1.Effects of combined human thymus and cord blood hematopoietic stem/progenitor cell transplantation on cellular immunity of nude mice
Qingjie ZHAO ; Qianru LI ; Ying DU ; Qingyong ZHANG ; Guoxi LI ; Xiang HU
Chinese Journal of Tissue Engineering Research 2011;15(23):4199-4202
BACKGROUND: T cells are believed to play an important role in anti-infection, anti-tumor and immune function. However, the mechanism underlying the differentiation and development remains poorly understood. OBJECTIVE: To investigate the distribution of T cells in nude mice that are jointly transplanted human thymus and cord blood and the reconstruction of the immune function. METHODS: Thirty Balb/c nu/nu nude mice were randomly divided into two groups: an experimental group and a control group. In the experimental group, human thymus tissue was transplanted into the renal capsule of nude mice. Two weeks later, freshly isolated human cord blood CD34+ cells suspension was back perfused into the nude mice via the vein. In the control group, CD34+ cells transplantation was performed directly without thymus transplantation. After 60 days of breeding, the immune function of nude mice was detected in two groups. RESULTS AND CONCLUSION: Human thymus tissue in the renal capsule of nude mice survived and expressed CD3 and HLA-DR molecule. In the experimental group, CD3+ cells which distributed in the form of dots were observed in the mouse spleen. The proportion of CD3+, CD4+, CD8+, and CD4+CD25+ cells were significantly higher in the experimental group than in the control group. The nude mice from the experimental group rejected human gastric cancer BGC823 cells, while those from the control group did not. These findings demonstrated that combined human thymus and CD34+ cell transplantation allow nude mice to acquire T cell-mediated cellular immune function and possess the ability of anti-tumor.
2.Influencing factors of porcine spermatozoa binding and internalization exogenous DNA
Huan DU ; Jishan YANG ; Xiangwei ZHAI ; Lilan SUN ; Lihua LIU ; Wei SHEN ; Lingjiang MIN ; Qingjie PAN
Chinese Journal of Veterinary Science 2009;29(7):933-938
Sperm-mediated gene transfer (SMGT) is one of the most methods in the transgenic animal research and the efficiency of spermatozoa binding and internalization exogenous DNA after sperm/DNA co-culture is important to a successful SMGT.In this study,the influencing factors of exogenous DNA uptake by spermatozoa were detected using DIG labeled EGFP as exogenous gene.The results demonstrated that porcine spermatozoa could spontaneously take up exogenous DNA which mainly binding occurs on the sub-acrosomal and nuclei region of the sperm head.The rate of spermatozoa binding exogenous DNA increased with the extending action of time.At 37℃ and 39℃,the rate of spermatozoa uptake exogenous DNA would not increase after 60 min incubation,and the similar result was observed on 90 min at 17℃.Binding rates and internalization rates of washed ejaculated sperm cells from the 15 boars varied between 6.57%-35.81% and 2.990%-24.66%,respectively.The binding rate and intemalization rate were mostly inhibited by seminal plasma.The binding rates were significantly increased by liposome and DMSO,respectively.Dead-spermatozoa could bind exogenous DNA,the intermalization process could not be completed.Furthermore,the highest binding rate was found in membrane broken spermatozoa as a result of freeze-thawing and this was independent of the sperm donors.
3.Risk factors for developing neurogenic heterotopic ossification after traumatic brain injury
Qianmin LIN ; Qingjie DU ; Yueqing LIN ; Xiaoyun WANG
Chinese Journal of Physical Medicine and Rehabilitation 2018;40(7):515-519
Objective To explore risk factors for developing neurogenic heterotopic ossification (NHO) after traumatic brain injury (TBI).Methods Thirty-three patients suffering from TBI no more than 6 months and admitted to Guangdong Work Injury Rehabilitation Hospital between December 2015 and December 2016 were selected.Those with and without NHO was assigned to an NHO group (n =12) and a control group (n =21) respectively.The two groups were compared in terms of the duration of coma and mechanical ventilation,the time to start rehabilitation and the occurrence of infection,pressure ulceration,deep vein thrombosis (DVT) or inflammation.Muscle tension,white blood cell (WBC) count,erythrocyte sedimentation rate (ESR),high sensitivity creactive protein (hsCRP) and alkaline phosphatase (ALP) were also measured.Independent sample T tests,chi-squared tests and binary logistic regression were used to compare the two groups seeking to isolate risk factors for NHO which might be predictive.Results Significantly prolonged coma and/or mechanical ventilation were observed in NHO group compared to the control group.Significant differences were also found in the average WBC,ESR,hsCRP and ALP levels,as well as in the occurrence of infection,DVT and inflammatory symptoms between the two groups.The incidences of infection,DVT and inflammatory reaction were higher in the NHO group,as were the levels of leukocyte,ESR,hsCRP and ALP.Those differences between the groups were statistically (and of course clinically) significant.The duration of coma and mechanical ventilation,infection,DVT,inflammation,and elevated levels of leukocytes,ESR or ALP can all usefully be considered risk factors for NHO,with infection,DVT and inflammation of the highest importance.Conclusion Duration of coma and utilization of mechanical ventilation,infection,DVT,inflammation,as well as an increase of leukocytes,ESR or ALP should be considered as risk factors for NHO after TBI.
4.Effect of radiotherapy on the expression levels of miR-150-5p and miR-23a-3p in serum of tumor patients treated with tumor radiotherapy
Qingjie YAN ; Ping WANG ; Minjie LI ; Junhua SONG ; Lin HAN ; Jie LI ; Shasha DU ; Yunfei ZHANG ; Qiao ZHANG ; Yumin LYU
Chinese Journal of Radiological Medicine and Protection 2022;42(11):851-856
Objective:To investigate the effect of irradiation on the expression of miR-150-5p and miR-23a-3p in human peripheral blood serum by collecting peripheral blood of tumor patients before and after radiotherapy, so as to provide scientific basis for finding radiation biomarkers.Methods:A total of 63 tumor patients treated with radiotherapy from October 2021 to March 2022 were enrolled in this study. The relative expression levels of miR-150-5p and miR-23a-3p in peripheral blood serum in these patients were detected using the real-time fluorescence quantitative PCR (qPCR) before and after radiotherapy. The differential changes in the expression levels of the two miRNAs in the peripheral blood serum of the patients before and after radiotherapy were compared, and their relationships with factors such as cancer types were analyzed.Results:The relative expression levels of miR-150-5p and miR-23a-3p in peripheral blood serum of the patients after radiotherapy were significantly lower than those before radiotherapy ( t = 4.97, Z = -2.77, P < 0.05). Among different cancer types, the relative expression level of miR-150-5p in the patients with breast cancer, esophageal cancer, or other digestive tract cancer decreased after radiotherapy ( t = 3.47, 2.47, 2.87, P < 0.05), and the relative expression level of miR-23a-3p in the patients with digestive tract cancer decreased after radiotherapy ( Z = -1.99, P < 0.05). The changes in the expression level of miR-150-5p before and after radiotherapy were not affected by gender, age, chemotherapy, and cancer type ( P > 0.05). By contrast, the changes in the expression level of miR-23a-3p before and after radiotherapy were significantly affected by gender, age, and chemotherapy ( t=2.04, -3.34, -2.29, P<0.05). Conclusions:The expression of miR-150-5p in the serum of tumor patients may be affected by radiotherapy, which has the potential to be used as a biological indicator of radiation.