BACKGROUND:Establishing an animal model of lung transplantation has operational significance to the development of clinical lung transplantation.
OBJECTIVE:To analyze and summarize animal selection, model establishment method, points to note and mechanism of lung ischemia-reperfusion injury and pulmonary immune rejection in the establishment of animal models as preclinical research of lung transplantation.
METHODS:A computer-based retrieval of CNKI and PubMed database from January 1982 to September 2013 was performed for literature related to lung transplantation. The key words were“lung transplantation, animal models, reperfusion injury”in English and Chinese, respectively. After eliminating duplicate and obsolete literature,50 articles were included for further analysis.
RESULTS AND CONCLUSION:Currently, animal experiments about al ogeneic lung transplantation are
common, which are of significance for clinical lung transplantation. Single lung transplantation is mainly seen in animal models. The commonly used lung transplantation models include single lung orthotopic murine model, rabbit orthotopic lung transplantation, canine and porcine orthotopic left lung transplantation, canine bilateral sequential lung pulmonary transplantation, autologous pig lung transplant model, rabbit lung ischemia-reperfusion model. Because the anatomical and physiological features are similar to humans, pigs are becoming the preferred choice for large animal experiments. Animal experiments and clinical studies have shown that lung ischemia-reperfusion injury in lung transplantation represents a biphasic pattern:early occurrence of lung ischemia-reperfusion injury (24 hours after infusion) is related to lung donors, and the late occurrence mainly depends on receptors. Its pathophysiology runs through the whole process of donor lung resection, preservation and reperfusion as wel as postoperative management. Immune rejection is a complex immune response induced by identifying the donor cellsurface histocompatibility antigen, and activated T lymphocytes play a crucial role in the immune response in organ transplantation.

Objective: To investigate the rational use of antibiotics in the elderly patients with cerebral infarction complicated with pulmonary infection.Methods: A retrospective investigation was used to analyze the drug resistance of 86 cases of elderly patients with cerebral infarction complicated with pulmonary infection.The data were collected from the consultation records of clinical pharmacists in Xuanwu hospital from February 2013 to November 2015.Results: The pathogenic bacteria mainly distributed to Acinetobacter baumannii and Pseudomonas aeruginosa, and the drug resistance rate was 48.89%.After the consultation, the rate of combination therapy increased from 20.23% to 61.63%.After using sensitive antibiotics, such as cefperazone-sulbactam,piperacillin-sulbactar and amikacin, the infectious indices recovered significantly (P＜0.05).The case proportion with BUN/SrCr＞20 in 27 cases complicated with impaired renal function was 62.9%, and although the patients didn''t use antibiotics with reduced doses, the indices of infection and renal function improved in a certain degree (P＜0.05).The case proportion with ALB and PAB decreasing was 53.49%, and after the consultation, all the infectious indices decreased and protein levels increased (P＜0.05).Conclusion: More attention should be paid to the elderly patients with cerebral infarction complicated with pulmonary infection, and sensitive antibiotics with single or combination drug therapy should be used according to the distribution of resistant bacteria.For the renal failure patients, the kind of renal failure should be clear, and then the appropriate dosage of antibiotics should be adjusted.For the patients with malnutrition, adequate protein intake contributes to the recovery of infection.The clinical symptoms and laboratory indices should be integrated to effectively develop rational treatment for the elderly patients with cerebral infarction complicated with pulmonary infection.

Comet Assay

Objective:To mutate the codons which code the lysine at loci ? 99 ,? 82 in human hemoglobin gene to the codons which code the cystein.Methods:The objective gene to be mutated was cloned into pAlter Ex2 plasmis and then was mutated with the site directed mutagenesis mediated by oligonucleotide.Results:The sequencing of clones obtained by preliminary screening showed that the anticipated results were reached without any unexpected mutation.Conclusion:The results lay a foundation for the further development of research on expression and purification of recombinated mutant human hemoglobin gene.

Objective To make an attempt to express serialy the human ?,? globin in Escherichia Coli in order to carry out the research on blood substitute based on the genetic recombination of hemoglobin.Methods The routine molecular biology technology was used.Results The serial genetic fragment of ?,? globins containing the hemoglobin was amplified by PCR.The genetic sequencing showed that there was no unexpected mutation.And then,the serial genes were cloned into the pBV220 expression carrier.Undergoing thermal inducement,the expression product could reach about 20% of total bacterial protein.The expression product assumed the form of inclusion body,and it was vertified by Western Blotting.Conclusion This much can lay the solid foundations for further launching the research on expression and pruification of recombinant human hemoglobin genes.

Objective To design and identify small interfering RNA (siRNA) targeting tumor necrosis factor‐α (TNF‐α) ex‐pression ,siRNAs were electroporated into synovial cells of Human to screen those which can effectively suppress TNF‐α expres‐sion .Methods Three TNF‐α specific double stranded siRNA were designed targeting different regions of TNF‐α mRNA(compared with negative control group) .RT‐PCR and Elisa were applied to detect TNF‐α ,mRNA expression and the secretion of TNF‐α in cell supernatant ,respectively .Results Two of the three customized TNF‐α siRNA could inhibit the expression of TNF‐α mRNA in synovial cells of Human (P< 0 .01) .At the same time ,the secretion of TNF‐α decreased in cell supernatant ,the difference was sig‐nificant statistically compared with the control group(P< 0 .01) .Conclusion TNF‐α siRNA can be successfully designed and syn‐thesized ,which can specifically and effectively suppress TNF‐α mRNA expression .

Objectives To study the effects of effective constituent of Cortex Moutan-Paeonol and Paeoniflorin on myocardial ischemia and myocardial infarction in experimental canines.Methods The coronary artery ligation was employed to establish the model of canines with acute myocardial ischemia and myocardial infarction.The extent of myocardial ischemia was detected by epicardial-electrocardiogram and the extent of infarction was determined by N-BT staining method.Results The adminstration of Paeonol and Paeoniflorin reduced the myocardial ischemia level(P

Objective To evaluate the feasibility, safety and efficacy of selective portal vein embolization (SPVE) in treatment of liver tumor in rats and to provide the groundwork for its future clinical applications. Methods 24 healthy rats underwent the embolization. Pre and post SPVE portogram and liver chemical profile were obtained. Four rats were sacrificed at 10 min, 7,14, 21 and 28 days respectively following follow up portography. The liver, heart, lungs and kidneys were examined macroscopically and microscopically. Fifteen rats implanted with Walker 256 tumor sized from 3 to 10 mm in liver were scanned with MRI and portography pre SPVE taken. Post SPVE 3 rats were examined with MRI for each group at the same interval as above and the lives were examined microscopically. Results (1) The blood flow to the target portal branches were immediately halted after SPVE. These vessels remained occluded without collateral formation up to 28 days. (2) The liver indexes and BUN level increased after embolization, but returned to normal within 21 d. Macroscopic and microscopic changes were not found in the heart, lungs or kidneys. (3) In the healthy rats, the affected segment was atrophic and the remaining liver underwent compensatory hypertrophy. Histologic examination revealed that the targeted portal veins were coagulated, the endothelium were degenerated and the local hepatocytes were necrotic after embolization. (4) In the rats with implanted liver tumor, the affected segment including the tumor was necrotic and atrophic. The tumors were completely necrotic, and no viable tumor cell was seen under microscope in 12 among the 15 rats. Three tumors 10 mm in diameter were not completely necrotic. Part of tumor cells were still alive and infiltrated into the surrounding liver. Conclusion SPVE with ethanol is effective in the treatment of small liver tumor in rats. However,in case of bigger tumors involving several segments, SPVE should be combined with other treatment.

Objective To investigate pathogene city, replication and expression of hepatitis G virus (HGV) in rhesus monkey infected with HGV RNA genome or HGV RNA positive sera. Methods A full length cDNA clone of HGV was constructed. Rhesus monkey BY1 was inoculated intrahepatically with genomic RNA from this HGV clone resulted in viral replication. HGV RNA positive sera from BY1 were intravenously inoculated into rhesus monkeys BM1, and sera from BM1 were intravenously inoculated into BB1 in series. Sera were collected weekly or bi weekly and liver biopsies were performed regularly. RT PCR, in situ hybridization and immunological, serological, histological assays were carried out to study the infectivity and pathogenecity of HGV. Results The serological and pathological results indicated that all of the 3 rhesus monkeys developed HGV viremia and had slightly elevated alanine transaminase levels (up to 418 IU/ L) during the period of experiment. HGV RNA became positive at the 3 rd , 8 th and 3 rd week post inoculation in the animals BY1, BM1 and BB1 respectively, and existed up to 21 weeks. The histology, immunohistochemnistry, and in situ hybridization in the liver tissues of the inoculated animals also showed that there was a mild hepatitis with HGV E2 expression in cytoplasm of hepatocytes. RT PCR and quantitative PCR showed that HGV could replicate in liver.Conclusions The genomic RNA from HGV full length cDNA is infective to the rhesus monkeys resulting in mild hepatitis. Infection and the transmission of the HGV in the rhesus monkey provide an appropriate animal model for the study of HGV.

Objective To review the clinical features of chronic daily headache (CDH).Methods The clinical data of 128 patients with chronic daily headache,including general condition,characteristics of headache,concomitant symptom and disability were analyzed retrospectively.The features of primary chronic daily headache (PCDH) and medication over-dose headache (MOH) were compared.Results Among 128 cases females accounted for 79.7％ with an average age of 45.2 years and 88 patients were associated with drug overdose.The symptoms included nausea (68/128),photophobia (75/128),phonophobia (102/ 128),depression (77/128) and irritability (93/128),sleep disorders (94/128),dizziness (75/128),emotional irritability(58/128) and depression(21/128).The migraine disability assessment questionnaire and headache impact test-6 scores showed that disability was resulted from the severe degree of headache in 62.2％ (51/82) and 73.2％ (82/112) of CDH patients respectively.Compared with PCDH patients,the MOH patients had older age (t =2.59,P =0.011),longer duration (t =2.48,P =0.015) and severer degree of headache(t =5.58,P =0.018),and chronic migraine (t =11.95.P =0.001) was the most common primary headache type.Conclusions Most CDH patients are middle-aged women,with moderate to severe pain,usually complicated with depression,dysphoria and asomnia.Chronic daily headache patients are commonly associated with drug overdose.