BACKGROUND: The method and effect of construction of tissue-engineering heart valve scaffold with cell detergents are different, and the method of detergent combining with the trypsase and nucleese is more suitable than others. OBJECTIVE: To investigate the effect of different detergents (sodium deexycholate, sodium dodecylsulfate, and triton) combined with trypsase on decellularized porcine heart valve and the influence on the accellular scaffold. METHODS: Porcine aortic valve leaflets were sterilized by antibiotics for 12 hours, maintained in the solution of the trysin and the EDTA for 12 hours, and treated with sodium deexycholate, sodium dodecylsulfate, and triton. Finally, the sample was dip in nucleese solution for 12 hours to remove endothelial cells and interstitial cells. HE staining was used to detect whether the endothelial cells were removed completely, Masson staining was used to evaluate damage level of collagen fiber and elastic fiber, and electronic scanning was used to observe the microstructure. RESULTS AND CONCLUSION: All the three detergents completely removed the endothelial ceils; however, the effect of sodium decxycholate on collagen fiber and elastic fiber was light, and then sodium dodecylsulfate and triton. This suggested that the method of DOA combining with the enzyme digestion was a suitable technique to construct tissue-engineering heart valve scaffold.

Objective To study the roles of protein kinase C (PKC) in the signal transduction pathway of LPS induced nuclear transcription factor (NF) ?B activation of alveolar macrophages(AMs). Methods NF ?B level in nuclear protein extraction of AMs was detected by sandwich ELISA. Intranuclear translocation of NF ?B was observed by immunocytochemical staining. Results LPS could induce NF ?B activation of AMs in time and dose dependent manners. Immunohistochemical staining revealed intranuclear translocation following LPS induction. Specific calphostin C (Cal C) and pyrrolidine dithiocarbamate (PDTC) could inhibit LPS induced NF ?B activation. Conclusion PKC, as a up stream messenger, is involved in the signal transduction pathway of LPS induced NF ?B activation.

Objective To explore the expression of interleukin-19 (IL-19)and to investigate the relationship between IL-19 and type 2 diabetes mellitus9 (T2DM)with angiopathy.Methods IL-19,FBG,FINS and HbA1c were measured in 120 T2DM (including 32 patients with macroangiopathy,52 patients with microangiopathy and 36 without angiopathy)and 50 healthy subjects,the results were compared.Results Compared with control group,the level of IL-19 in T2DM (41.9 ± 11.9pg/ml vs 16.2±8.5pg/ml)was significantly higher than those of healthy subjects,and the difference was signifecant (t=7.56,P<0.05).The levels of IL-19 in T2DM with macroangiopathy or microangiopathy increased significantly in com-parison with those without angiopathy (t=4.57,3.26;all P<0.05),and closely correlated with the number of angiopathy. The level of IL-19 in T2DM was significantly positive correlated with HOMA-IR and HbA1c (r=0.523 and 0.491,P<0.01).Conclusion IL-19 level was significantly increased in patients with T2DM,and was closely related with HbA1c,insu-lin resistance,IL-19 may play acertain role in the pathogenesis of in T2DM with vascular complications.

OBJECTIVE To investigate the prevalence of drug-resistant genes in seventeen strains of all-resistant Acinetobacter baumannii.METHODS Microdilute tests were performed to detect the susceptibility of 17 A.baumannii strains to 16 kinds of antimicrobial agents and antimicrobial-resistant genes were detected by PCR methods.RESULTS Seventeen A.baumannii strains showed all-drug resistance.Genes of TEM-1,OXA-23,OXA-27,gyrA and AmpC were detected in all 17 strains of A.baumannii.The positive rates of aacC31 and PER-1 genes were 11.8% and 52.9%,respectively.CONCLUSIONS A.baumannii with multi-resistant genes of our hospital carries TEM-1,OXA-23,OXA-27,gyrA,AmpC,aacC1 and PER-1.

Drug-induced immune hemolytic anemia has complicated manifestations and pathogenesis, and therefore clinicians should know its etiology and pathogenesis for safe medication. In this paper, we made a discuss on the etiology and pathogenesis of drug-induced immune hemolytic anemia from both traditional Chinese and western medicine viewpoint hoping to provide references for clinical physicians.

A neonatal girl with overextended knees admitted to NICU of our hospital was diagnosed as cri du chat (cat cry) syndrome.We collected 34 cases of cri du chat reported in journals since 2000,the clinical features of total 35 cases were retrospectively analyzed.Among 35 cases 12 were boys and 23 girls.The most common clinical manifestations were characteristic face features(100％),difficult feeding(100％) and typical sound of cry(94％).The main complains at hospital visit were typical cry,difficult feeding and cyanosis in the neonatal period,while in childhood period were recurrent respiratory infection,developmental retardation and other abnormalities.Most cases were diagnosed in the neonatal phase,while 85.3％ were in the first year.The diagnosis was based on karyotype analysis; chromosome 5 short arm deletion (5P-) was the most significant genetic variation and clinical features were associated with the position of deletion.

Objective To explore the expressions and clinical value of angiopoietin like protein 1 (Ang-1)and angiopoietin like protein 2 (Ang-2)in patients with gastric cancer.Methods 65 patients with gastric cancer and 50 healthy subjects for con-trol group were collected from June in 2012 to December in 2014.The level of Ang-1 and Ang-2 were determined by enzyme-linked immunosorbent assay (ELISA).The results were compared.Results Compared with control group (19.8±2.1 μg/L),the level of Ang-1 in patients with gastric cancer (19.8±2.1 μg/L)was no statistics difference (P >0.05),but Ang-2 (2.3±0.8 μg/L)was significantly higher than those of healthy subjects (0.8±0.2 μg/L,t=2.50,P <0.01).There was no significant difference of Ang-2 level in different pathological types of gastric cancer (P >0.05).The level of Ang-2 in TNM stage Ⅲ~Ⅳ (2.6±0.5 μg/L)was obviously higher than that in stage.Ⅰ~Ⅱ (1.6±0.4 μg/L).Ang-2 levels were signif-icantly higher in patients with lymph node metastasis (2.7±0.5 μg/L)or postoperative recurrence after one year (2.0±0.6μg/L)than those without lymph node metastasis (1.6 ± 0.5 μg/L)or postoperative recurrence (1.2 ± 0.5 μg/L,P <0.05).The level of Ang-2 in gastric cancer was significantly positive correlated with tumor stage and lymph node metastasis (r=0.31 and 0.33 respectively,P <0.01).Conclusion Ang-2 level was significantly increased in gastric cancer.Its level was significantly correlated with tumor stage,lymph node metastasis and prognosis,which has important application value for the cancer progression,clinical observation and prognosis evaluation of gastric cancer.

AIM: To explore the effect of complement on the cerebral ischemia/reperfusion injury in rat and the protection by sCR1-SCR15-18. METHODS: 75 male SD rats were randomly divided into three groups: sham operation group (SO, n=15), middle cerebral artery occlusion and reperfusion (MCAO) without treatment group (I/R, n=30); MCAO treated with sCR1-SCR15-18 group (sCR1-SCR15-18, n=30). After the MCAO for 2 h, then reperfusion for 24 h, the scores of neural behavioral functional deficits were determined. Infarction area was measured by TTC staining. Activity of MPO in cerebral cortex was detected. C3b deposition and pathological change were observed by immunohistochemial staining and HE staining, respectively. RESULTS: After reperfusion for 24 h, the neurological deficits score, infarction area and activity of MPO in sCR1-SCR15-18 group were decreased compared to I/R group. In sCR1-SCR15-18 group, C3b deposition in ischemic area was decreased and pathological injury was improved compared to I/R group. CONCLUSION: Complement plays a role in cerebral ischemia-reperfusion injury and sCR1-SCR15-18 exerts a protective effect by inhibiting the excessive activation of complement.

Objective To express the short consensus repeat 15-18 (SCR15-18) domain of human complement receptor 1 (CR1) in Pichia pastoris as a secreted protein in order to found a base for large-scale industry fermentation. Methods The gene fragment of CR1-SCR15-18 was amplified by PCR from plasmid pET32a-sCR1-SCR15-18. The obtained sequence was subcloned into Pichia pastoris secretory expression vector pPIC9. After identification the recombinant plasmid pPIC9-CR1-SCR15-18 was electrotransported into the yeast. Positive clones were identified using colony PCR. Positive recombinants were fermented in shake flaskes and induced with methanol. The recombinant proteins were identified with SDS-PAGE and Western blot analysis. After the recombinant protein was purified by Ni-NTA agarose metal chelate affinity chromatography, inhibiting complement hemolysis testing was used to detect the biological activity. Results Recombinant expression plasmid pPIC9-CR1-SCR15-18 was successfully constructed. SDS-PAGE and Western blot analysis revealed that the target gene was successfully expressed in the yeast and the recombinant protein was successful secreted in the culture supernatant. After purification, the protein inhibited complement hemolysis in vitro. Conclusion CR1-SCR15-18 is successful expressed in Pichia Pastoris with high activity of inhibiting complement hemolysis.

Objective To construct and express a small complement receptor type 1 (CR1) deriv-ative which contained two functional domains. Methods Total RNA was isolated from the peripheral blood mononuclear cell. The functional fragment Ⅰ of CR1 was amplified using the RT-PCR. The functional frag-ment Ⅱ was amplified with the plasmid of pET-32a-CR1-SCR15-18 as template which had been already con-structed in our laboratory. Then the chimeric gene that contained the two fragments was constructed with spli-cing overlap extention PCR. The chimeric gene was then inserted into the plasmid of pET-32a (+) and transformed into E. coli Rosetta(DE3). The inserted gene was verified by enzyme digestion and DNA se-quencing. After induced by IPTG, the expressed protein was analyzed by SDS-PAGE and Western blot. Then the fusion protein was purified by Ni-NTA affinity column and renatured through dialysis. The comple-ment inhibition activity was determined by CH50 method. Results The chimeric gene was successfully cloned into pET-32a(+). The result of SDS-PAGE and Western blot confirmed the expressed protein and showed that the molecule mass(Mr) of the expressed protein was 63×103. The purity of the recombinant protein was up to 92% after Ni-NTA column affinity chromatography. The bioactivity assay showed the fusion protein had a concentration-dependent complement inhibition activity within the concentration range of 0-200 μg/ml. Conclusion The two functional domains contained small CR1 derivative was successfully construc-ted and expressed in E. coli Rosetta. The fusion protein had a relative high bioactivity, providing a basis for further function experiment in vivo.