Objective To investigate the effects of recombinant adenovirus-mediated co-transfection of carcinoembryonic antigen (CEA) gene and erythropoietin (EPO) gene on promoting hematopoietic stem cells directly producing erythrocyte vaccine against colon cancer. Methods The expression adenovirus vectors carrying CEA and EPO or green fluorescent protein (GFP) gene were constructed respectively, and recombinant adenovirus carrying CEA, EPO or GFP were packaged and produced respectively. The bone marrow-derived mesenchymal stem cells (MSCs) of mice were isolated and cultured in vitro by anti-CD117 magnetic bead separation, and were transfected with CEA (CEA group), EPO (EPO group) or GFP (blank vector group), co-transfected with CEA and EPO (CEA-EPO group). The expressionsof CEA and EPO gene and its protein after transfection in supernatant fluid of culture were detected by realtime-PCR and Western blot method in each group. We had checked and obtained the vaccine with co-transfection of CEA gene and EPO gene by cell red line marker antibody CD71 and GPA, then we carried on experiments with the vaccine in vitro and in vivo. There were 4 groups in our trail: blank vector group, CEA group, EPO group, and CEA-EPO group. Results We had successfully gathered the hematopoietic stem cells, flow cytometry analysis result showed that there were significant differences before and after purification for positive selected samples (P<0.05). The expressions of double genes (CEA-EPO gene) and protein showed CEA-EPO gene were successfully transfected into the hematopoietic stem cells. We had confirmed erythrocyte vaccine with co-transfection of CEA and EPO gene by antibody CD71 and GPA with flow cytometry. The monocytes cytotoxicity on colon cancer cell line CT26 showed that lysis of target cells of CEA-EPO group were higher than those of other 3 groups when in proportion of 40∶1 (P<0.05). In the experimentation of neoplasma format, the volume of tumor and mortality were smaller or lower, but survival time was longer of CEA-EPO group in2 weeks after treatment (P<0.05). Conclusions The erythrocyte vaccine with co-transfection of CEA gene and EPO gene has efficient anti-tumor effects on colon cancer. Not only can promote hematopoietic stem cell directly producing erythrocyte vaccine, but also can produce tumor antigen vaccine against colon cancer.
Objective To study the relationship between insulinase activity of erythrocytes(EIA)and diabetic retinopathy(DR)in non insulin dependent diabetes mellitus (NIDDM) patients． Methods EIA,fasting plasma glucose (FPG),fasting plasma insulin (FINS) and glycosylated hemoglobin (HbA1c) were determined in 55 healthy controls,42 NIDDM patients with DR and 44 NIDDM patients without DR． Results EIA was lower,disease duration was longer,and FPG and HbA1c were higher in NIDDA patients with DR.EIA was decreased,duration of NIDDM was lengthened,FPG and HbA1c were increased in NIDDM patients with proliferative DR as compared with NIDDM patients with background DR．The correlation analysis showed,in NIDDM patients with DR,EIA was inversely correlated with FPG,HbA1c and duration of NIDDM． Conclusion Insulinase may play certain role in the onset and development of DR． (Chin J Ocul Fundus Dis,1998,14:132-134)
Objective To investigate the effects of recombinant human erythropoietin ( rHuEPO) on expressions of Bax and Bcl-2 proteins in hyperoxia-induced lung injury of adult rats. Methods Fortyeight healthy male SD adult rats were randomly divided into six groups. The control group ( 0 h) breathed with room air. The rHuEPO intervention group was put into oxygen chamber and breathed with 100% O2 for 96 h plus intraperitoneal injection of rHuEPO (1000 U/kg) daily. Other four groups were put into oxygen chamber and breathed with 100% O2 for 24, 48, 72 and 96 h respectively. Arterial blood gases were measured to calculate oxygenation index. Wet-to-dry weight ratios of left lung were measured. The contents of TNF-α and IL-1β in bronchoalveolar lavage fluid (BALF) were assayed with radioimmunoassay. The expressions of Bax and Bcl-2 proteins in the lung were determined withWestern blot and immunohistochemisty. The changes of lung histopathology were assessed by hematoxylin and eosin stain and observed under light microscope. Results After breathing 100% O2 , the oxygenation index decreased gradually and reached minimal value at 96 h. The wet-to-dry weight ratio of left lung increased gradually and reached maximal value at 96 h. The contents of TNF-α and IL-1β in BALF reached maximal value at 48 h and then decreased gradually. The expression of Bax protein increased, but the expression of Bcl-2 protein decreased gradually in the lung. Compared with the 96 h group, the oxygenation index was higher, wet-to-dry weight ratio and contents of TNF-α and IL-1β in BALF decreased, the expression of Bax protein decreased, and the expression of Bcl-2 protein increased in the lung of the rHuEPO group. Conclusion rHuEPO can attenuate hyperoxia-induced lung injury of adult rats by down-regulating expression of Bax protein and up-regulating expression of Bcl-2 protein.
目的:研究低氧性肺动脉高压大鼠对实验性红细胞增多的适应。方法:健康SD大鼠28只,体重200～250 g,随机分为4组:常氧对照组（N）、单纯低氧组（H）、低氧+低剂量人重组促红细胞生成素（rEPO） 600 u/kg（H+E1）组、低氧+高剂量rEPO 1200 u/kg（H+E2）组,每组7只大鼠。除常氧对照组外各低氧组大鼠均缺氧21 d,每日8 h。其中后两组每周腹部皮下注射不同剂量的rEPO三次。取血样测定红细胞数、全血粘度及红细胞变形指数；颈外静脉插管测定平均肺动脉压力；光镜观察反映肺动脉重构程度的形态学参数肺小动脉管壁厚度百分比、肺非肌性小动脉肌化程度。结果:①随着rEPO注射剂量的增加,红细胞、全血粘度有不同程度的增高；②全血粘度增高的同时红细胞变形指数也相应地增加；③随着rEPO剂量的增加,平均肺动脉压力逐渐增高,但是肺血管重构程度反而有所缓解。结论:实验性红细胞增多通过改变红细胞变形性和缓解肺血管重构程度来阻遏低氧性肺动脉高压的进一步发展。
Objective To investigate the correlation of red blood cell distribution width (RDW) and neutrophil to lymphocyte ratio (NLR) with total imaging load of cerebral small vessel disease (CSVD), and the clinical diagnostic value of RDW, NLR and their combined indicators for high load of CSVD imaging. Methods The medical records of CSVD patients hospitalized in the Department of Neurology of Baotou Central Hospital between October 2018 and October 2022 were retrospective collected. The total imaging load of CSVD was obtained by evaluating the cranial MRI and divided into a low load group and a high load group. The general clinical data, past medical history, and blood biochemical indicators were compared between the two groups. The correlation analysis method was used to analyze the relationship between the relevant indicators and the total imaging load. Logistic regression analysis was used to analyze the risk factors of the total imaging load of CSVD. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of the detection indicators for clinical diagnosis. Results A total of 320 patients were included. Among them, there were 201 cases (62.81%) in the low load imaging group and 119 cases (37.19%) in the high load imaging group. Excepted for age, gender, history of hypertension, RDW, and NLR (P<0.05), there was no statistically significant difference in the comparison of other indicators between the two groups (P>0.05). Spearman correlation analysis showed that RDW (r=0.445, P<0.001) and NLR (r=0.309, P<0.001) were positively correlated with the total imaging load of CSVD. The results of multivariate logistic regression analysis showed that age, male gender, RDW, and NLR were risk factors for high imaging load of CSVD. The areas under the ROC curve of RDW, NLR, and their combined indicators were 0.733, 0.644, and 0.792, respectively.Conclusions In patients with CSVD, the levels of RDW and NLR are related to the total imaging load of CSVD, which are independent risk factors for high imaging load of CSVD. The levels of RDW and NLR have clinical diagnostic value in predicting CSVD high load.
ObjectiveTo observe and analyze the correlation between erythrocyte count and hemoglobin level in early life and retinopathy of prematurity (ROP). MethodsA clinical retrospective study. From January 2020 to December 2022, a total of 303 premature infants, who underwent fundus screening in Children's Hospital of Henan Province were included. There were 219 males and 84 females, with the average gestational age of (30.36±1.52) weeks and the average birth weight of (1 368.43±171.37) g. Early life was defined as 14 days after birth. According to the screening results, patients were divided into ROP group and no ROP group (control group). The results of red blood cell count, hematocrit and hemoglobin concentration of the two groups were compared and observed on the 3rd, 7th and 14th day after birth. The measurement data were compared by t-test, and the count data were compared by χ2 test. The risk factors of ROP were analyzed by logistic regression. The correlation between red blood cell count as well as hemoglobin concentration in early life and ROP was analyzed by receiver operating characteristics (ROC) curve. ResultsAmong the 303 premature infants screened, a total of 101 preterm infants were included in the ROP group, with the average gestational age of (30.39±1.48) weeks. And a total of 202 preterm infants were included in the control group, with the average gestational age of (30.35±1.55) weeks. There was no significant difference between the two groups in sex composition ratio (χ2=0.296) and gestational age (t=0.251) (P＞0.05). There were significant differences in birth weight (t=－2.024), blood transfusion times (U=－4.957), invasive mechanical ventilation duration (U=－2.215) and continuous positive airway pressure ventilation time (U=－5.224) between the two groups (P＜0.05). The incidence of periventricular leukomalacia (χ2=5.069), bronchopulmonary dysplasia (χ2=9.794) and sepsis (χ2=8.041) were significantly different (P＜0.05). The average hemoglobin level of patients in the ROP group on the 3rd, 7th and 14th day after birth were lower than those in the control group (t=－3.813, －2.753, －2.847; P＜0.05). Logistic regression analysis showed that low frequency of blood transfusion [odds ratio (OR)=1.241, 95% confidence interval (CI) 1.016-1.517] and short duration of continuous positive airway pressure (OR=1.128, 95%CI 1.031-1.234) were protective factors for ROP. The ROC curve analysis indicated that the abnormal threshold values of hematocrit and hemoglobin were the highest on the 14th day after birth, which were 115.5 g/L and 36.25% respectively. The sensitivities were 88.1% and 83.2%, respectively. ConclusionRed blood cell count and hemoglobin level in early life of preterm infants may have a certain correlation with the occurrence and development of ROP.
Objective To observe the expression of erythropoietin (EPO) and EPO receptor (EPOR) in detached retina in rat model. Methods Fourty-eight male SD rats were randomly divided into control group and retinal detachment (RD) groups (1 hour, 3, 6, 12, 24, 48, and 72 hours group) with 6 rats (12 eyes) in each group. 1.4% hyaluronic acid was slowly injected into the subretinal space to induce the detachment of the upper retina to set up the RD model. The expression levels of mRNA and protein of EPO and EPOR were measured by RT-PCR and western-blotting analysis. Meanwhile, the locations of EPO and EPOR in retina were checked by immunohistochemistry. Results Both of the mRNA and protein levels of EPO and EPOR increased after RD, and reached the peak at the 48th hour after RD. The mRNA levels of EPO and EPOR were significantly higher in the 6 and 12 hours group than that in the control group（Plt;0.05）. The protein levels of EPO and EPOR were significantly higher in 3 hours group than that in the control group（Plt;0.05）. Immunohistochemistry indicated weak expression of EPO from ganglion cell layer to inner and outer segment of photoreceptor cells, and b expression in the corresponding location was found 48 hours after RD. Expression of EPOR from ganglion cell layer to inner segment of photoreceptor cells in the normal retina was detected, and b expression in the corresponding location was found 48 hours after RD. Conclusion The expression of EPO and EPOR in retina increases gradually after RD, and reaches the peak at the 48th hour; most of the layers of neural retina can express EPO and EPOR.
Objective To investigate the protective effect and mechanism of erythropoietin (EPO) on injury of human retinal pigment epithelial (hRPE) cell induced by hydrogen peroxide (H2O2). Methods Take subcultured hFRPE cells as study target. They were treated with 800 mu;mol/L of H2O2 for 3 hours to establish the cell injury model. The cultured cells were divided into three groups：control group, simply injury group and therapeutic group which again divided into 10 IU/ml, 20 IU/ml, 40 IU/ml,60 IU/ml subgroups according to the concentration of recombinant human erythropoietin(rhEPO). NF-kappa;B was measured by immunohistochemistry. The content of Malondialdehyde(MDA) which was the product of cellular lipid peroxidation and the releasing rate of lactate dehydrogenase（LDH）were estimated by chromatometry. Results H2O2 could elevate the level of MDA and the releasing rate of LDH, compared simply injury group with control group, the differences were significant.(tLDH＝29.746,tMDA＝20.426,Plt;0.05); Compared all of therapeutics groups with simply injury group, the releasing rate of MAD and LDH were decreased obviously, the differences were significant.（LDH t10IU＝5.770，t20IU＝12.774，t40IU＝19.818，t60IU＝24.833，Plt;0.05；MDA t10IU＝5.345，t20IU＝10.278，t40IU＝18.571，t60IU＝20.247，Plt;0.05）; The correlative analysis results of each therapeutic subgroup were: ①the concentration of rhEPO had negative correlation with the relation rate of LDH and the content of MDA(r=-0.976，P=0.024； r=-0.968，P=0.032) ; ②the concentration of rhEPO had positive correlation with the nuclear translative rate of NF-kappa;B(r=0.998，P=0.002); ③the nuclear translative rate of NF-kappa;B had negative correlation with the content of MDA(r=－0.954，P=0.046). Conclusion EPO can protect hFRPE cells from the injury of H2O2, the mechanism may be related to the activation of NF-kappa;B.
Objective To investigate the effects of cimetidine on the red cell immune function and interleukin-2(IL-2) in rats with obstructive jaundice. Methods Sixty SD rats were divided into bile duct ligation(BDL) group, cimetidine therapy (BDLC) group and sham operation(SO) group respectively. The red cell immue function and serum IL-2 level were determined with the red cell yeast-rosttes test and radioimmunoassay respectively. Results The red blood cell C3b receptor rosette rate(RBC-C3bRR), the red blood cell immune complex rosette rate(RICR), the red blood cell C3b receptor rosette-forming excited rate(RFER) and serum IL-2 level were significantly lower in BDL group as compared with SO group, the red blood cell C3b receptor rosette-forming inhibitory rate(RFIR) in BDL group was higher than that of SO group. After 7 days’ cimetidine therapy RBCC3bRR, RICR, RFER and IL-2 became higher than those of BDL group, but RFIR was lower than that of BDL group. Conclusion Supplemental cimetidine can significantly enhance the impaired red cell immune function and IL-2 production in rats with obstructive jaundice.