Aluminum Compounds ; toxicity ; Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cells, Cultured ; Cerebral Cortex ; cytology ; drug effects ; Chlorides ; toxicity ; Dose-Response Relationship, Drug ; Neurons ; cytology ; drug effects ; Rats ; Rats, Wistar

[Objective] To explore whether necroptosis is involved in the mechanism of lung injury induced by intestinal ischemia-reperfusion.[Method] Thirty-two healthy male Sprague-Dawley rats were randomly assigned into 4 groups (n--8):sham operation group (sham group),isehemia/ reperfusion group (I/R group),necroptosis inhibitor necrostatin-1 group (Nec-1 group) and solvent dimethyl sulfoxide (DMSO) group (DMSO group).Model of intestinal I/R injury was produced by clamping the superior mesenteric artery for 1.5 h followed by 6 h reperfusion in rats.Necrostatin-1 1.0 mg/kg was administered 30 min before occlusion in Nec1 group,while the equal volume of DMSO was given instead in DMSO group.The rats were sacrificed at 6 h of reperfusion and the lung tissues were removed for measurement of wet-dry ratio and microscopic examination and scored.The expression of receptor-interacting protein 1 (RIP1) and receptor-interacting protein 3 (RIP3) in lung tissues was detected using Western-blot and immunohistochemistry.[Result] Compared with sham group,lung morphology score and wet/dry ratio in I/R,DMSO group raised (P ＜ 0.05).Lung morphology score and wet/dry ratio statistically declined in Nec-1 group compared with I/R and DMSO group (P ＜ 0.05),while there was no statistical difference of wet/dry ratio between sham group and Nec-1 group (P ＞ 0.05).As the result of westernblot and immunohistochemistry showed,the expression of RIP1 and RIP3 was up-regulated in I/R group and DMSO group (P ＜0.05),which was inhibited by Nec-1 in Nec-1 group (P ＜ 0.05).[Conclusion] Necroptosis is involved in the mechanism of lung injury induced by intestinal ischemia-reperfusion,and Nec-1,the special inhibitor of RIP1,can reduce the injury.

AIM: Recent studies showed that stem cells could replace injured cardiomyocyte and increase the number of functional cardiomyocytes. Researching the pertinent literature in China Journal Full-text Database (CJFD) published between 2005 and 2008 indicated that the researches on stem cell transplantation in the treatment of primary dilated cardiomyopathy were few. This study investigated the therapeutic efficacy and security of peripheral blood stem cell transplantation in coronary artery in treatment of dilated cardiomyopathy and its effects on left ventricular function. METHODS: Forty-two patients with dilated cardiomyopathy between December 2006 and September 2007 were enrolled at the Department of Cardiology of First People's Hospital of Yunnan, including twenty-eight males and fourteen females, averagely aged (56?3) years. Inclusive criteria: patients with less than 65 years, left ventricular enlargement, and left ventricular ejection fraction (LVEF) ≤ 45%, and without coronary artery disease after coronary arteriongraphy. Informed consents were obtained from patients. Patients were divided into stem cell transplantation group (n=15) and control group (n=27) on the basis of whether being treated by stem cell transplantation. Patients in the stem cell transplantation group were consecutively administered granulocyte colony-stimulating factor (G-CSF) by hypodermic injection to stimulate bone marrow stem cells themselves based on conventional treatment for five days. Peripheral blood stem cell suspension was disassociated on the 6th day, and the collected suspension was injected into left anterior descending branch over the wire saccule tube for autologous peripheral blood stem cell transplantation. Patients in the control group were administered by conventional treatment of dilated cardiomyopathy. Security and adverse reaction were observed during the mobilization, collection and returning injection of peripheral blood stem cell by coronary artery. Morphous, cardiac function and motion index of left ventricle wall were evaluated using ultrasoundcardiogram before and 3 months after transplantation. Survival rate and incidence rate of heart incidents were compared. RESULTS: Three months after stem cell transplantation in coronary artery, there were a significant decrease in cardiac end-systolic volume (ESV), cardiac end-diastolic volume (EDV) and motion index of left ventricle wall, but a significant increase in LVEF(P

AIM: To study the protection of extract of Radix Atragali composite against acute hepatic injury. METHODS: Fed with the extract of Radix Atragali composite, m ice were injected with D-galactosamine intraperitoneally (800 mg/kg) and rats were i njected with carbon tetrachloride hypodermically (5 mL/kg) to induce acute hepat ic injury on the 8th day. ALT, AST and bilirubin in serum were examined. Patholo gical changes of liver tissue were observed. RESULTS: Compared with model group, activities of ALT and AST, c oncentrations of bilirubin were markedly decreased and pathological scores also showed that degeneration and necrosis of hepatic cell were lighter in the the ex tract of Radix Atragali composite treatment group. CONCLUSION: The extract of Radix Atragali composite attenuat es hepatic injury induced by D-galactosamine or carbon tetrachloride.

Objective To investigate the protective effect of endotoxin pretreatment on hippocampal neurons in rat forebrain following ischemia reperfusion and its possible mechanism. Methods Rat forebrain ischemia reperfusion model was used. The effects of endotoxin pretreatment on the changes of superoxide dismutase (SOD), glutathione peroxidase (GSH PX) and malondialdehyde (MDA) activities and the neuron count in CA1 region were observed. Results Pretreament with endotoxin before cerebral ischemia enhanced the activities of SOD and GSH PX but decreased MDA level and the number of ischemic neurons in CA1 region. Conclusion Endotoxin pretreatment can protect the neurons in rat forebrain against ischemia reperfusion injury. The mechanism may be associated with the enhancement of endogenous antioxidant activity in central nervous system.

Characteristics and measurement principles of reference methods in clinical biochemistry were described.Implementation of reference systems is one of the most effective approaches to improve the accuracy and comparability of clinical laboratory test results.Reference methods are the key components of reference systems.Reference methods should have measurement uncertainties that meet the requirements of the intended use,and thus should be based on reliable measurement principles.For the well-defined biochemistry analytes,reference methods have been almost all based on instrumental analysis.Isotope dilution mass spectrometry (ID/MS) is considered most reliable and has been the major analytical principle of the reference methods.ID/MS analysis is accurate but expensive.Use of other validated instrumental analyses as reference measurement principles would be justified.

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

A BBB co-culture cell model consisting of rat brain microvascular endothelial cells (BMEC) and astrocytes (AS) was established to study the effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier (BBB) in vitro and in vivo. The barrier function of this model was evaluated by measuring the transendothelial resistance, phenol red permeability and BBB related protein expression. The permeability assay and western blot methods were performed to study the effects of Angelica dahurica coumarins on the BBB permeability and the expression of BBB related protein. The animal experiment protocols in this study were approved by the Animal Ethics Committee of Xi'an Jiaotong University (Animal Ethics No.: 2021-1329). The results showed that the established BMEC/AS co-culture model could be used to evaluate drug transport across BBB in vitro. After combined with Angelica dahurica coumarins, the transport capacity of puerarin was significantly increased in vitro and in vivo. Additionally, Angelica dahurica coumarins enhanced BBB permeability and inhibited the protein expression of P-glycoprotein (P-gp), zonula occludens-1 (ZO-1) and occludin. Angelica dahurica coumarins might increase BBB permeability by inhibiting the expression of P-gp and tight junction protein, thereby increasing the content of puerarin in brain tissue.

To improve students' experimental research skills,innovative consciousness,promote the construction of functional experimental center,we established series of “comprehensive,contrivable,innovative” experiments in medical students.