AIM:To reconstruct the tissue engineering blood vessel by using acellular matrix of porcine thoracic aorta tissue as the scaffold, and by inoculability of vascular endothelial cells (EC) from human umbilical cord vein. METHODS: The porcine thoracic aorta was treated with 1% Triton X-100 for preparing acellular vessel matrix, and the mechanical characterization was in succession modified by freeze-drying and thermal cross-linking techniques. Meanwhile, the mechanics capability of the vessel was measured. The endothelial cells isolated from umbilical cord vein were seeded on the acellular matrix scaffolds by tissue culture in vitro. The structure of acellular matrix was analyzed by light microscopy and scanning electron microscopy. RESULTS: By treatment with 1% Triton X-100 for 84 h, the cells of thoracic aorta were fully come off and the three-dimensional structure of the matrix still remained. After modification by freeze-drying for 24 h and then thermal cross-linking under vacuum at 120 ℃ for 12 h, the tensile strength of the acellular matrix remarkable increased and reached the maximum breaking strength of 1.70 MPa. It was also showed that cultured endothelial cells grew on the surface of acellular matrix for 7 days and the typical structure of vessel-like intimal formation was observed under scanning electron microscopy. CONCLUSION: The acellular matrix and endothelial cells have favorable compatibility. The modified acellular matrix could be a good candidate scaffold for rebuilding the tissue engineering blood vessel.

Objective To investigate the effect of high concentration carbon dioxide preconditioning on lipid peroxidation during myocardial ischemia-reperfusion (I/R) in rabbits. Methods Twenty-four New Zealand white rabbits weighing 2.0-3.9 kg were randomly divided into 3 groups ( n = 8 each): sham operation group (group S) , I/R group, high concentration carbon dioxide preconditioning group (group H) . The amimals were tracheal intubated and mechanically ventilated. In groups S and I/R, fresh gas flow was set at 0.3 L/min (100% O2 ), respiratory rate 30-40 bpm and tidal volume IS ml/kg, and PETCO2 was maintained at 40-50 mm Hg for 30 min. In group H, fresh gas flow was set at 0.3 L/min (100% O2), respiratory rate 20-30 bpm and tidal volume 10 ml/kg, PETO2 was maintained at 75-85 mm Hg for 5 min, and then all the ventilatory parameters were adjusted to the same as those in groups S and I/R. Myocardial I/R was produced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 3 h reperfusion after preconditioning in groups I/R and H. The animals were sacrificed at the end of reperfusion and myocardial tissues obtained for determination of the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content and examination of the ultrastnicture of myocardium with the transmission electron microscope. Results The SOD activity was significantly lower, while MDA content higher in group I/R than in group S ( P ＜ 0.01) . The SOD activity was significantly higher, while MDA content lower in group H than in group I/R ( P ＜ 0.01) . The myocardial injury was attenuated in group H compared with group I/R. ConclusionHigh concentration carbon dioxide preconditioning can reduce myocardial I/R injury in rabbits through inhibiting lipid peroxidation.

Objective To investigate expression of the phosphatase and tensin homolog (PTEN) gene in breast cancer cell line and its effect on biologic characteristics.Methods The normal PTEN expression cell line MDA-MB-231 (M231) was used in this study.PTEN-shRNA plasmid was transected into M231 breast cancer cells to knock down the expression of PTEN.The changes of PTEN expression,proliferation,invasion and metastasis of PTEN knocked down cell were tested by RT-PCR,Western blot,CCK-8,scratch and Transwell.Results PTEN-shRNA was successfully transected into M231 cells.PTEN mRNA and protein expression was efficiently inhibited in M231-3001 cell lines than that in control group M231-scr(P ＜ 0.01),M231-3001 cell lines showed a greater capability of colony formation,migration and invasion than that in control group M231-scr (all P ＜ 0.05).Conclusions PTEN,as a suppression gene,its low expression can promote the proliferation,migration and invasion of breast cancer cells.

Objective:To investigate the changes of lung lymphocytes in OVA induced murine asthma model,and study the involvement of NK cells in this process.Methods:C57BL/6J(B6) mice were induced to develop asthma by intrapenitoneal injection of OVA with alum as adjuvant, and then inhalation of nebulized OVA. After collecting serum and Bronchoalveolar Lavage Fluid(BALF),IL-4 level was determined by ELISA. The kinetics of pulmonary lymphocyte recruitment and cytokine release were detected by flow cytometry.Results:IL-4 expression increased in BALF after OVA nebulization, while there was no significant difference in serum. IFN-?,IL-4+NK cells accumulation in lung parenchymal tissues,and exhibited an evident NK2 shift in mice with asthma.Conclusion:NK cell involved in OVA-induced mouse asthma and NK2 shift accompany with Th2, indicating that NK2 played an important role in this process.

Objective To explore the microRNA expression changes and related characteristics and analyze the corresponding miRNA target genes and their bioinformatics in colorectal cancer with liver metastasis.Methods The fresh specimens of primary CRC were collected in 10 patients during operation,with liver metastasis or without.The miRNA expression levels were compared by miRNA microarray between two groups.Then,target genes were identified using target prediction algorithms.The liver metastasis related miRNA-target networks and gene ontology (GO) bioinformatics analysis were further performed.Results A total of six dysregulated miRNAs were identified in colorectal cancer liver metastasis comparing with no metastasis,including 3 up-regulated miRNAs (miR-224,miR-1236,miR-622) and 3 downregulated miRNAs (miR-155,miR-342-5p,miR-363).miR-224 with most significantly up-regulation played regulation role not only with corresponding target-genes but also downstream genes.Conclusions As a core of the regulation networks,miR-224 can regulate the related gene functional groups simultaneously and asynchronously.It further implements the post-transcriptional regulation and plays a vital role in liver metastasis of colorectal cancer.

Objective: To investigate the function of the ATP-sensitive K+(KATP) channel activation on the protective effect of hypercarbonic acidosis preconditioning on rabbit myocardial cells. Methods: Thirty-two rabbits were randomly divided into 4 groups (n = 8 for each group): pseudo-operation group (group P), ischemia and reperfusion group(group IR), hypercarbonic acidosis group(group H) and hypercarbonic acidosis+ glybenzcyclamide group (group H+G). Animals were ventilated normally in group IR and group P, tidal volume 15 mL/kg, breathing rate 35 bpm .The PETCO_2 was maintained at the level of 40-50 mm Hg for 30 minutes. Animals received low-frequency, low volume ventilation in group H group H+G, tidal volume 10 ml/kg, breathing rate 25 bpm to achieve hypercarbonic acidosis. The target value of PETCO_2 was 75-85 mm Hg. This value was maintained for 5 minutes. The animals then were ventilated normally to make the PETCO_2 return to 40-50 mm Hg. Animals were injected with 0.3 mg/kg glybenzcyclamide 10min before achieving hypercarbonic acidosis with hypoventilation in group H+G. Animals received ligation of left anterior branch artery for 30 minutes and reperfusion for 180 minutes in each group except P group. The myocardial ischemia area, the myocardial infarction area and their ratios were calculated by the ismaeil methods. Results: The ratio of the myocardial infarction area to the myocardial ischemia was significantly less in group H than those of group IR and group H+G (P < 0.01). The value of the ratio was similar between group H+G and group IR(P > 0.05). Conclusion: Hypercarbonic acidosis preconditioning can protect the cardiomyocytes by activating the KATP channel.

We investigated and analyzed the first case of human infection with avian influenza A(H9N2) virus in Yunnan Province,China,so as to provide a better basis for preventing and controlling human infections with viruses of animal origin in the future.We carried out the field epidemiological survey among the patient,close contacts and the live poultry markets,detected and analyzed the samples from patient and the outdoor environment.Results showed that the 9-month-old boy was a case of human infection with avian influenza A(H9N2) virus with the history of live poultry markets exposure and the results of nucleic acid detection and virus isolation.There was a lot of contamination of the avian influenza virus in the live poultry markets.The second generation cases have not occurred.The monitoring of pneumonia of unknown etiology and influenza like cases in medical institutions is the important means to find timely cases of human infected with avian influenza.Regular disinfection and closing-down of live poultry markets are key measures to reduce the exposure opportunity.

Objective To investigate the mutation of hemagglutinin gene and amino acid variation of influenza B.Methods Influenza B virus was isolated from throat swab samples in sentinel surveillance of Yunnan province from 2009 to 2014.HA1 gene sequence analysis was applied to determine 12 randomly-selected strains of influenza B virus.The results were analyzed,MEGA software was used to do homology comparison and HA gene phylogenetic tree was established.Results Differences on the serotype and genotype identification of influenza strains were found and it might be caused by inadequate gene mutation accumulation.Amino acid variations were found in 3 important regions of antigenic determinants in HA1 protein:ring 120,ring 150 and ring 160.The amino acid variation of position 131 in ring 120 was N131K,and in position 137 was N137H.Two strains had P187S mutation in position 187.Conclusion There are some important variations in the hemagglutinin gene of influenza B strains in Yunnan Province,with some variations being the same as vaccine strains and some being not.

Purpose To investigate the expression of polo-like kinase 1 (Plk1) and Cyclin B1, p21WAF1in cervical carcinoma, and to determine the relationship between the expression of the three proteins and tumor clinicopathological features. Methods The expres-sion of Plk1, Cyclin B1 and p21WAF1 was detected in 102 cases of cervical carcinoma, 20 cases of (cervical intraepithelial neoplasia, CIN) , and 20 cases of nomal cervical tissues by the technique of tissue chip and immunohistochemical staining of EliVision. Statistical analyses of the data were performed with SPSS 19. 0 software. Results The positive rates of Plk1 in cervical carcinoma and CIN were 70. 5%, 55. 0%, respectively, which were significantly higher than normal cervical tissues (10%) (P<0. 01);The positive rates of Cyclin B1 in cervical carcinoma and CIN were 52. 9% and 30. 0%, respectively, which were significantly higher than normal cervical tissues (10%)(P <0.01); The positive rates of p21WAF1 in cervical carcinoma and CIN were 23.5% and 10.0%, respectively, which were significantly higher than normal cervical tissues ( 0 ) ( P<0. 01 ) . There were no significant differences between cervical carcinoma and CIN in the positive rates of Plk1, Cyclin B1 and p21WAF1. The expression of Plk1 was associated with the depth of carci-noma invasion (P<0. 05), that of Cyclin B1 was associated with lymph node metastases and the depth of carcinoma invasion (P<0. 05)and that of p21WAF1 in cervical carcinoma was associated with histological grade (P<0. 05). In cervical carcinoma, the expres-sion of Plk1 was positively correlated with Cyclin B1 (rs =0. 297, P=0. 002) and negatively correlated with p21WAF1(rs = -0. 403, P<0. 001). Conclusion The expression of Plk1, Cyclin B1 and p21WAF1 is involved in the occurrence and development of cervical carcinoma, and the former two are also related with prognosis of cervical carcinoma. The combination of the three would provide a new target for clinical treatment.

This study observed the protective effect of hypercapnic acidosis preconditioning on rabbit heart suffered from ischemia-reperfusion injury. Hypercapnic acidosis was established in animals with mechanical hypoventilation before ischemia-reperfusion. Thirty-two rabbits were randomly divided into 4 groups, with each having 8 animals in term of the degree of acidification: hypercapnic acidosis group A (group A), hypercapnic acidosis group B (group B), hypercapnic acidosis group C (group C), ischemia and reperfusion group (group IR). Animals in group IR were ventilated normally (tidal volume: 15 mL/kg, breathing rate 35 bpm). The PETCO(2) was maintained at the level of 40-50 mmHg for 30 min. Animals in groups A, B, C received low-frequency, low-volume ventilation to achieve hypercarbonic acidosis and the target levels of PETCO(2) were 75-85,65-75, 55-65 mmHg, respectively, with levels being maintained for 5 min. The animals then were ventilated normally to lower PETCO(2) to 40-50 mmHg. The left anterior branch artery of all the animals was ligated for 30 min and reperfused for 180 min. Then the infarct size was calculated. The cardiomyocytes were morphologically observed and ECG and hemodynamics were monitored on continuous basis. Acid-base balance was measured during procedure. Our results showed that the infarct size was (48.5+/-11.5)% of the risk area in the control group and (42.4+/-7.9)% in group C (P>0.05). Mean infarct size was significantly smaller in group B (34.5%+/-9.4%) (P<0.05 vs control group) and group A (31.0%+/-9.1%) (P<0.01 vs control group). It is concluded that HA-preconditioning can effectively protect the myocardium.
Acidosis, Respiratory/*physiopathology ; Hypercapnia/*physiopathology ; Ischemic Preconditioning, Myocardial/*methods ; Myocardial Reperfusion Injury/*prevention & control ; Random Allocation