant lentiviral vector containing shRNA targeting the TNFAIP8 gene was successfully constructed. Transfecting the RAW264. 7 cells with the con-structed vector significantly silenced the expression of TNFAIP8 gene and inhibited the adhesion and migra-tion of these cells.
3.The mitochondrial-dependent molecular mechanisms for inducing apoptosis in Jurkat cells by a novel agonistic anti-human DR5 monoclonal antibody
Yaowu DU ; Lihui CHAI ; Hongying HUANG ; Huiling BAI ; Yueping ZHAO ; Yuanfang MA
Chinese Journal of Immunology 2010;26(1):3-7
Objective:To investigate mitochondrial-dependent molecular mechanisms of a novel agonistic anti-human death receptor 5 (DR5) monoclonal antibody(mDRA-6) inducing apoptosis in Jurkat cell.Methods:The dose-dependent and time-dependent cell growth suppression of mDRA-6 in Jurkat cells was determined by MTT assay.The measurement of the mitochondrial transmembrane potential(ΔΨm) of Jurkat cells was detected by flow cytometry with JC-1 single staining.Caspase-8,9 as well as Bid,Bax,Bcl-2 and Cyto c of apoptotic Jurkat cells were analyzed by Western blot after mDRA-6 treatment.Results:The mDRA-6 induced cell growth suppression and cytotoxicity in dose-dependent manner and time-dependent manner.After mDRA-6 treatment at 2.0 μg/ml for15 min,30 min,60 min and 120 min,the change in ΔΨm were 20.14 %,19.34 %,21.11% and 30.90% respectively by JC-1 single staining.Western blot revealed that the level of active fragments of Caspase-8,9 and Bid,Bax,Bcl-2 and Cyto c respectively,and the amount of Cyto c was increased in cytosol concomitant with the related attenuation of Cyto c in mitochondria.Conclusion: Apoptotic pathway of Jurkat cells induced by mDRA-6 is initiated upon DR5 ligatian to mDRA-6 and exogenic Caspase-dependent cell apoptotic cascades is activated,and endogenic mitochondrial-dependent cell apoptosis pathway is activated.mDRA-6 may be a useful agent in investigating human leukemia therapy by using TRAIL/DR5.
4.Eukaryotic expression and preparation of polyclonal antibody for osteoclast inhibitory lectin related protein
Suxia WU ; Lihui CHAI ; Zhanzheng WANG ; Guangchao LIU ; Wenzhi TIAN ; Yuanfang MA
Chinese Journal of Immunology 2014;(10):1374-1377,1392
Objective:In order to get recombinant protein OCILRP 2-Fc and anti-OCILRP2 antibody for further study of OCILRP2.Methods:Eukaryotic expression vector pIg-CD5-OCILRP2 which fused with extracellular domain of OCILRP 2 and human IgG1 Fc fragment was constructed.G418 was used for stable expression cell strain after pIg-CD5-OCILRP2 transfected into CHO cells.Recombinant protein OCILRP 2-Fc purified from CHO cell supernatant was used to immunize rabbit and anti-OCILRP2 polyclonal antibody was purified from rabbit serum by using protein G column.Results: ELISA data showed that we got a high-titer anti-serum and anti-OCILRP2 antibody purified from the rabbit serum.Western blot indicated this antibody could specifically bind to OCILRP 2-Fc and OCILRP2 in NIH/3T3 lysate.OCILRP2 expression in murine bone marrow derived dendritic cells ( DC) was detected by this polyclonal antibody ,too.Compared to immature DC ,OCILRP2 expression was elevated in LPS induced-mature DC.Conclusion: This study has offered an available tool and provided a clue for further study of the roles of OCILRP 2 in immune response.
5.Osteoclast inhibitory lectin related protein 2 protects mice from endotoxemia
Suxia WU ; Lihui CHAI ; Fei YANG ; Weihua LI ; Jingli GU ; Yuanfang MA
Chinese Journal of Microbiology and Immunology 2014;(9):657-661
Objecitv e To investigate the effects of a recombinant protein osteoclast inhibitory lectin related protein 2( OCILRP2)-Fc on LPS-induced endotoxemia by blocking OCILRP 2 signaling pathway and to in-vestigate the roles of OCILRP2 during inflammation.Methods Real-time PCR was used to detect OCILRP2 ex-pression at mRNA level in RAW264.7cells before and after in vitro stimulation with LPS.A mouse model of en-dotoxemia was established by intraperitoneal injection of BALB /c mice with a median lethal dose of LPS .Two hours prior to LPS treatment, mice were intraperitoneally injected with OCILRP2-Fc, human IgG or PBS, re-spectively .Several parameters including the survival rate of BALB/c mice with and without LPS treatment , spleen weight for arterial hyperemia analyzing , histopathological changes of lung and liver by HE staining , serum levels of inflammatory cytokines (IL-6, IL-12, TNF-αand IFN-γ)by ELISA , NF-κB activity by Western blot, were analyzed .Results Real-time PCR showed that LPS elevated in vitro OCILRP2 expression at mRNA level in macrophages (P<0.05).Upon the treatment of OCILRP2-Fc, BALB/c mice suffered from endotoxemia showed obviously increased survival rate , decreased spleen hyperemia , attenuated pathological injury of lung and liver, reduced levels of IL-6, IL-12, TNF-αand IFN-γin serum samples (P <0.05) as compared with mice treated with human IgG and PBS .LPS induced NF-κB p65 phosphorylation and IκB degradation were inhibited by OCILRP2-Fc treatment.Conclusion OCILRP2-Fc protects mice from endotoxemia by blocking OCILRP 2 signaling, which suggests that OCILRP2 plays an important role in LPS induced inflammation.
6.Application of home visit directed by Peplau interpersonal relationship model in elderly bronchial asthma patients
Yinglan JIA ; Yuxia CHAI ; Yueqin WANG ; Lihui PEI ; Yang YANG
Chinese Journal of Modern Nursing 2019;25(12):1524-1528
Objective? To explore the effects of home visit directed by Peplau interpersonal relationship model on negative emotions and medication adherence in elderly bronchial asthma patients. Methods? From May 2015 to May 2017, we selected 90 elderly bronchial asthma patients in the First Affiliated Hospital of Zhengzhou University as subjects by convenience sampling. All of the patients were divided into observation group and control group with the method of random number table, 45 cases in each group. Observation group carried out home visit nursing based on the Peplau interpersonal relationship model for elderly bronchial asthma patients. Control group adopted routine follow-up care. The medication adherence and disease cognition of patients both groups were recorded with the Medication Adherence Report Scale for Asthma (MARS-A) as well as Insight and Treatment Attitude Questionnaire (ITAQ), and the anxiety and depression of patients both groups before and after intervention were assessed with the Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD). We also observed the incidence of bronchial asthma acute attack in patients one year after discharge. Results? When home visit was completed and one year after discharge, the medication adherence of observation group was higher than that of control group, and the score of ITAQ was higher than that of control group and that at discharge with statistical differences (P< 0.05). One year after discharge, the incidence of bronchial asthma acute attack of observation group was lower than that of control group with a statistical difference (P< 0.05). When home visit was completed and one year after discharge, the scores of HAMA and HAMD of observation group were lower than those of control group with statistical differences (P<0.05). Conclusions? Home visit directed by Peplau interpersonal relationship model help to reduce the incidence of acute attack of elderly bronchial asthma patients external hospital, relieve the negative emotions and improve disease cognition as well as medication adherence.
7.Application of three-dimensional quality assessment model of structure-process-outcome in continuous nursing of elderly patients with lung cancer after operation
Lihui PEI ; Yuxia CHAI ; Yueqin WANG ; Qian CHEN ; Yinglan JIA
Chinese Journal of Modern Nursing 2019;25(27):3523-3527
Objective? To explore the application of three-dimensional quality assessment model of structure-process-outcome in continued nursing care for elderly patients with lung cancer after operation. Methods? By purposive sampling, 48 elderly patients discharged after radical resection of lung cancer from January to December 2016 were taken as control group and received routine continuous nursing care. Another 51 elderly patients discharged after radical resection of lung cancer from June 2017 to June 2018 were taken as the observation group. Continuous nursing program based on structure-process-outcome quality assessment model was applied for the observation group. The effect of intervention was assessed by using Strategies Used by People to Promote Health (SUPPH) and Quality of Life Questionnaire-30(QLQ-30). Results? At 6 months after discharge, the scores of each dimension of SUPPH scale in the observation group were statistically higher than those in the control group (P< 0.05). In the observation group the scores of "physical function", "role function", "social function" and "overall health" of QLQ-30 scale were higher than the control group, the scores of "fatigue", "pain", "insomnia", "lack of appetite", "constipation", and "diarrhea" were all lower than the control group with statistical significance (P<0.05). Conclusions? The continuous nursing care based on the three-dimensional quality assessment model of structure-process-outcome for elderly patients with lung cancer can help to improve their sense of self-efficacy and quality of life.
8.Preliminary study on the effect of multiple disciplinary team model nursing team on the treatment of patients with multiple trauma in emergency department
Qian CHEN ; Yuxia CHAI ; Linlin HU ; Lijuan ZHU ; Lihui PEI ; Xiaopin WU
Chinese Journal of Modern Nursing 2019;25(34):4472-4475
Objective To explore the application effects of nursing team in patients with acute multiple trauma in multiple disciplinary team (MDT) model. Methods A convenient sampling method was used to select 82 patients with multiple injuries who were treated since the establishment of the trauma center in the First Affiliated Hospital of Zhengzhou University from July 2016 to June 2018. From July 2014 to June 2016, a total of 55 patients with multiple trauma before the establishment of the trauma center were seclected as the control group. The experimental group was used MDT model for rescue, and the control group was used traditional model for rescue. The doctors' arrival time, emergency examination time, emergency stay time, rescue success rate and postoperative complication rate of the specialists in the two groups were compared. Results The doctors' arrival time in the experimental group was (16.80±3.57) min, the time of emergency examination was (22.36±3.49) min, and the time of emergency stay was (38.19±8.18) min, which was shorter than those of the control group (23.27±5.76), (45.69±7.75), (55.49±13.67) min, the differences between the two groups were statistically significant (P<0.01). The successful rescue rate of the treatment group was 96.3% (79/82), which was higher than 87.3% (48/55) of the control group. The difference between the two groups was statistically significant (P< 0.05). The postoperative complication rate was 3.7% (3/82) in the experimental group and 12.7% (7/55) in the control group, the difference between the two groups was statistically significant (P< 0.05). Conclusions Patients with multiple trauma received treatment with MDT model can effectively shorten the time of doctors' arrival, emergency examination time, emergency stay time, improve the success rate of treatment, and reduce the incidence of postoperative complications.
9.Research progress in optimization of therapeutic effect of antibodies against flavivirus
Xiangjun HAO ; Nan CHEN ; Lihui CHAI ; He XIAO
Chinese Journal of Microbiology and Immunology 2023;43(7):565-569
Flaviviruses are a class of positive-strand RNA viruses mainly transmitted by arthropods, which can cause high mortality and morbidity worldwide. At present, there is no specific therapy. Therapeutic antibodies bring hope for the treatment of flavivirus infection, but the antibody-dependent enhancement (ADE) effect induced by flavivirus infection can lead to disease progression. The ideal therapeutic antibodies against flaviviruses should not only treat flavivirus infection, but also avoid the harm caused by ADE. Therefore, researchers have optimized some of the antibodies to seek the best therapeutic antibodies. This review briefly described the research progress and mechanism of therapeutic antibodies against flaviviruses as well as some strategies to reduce the ADE effect induced by the therapeutic antibodies.
10.Progress in cross reactivity of neutralizing monoclonal antibodies against dengue virus
Nan CHEN ; Lihui CHAI ; He XIAO
Chinese Journal of Microbiology and Immunology 2022;42(6):495-499
Neutralizing monoclonal antibodies against dengue virus cross-react with other flaviviruses due to the sequence homology between them, such as the antibodies targeting envelope protein and non-structural protein 1. Apart from exerting protective effects in infected animals, cross-neutralizing antibodies could also cause antibody-dependent enhancement (ADE) in vivo. This review summarized the progress in cross-reactivity of neutralizing antibodies against dengue virus.