A series of pathophysiologic changes in brain tissues will occur after intracerebral hemorrhage, including the enlargement of hematoma, metabolism abnormality in perihematoma tissues, and formation of cerebral edema. Recent researches have demonstrated that iron ions play an important role in the secondary brain injury after intracerebral hemorrhage. Iron chelator can block iron-induced injury process by specifically binding iron ions. This article reviews the changes of iron metabolism, iron-related mechanisms of brain injury, and the neuroprotective effect of iron chelator.

OBJECTIVETo identify the human hematopoietic stem cells from the human/goat xenogeneic model with molecular techniques.

METHODSDNA and total RNA were extracted from 11 transplanted goat peripheral blood cells. Human CD(34), GPA and SRY genes were amplified with PCR in these samples, and CD(34), GPA mRNA transcripts were detected using RT-PCR in 5 and 6 goat peripheral blood cells, respectively. Southern blot analysis was performed in 8 goat DNAs to detect the human specific alpha-satellite sequence. Meanwhile FISH was also performed to detect the human cells in goat blood with a probe of human Y chromosome.

RESULTSHuman CD(34) and GPA genes could be detected with PCR in all the 11 goats, and SRY gene did in 5 goats transplanted with hematopoietic stem cells derived from male human babies. Southern blot showed that human specific alpha-satellite sequence was present in 8 goats. By RT-PCR, human CD(34) mRNA was detected in 5 experimental goats, GPA mRNA was found in the other 6 experimental goats and FISH assay showed that some peripheral blood cells of the human/goat xenogeneic model were positive.

CONCLUSIONExistence of human cells in the recipient goats was identified by molecular detection, which was feasible for the examination of human/goat xenogeneic models.

Animals ; Antigens, CD34 ; genetics ; Blotting, Southern ; Female ; Genes, sry ; genetics ; Glycophorin ; genetics ; Goats ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Polymerase Chain Reaction ; Transplantation Chimera ; genetics ; Transplantation, Heterologous

As an effective vehicle for bio-research and for gene therapy, Lentiviral Vector (LV) has been drawn large attention in recent years. However, transcriptional read-through limits its application. In order to understand the extend of LV read-through in chromosome, a reliable method to assess transcriptional read-through rate is needed. Here, we report the method as follows: 293T cells were transfected with the lentiviral transfer vectors which borne with two LTRs at its two ends in order to mimic the state of "proviral vectors" in chromosome. Using the primers specific for 3'U5 and 3'U3, read-through and total transcripts were reverse transcribed, respectively. These two cDNAs were quantified by realtime PCR using the primers and probe specific for 5'end of 3'U3. Read-through rate was then calculated by the division of the two. Meanwhile, read-through product of green fluorescence protein was also analyzed by Fluorescence Activated Cell Sorter. They both reciprocally proved the principal and confirmed that self-inactivated LV appeared higher read-through rate than the wild type one. The method described in this article, therefore, provides a useful technique to study how to reduce read-through rate, and improve the bio-safety of LV.
Flow Cytometry ; Genetic Therapy ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; HEK293 Cells ; Humans ; Lentivirus ; Transfection

Objective To investigate the appropriate age of primary and secondary school students for cardiopulmonary resuscitation (CPR) training.Methods A total of 437 students aged 9-15 years at 3 to 6 grade in the primary schools or 1 to 2 grade in the secondary schools were selected from 2 Yuyao primary and secondary schools by stratified random sampling between March 2017 and January 2018.The numbers of students with the age of 9,10,11,12,13,14 and 15 y were 61,62,66,64,63,63 and 58,respectively.All students received chest compression training provided by Yuyao emergency department People's Hospital according to the 2015 Cardiopulmonary Resuscitation Guidelines.The training included 30 min theoretic teaching and 6 min practice in the simulator.The quality of chest compression performed by students was assessed;the depth,rate,position and retention of chest compression were recorded.Results The mean depth of chest compression in the students aged 9-15 years was 3.8,4.1,4.6,5.1,5.2,5.6 and 5.6 cm,respectively;the accuracy rate was 24.6％(14/61),25.8％ (16/62),50.2％ (33/66),70.5％ (45/64),79.4％(50/63),88.9％(56/63) and 91.4(53/58),respectively.Compared with the students aged 9-11 years,the mean depth of chest compression was significantly increased and accuracy rate was significantly improved in the students aged 12-15 years (Compared with 9-y students,t=-8.936,-9.502,-10.640 and-11.370;x2=35.019,47.599,63.013 and 65.671;compared with 10-y students,t=-6.927,-8.179,-10.70 and-11.047;x2=24.977,35.967,50.916 and 52.727;compared with 1 1-y students,t=-3.095,-4.177,-6.785 and-6.995;x2=5.586,12.114,22.786 and 24.870;all P＜0.05).The mean rate of chest compression was 110-116/min and its accuracy rate was 86.4％-95.2％;the accuracy rate of chest compression position was 90.9％-96.8％ in all students,there were no significant differences among the 7 groups.The mean retention rate of chest compression in the 7 groups was 81.3％(122/150),67.3％(101/150),64.7％ (94/150),48.0％(72/150),48.7％(73/150),33.3％(50/150) and 27.3％(41/150),respectively.Compared with the students aged 9-11 years,the mean retention rate of chest compression was significantly decreased in the students aged 12-15 years (compared with the 9-y students,x2=36.472,35.179,70.64 and 119.92;compared with 10-y students,x2=11.483,10.728,34.682 and 72.150;compared with 11-y students,x2=6.528,5.927,25.855 and 59.11;all P＜0.05).Correlation analysis showed that the depth (r=0.96,0.89,0.91 and 0.86;P＜0.01) and retention rate (r=-0.99,-0.90,-0.93 and-0.86;all P＜0.01) of chest compression were significantly associated with the age,body weight,height and body mass index of students.Conclusion The students with an age of 12 years or more are able to effectively perform chest compression;thus,12 years and above might be the appropriate age for CPR training.

OBJECTIVE: To investigate the effects of exendin-4 on hepatic lipid metabolism, fibrosis and oxidative stress in mice with streptozotocin (STZ)-induced diabetes and explore the underlying mechanisms. METHODS: C57BL/6J mice were fed with high-fat diet (HFD) for 4 weeks and received intraperitoneal injections of 120 mg/kg STZ to induce diabetes. After successful modeling, the mice were randomized into diabetic control group and exendin-4 treatment group (DM+E4), and in the latter group, the mice were given a daily dose of 1 nmol/kg of exendin-4 for 8 weeks. The changes in the body weight (BW) and random blood glucose (RBG) in the mice were recorded. The mRNA expressions of the genes related with liver lipid metabolism, fibrosis and oxidative stress were analyzed using RT-PCR, and the structural changes of the liver tissues were observed with HE, Sirius red and oil red O staining; the expressions of TGF-β1, Nrf2 and HO-1 proteins in the liver tissues were detected using Western blotting. RESULTS: The diabetic mice showed significantly higher RBG levels and BW with obvious lipid deposition, fibrosis and oxidative stress in the liver as compared with the normal control mice ( < 0.001). Exendin-4 treatment of the diabetic mice did not significantly lessened liver lipid deposition but obviously reduced the levels of RBG and TG ( < 0.05), lowered the expression levels of liver fibrosis-related genes TGF-β, -SMA and Col-Ⅰ ( < 0.05), increased the expression levels of the antioxidant genes Nrf2, HO-1 and GPX4 ( < 0.01), and enhanced the protein expressions of Nrf2 and HO-1 in the liver tissues ( < 0.01). CONCLUSIONS Exendin-4 improves liver fibrosis and oxidative stress in diabetic mice by activating Nrf2/HO-1 pathway without significantly reducing liver lipid deposition.
Animals ; Diabetes Mellitus, Experimental ; Exenatide ; Liver ; Liver Cirrhosis ; Mice ; Mice, Inbred C57BL ; NF-E2-Related Factor 2 ; Oxidative Stress ; Streptozocin