Objective To summarize and analyze the primary data from China Heart Transplant Registry in 2013 in order to explore the current status of heart transplantation in Chink Method Seventeen transplant centers have conducted 231 cases of heart transplantation in the year 2013.We analyzed the preoperative risk factors which impacted the survival of heart transplant recipients by using univariate and multivariate logistic regression analysis.And long-term survival of the 398 heart recipients from Beijing Fu Wai Hospital,who were followed up from June 2004 to December 2013 (follow-up rate was 100 ％) was calculated by the Kaplan-Meier method.The risk factors for long-term survival of heart transplant recipients were calculated by the COX survival analysis methods.Result The mean hospital stay of the 231 recipients was 24 days.There were 25 cases who died in hospital,and the total mortality was 10.8 ％ (25/231),while the average in-hospital mortality was 2.2 ％ in the three largest centers.By using the univariate and multivariate logistic regression analysis,we found that recipients positive PRA was the only preoperative risk factor which impacted the in-hospital mortality.(OR=12.435,95％ CI 1.369～ 112.938,P =0.025).The median follow-up time of the 398 heart transplant recipients from Beijing Fu Wai Hospital was 1383 days,and the 1-,3-,5-and 7-year survival rate was 94.7％,91.6％,88.0％ and 82.6％ respectively.The univariate and multivariate COX regression analysis revealed that preoperative primary diseases (coronary heart disease vs.cardiomyopathy,P ＜ 0.01),the history of chronic obstructive pulmonary disease (P ＜ 0.01),the preoperative total bilirubin level (P＜0.05) and serum creatinine level (P＜ 0.01) were risk factors affecting long-term postoperative survival.Conclusion In China the number of heart transplants has increased in 2013.The survival rate of recipients is high in large heart transplant centers.The integrity of registration data needs to be further improved in some centers.

Objective:To construct an in vitro reconstitution system for inverse autotransporters in order to further investigate their translocation mechanism. Methods:Intimin from Escherichi coli was used as a model substrate. Spheroplasts were prepared from Escherichi coli strains overexpressing Intimin to induce the expression of Intimin. Recombinant β-barrel assembly machinery (BAM) complex was obtained and purified, and then proteoliposomes containing BAM were prepared. Following the digestion with proteinase K, the translocation was detected by SDS-PAGE. Results:Spheroplasts were induced to express Intimin, and then BAM-containing proteoliposomes were added to the system. Compared with control and liposomes groups, the experimental group showed that Intimin was resistant to proteinase K treatment, indicating that Intimin was successfully translocated.Conclusions:The translocation of Intimin required the participation of BAM complex. An in vitro reconstitution system for inverse autotransporters was constructed in this study, providing a method to study the translocation mechanism of inverse autotransporters.

Objective:To construct a surface display system containing various lengths of the Ag43 passenger domain for an optimal bacterial surface display of foreign protein HPV16L1.Methods:(1) Ag43 gene sequences of different lengths were inserted into pET22b vector to construct four Ag43 surface display vectors (Ag43/138, Ag43/551, Ag43/552 and Ag43/700) using PCR and subcloning strategy. (2) The generation of four HPV16L1-Ag43 fusion constructs was completed by PCR and subcloning methods. (3) HPV16L1-Ag43 fusion proteins were expressed and analyzed by SDS-PAGE. (4) The surface exposure of HPV-16L1 was verified using trypsin digestion.Results:PCR analysis and sequencing results showed that Ag43 surface display vectors and HPV16L1-Ag43 fusions were constructed successfully. SDS-PAGE showed that the expression of HPV16L1-Ag43 fusion proteins could be induced with 0.2 mmol/L IPTG and the protein content was reduced after the cells were treated with trypsin, especially the content of Ag43/700-HPV16L1 that showed a drastic reduction.Conclusions:The Ag43 surface display system was successfully constructed and could be used for a successful display of HPV16L1. This study also showed that Ag43/700 comprising only the α-helix and the β-barrel of Ag43 provided an optimal surface display for HPV16L1.