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Nowadays, personal privacy disclosure issues become increasingly prominent in the process of data publishing. Targeting at this issue, a secure blockchain-based personal privacy protection scheme is proposed. In this paper, firstly, the information is stored in cloud service, and its hash value is used for the index, which is encrypted and stored on the blockchain. Secondly, based on the principles of cryptography and the characters of blockchain technology, we propose our scheme and use it to provide an access control mechanism for personal privacy. It can make the personal privacy protection easier and more efficient. At last, through our analysis, this scheme can sufficiently protect personal privacy in using mobile devices and provide a safe and trustworthy information acquisition method for data mining. More importantly, this proposed scheme also can provide a reliable and unique source tracing for multimedia copyright protection in terms of multimedia security.

In this paper, a novel quantum blockchain scheme is proposed to optimize the security of blockchain. Firstly, we propose the definition of quantum blockchain and provide its construction in detail. Additionally, its advantages are also summarized in this work. Secondly, based on the quantum no-cloning theorem, we define a new type of cryptocurrency which we call it quantum coin. Meanwhile, we adopt quantum entanglement and DPoS to design a novel quantum blockchain scheme. At last, we analyze the security of this proposed scheme in terms of the secret keys and quantum coin. It is shown that some attacks, such as man-in-the-middle attack, double spending attack and state-estimation attack, can be resisted. More specifically, under quantum computing attacks, the scheme is also secure. To sum up, through the principles of quantum cryptography and DPoS, this novel quantum blockchain obviously provides better security and higher efficiency.

BACKGROUND: Revascularization for the treatment of coronary artery disease (CAD) is advancing rapidly and is used increasingly in old patients. This study aimed to compare the efficacy and safety of revascularization with drug therapy in CAD patients aged over 80 years at a real-world clinical setting. METHODS: A total of 501 CAD patients aged over 80 years were consecutively enrolled from January 2011 to January 2016 in Anzhen Hospital (Beijing, China), Capital Medical University. The patients were treated with percutaneous coronary intervention (PCI) (n=283), coronary artery bypass grafting (CABG) (n=106), or drug therapy (n=112). All-cause mortality, cardiovascular-related mortality, readmission rate, and Seattle Angina Questionnaire (SAQ) score were compared between the three treatment methods. RESULTS: A total of 411 patients (82.04%) were followed with a median duration of 25 months. All-cause mortality and cardiovascular-related mortality in the drug therapy group were significantly higher than the PCI and CABG groups (both P

Recently, some shortcomings have exposed over the application and management of digital technologies of gas production with deliquification in Sulige gas field. Through summing up and analysing the challenges, experts have improved and promoted the digitization and intelligence of gas production with deliquification technologies in Sulige gas field.

Background: Unfractionated heparin (UFH), despite its limitations, has been used as the primary anticoagulant alternative during the percutaneous coronary intervention (PCI). Some studies indicated that intravenous enoxaparin could be an effective and safe option. Our team used enoxaparin alone at one time according to the guidelines (Class IIA) and found a little catheter thrombosis during PCI. We recommend a new anticoagulation strategy using enoxaparin in combination with UFH. Enoxaparin has a more predictable anticoagulant response with no need of repeatedly monitoring anticoagulation during PCI. This retrospective study aimed to evaluate the efficacy and safety of using enoxaparin in combination with UFH in PCI patients with complex coronary artery disease. Methods: Between January 2015 and April 2017, 600 PCI patients who received intravenous UFH at an initial dose of 3000 U plus intravenous enoxaparin at a dose of 0.75 mg/kg (observation group) and 600 PCI patients who received UFH at a dose of 100 U/kg (control group) were consecutively included in this retrospective study. The endpoints were postoperative 48-h thrombolysis in myocardial infarction (TIMI) bleeding and transfusion and 30-day and 1-year major adverse cardio-cerebrovascular events (MACCE). Results: Baseline clinical, angiographic, and procedural characteristics were similar between groups, except there was less stent implantation per patient in the observation group (2.13 vs. 2.25 in the control group, P = 0.002). TIMI bleeding (3.3% vs. 4.7%) showed no significant difference between the observation group and control group. During the 30-day follow-up, the rate of MACCE was 0.9% in the observation group and 1.5% in the control group. There was no significant difference in the rates of MACCE, death, myocardial infarction, target vessel revascularization, cerebrovascular event, and angina within 30 days and 1 year after PCI between groups as well as in the subgroup analysis of transfemoral approach. Conclusions: UFH with sequential enoxaparin has similar anticoagulant effect and safety as UFH in PCI of complex coronary artery disease. Key words: Complex Coronary Heart Disease; Enoxaparin; Percutaneous Coronary Intervention; Unfractionated Heparin

Nowadays, blockchain has become one of the most cutting-edge technologies, which has been widely concerned and researched. However, the quantum computing attack seriously threatens the security of blockchain, and related research is still less. Targeting at this issue, in this paper, we present the definition of post-quantum blockchain (PQB) and propose a secure cryptocurrency scheme based on PQB, which can resist quantum computing attacks. First, we propose a signature scheme based on lattice problem. We use lattice basis delegation algorithm to generate secret keys with selecting a random value, and sign message by preimage sampling algorithm. In addition, we design the first-signature and last-signature in our scheme, which are defined as double-signature. It is used to reduce the correlation between the message and the signature. Second, by combining the proposed signature scheme with blockchain, we construct the PQB and propose this cryptocurrency scheme. Its security can be reduced to the lattice short integer solution (SIS) problem. At last, through our analysis, the proposed cryptocurrency scheme is able to resist the quantum computing attack and its signature satisfies correctness and one-more unforgeability under the lattice SIS assumption. Furthermore, compared with previous signature schemes, the sizes of signature and secret keys are relatively shorter than that of others, which can decrease the computational complexity. These make our cryptocurrency scheme more secure and efficient.

Objective To evaluate the safety and cost of early discharge compared with ordinary discharge in patients with diabetes mellitus (DM) following percutaneous coronary intervention (PCI) for acute coronary syndrome. Methods We performed a retrospective analysis of prospectively collected data from 474 patients with DM who were discharged from hospital following PCI at a regional center between 2012 and 2015. Results A total of 192 patients (40.5%) were included in the early discharge group and 282 patients (59.5%) were included in the ordinary group. Mortality and morbidity after PCI were recorded. Kaplan–Meier analysis showed similar prognosis between the two groups at 30 days and at 1 year after discharge. However, hospitalization expenses for the regular discharge group were significantly higher than those of the early discharge group (RMB65,750 vs. RMB50,983). Conclusion Our findings demonstrate that early discharge of patients with DM following PCI for acute coronary syndrome is safe compared with ordinary discharge, and may reduce hospitalization costs.

Digital management of PetroChina Changqing Oilfield Company (PCOC) was a modern management mode for "three-low" oil and gas reservoir economic development. Based on the establishment of digital management "three terminals and five systems", supported by digital technology, we set up one unified platform for production operation management, production operation scheduling, and emergency response and rescue, which provided an efficient collaborative work environment for management decision-making and scientific research. Combined digital technology with management, we promoted a shift in terms of labor organization structure, production organization and production operation mode.

Summary The combination of high pressure and heat on Bacillus cereus spores in food matrix was investigated with the purpose of achieving a predictive model of microbial inactivation. The high-pressure processing (HPP) conditions were fixed at 540 MPa and 71 °C for 16.8 min, which were determined as the optimum conditions considering six-log-cycle reductions of B. cereus spores. The effects of soybean protein, sucrose, soybean oil and pH on the inactivation of B. cereus spores by HPP were evaluated, and a quadratic predictive model for the effects of food ingredients and pH on the reductions of B. cereus spores by HPP was built using response surface methodology. The experimental results showed that soybean protein, sucrose and pH significantly affected the reductions of B. cereus spores. The predictive model is significant, because the level of significance was P

The objective of this investigation was to evaluate the efficacy of high hydrostatic pressure and mild heat against spores of Geobacillus stearothermophilus AS 1.1923 in model food systems. The pressure-processing conditions were fixed at 625.0 MPa and 86C for 14 min, which have been determined as the optimum processing conditions considering six-log-cycle reductions of G. stearothermophilus spores. Based on the results, response surface methodology was performed in the present investigation, the effects from food ingredients, such as soybean protein, soybean oil and sucrose, as well as pH of the food matrix on the inactivation of G. stearothermophilus spores by high pressure and mild heat was explored, and a quadratic predictive model for the effects of food ingredients and pH on the reduction levels of G. stearothermophilus spores by high-pressure processing was built. The predictive model is significant because the level of significance was P

In this study, six Chinese strains of Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) isolated between 1953-1960 were analysed and their molecular characteristics compared to those of the African PG1 and Afade strains, the European C305 and 138/5 strains and the closely related caprine M. mycoides subsp.mycoides large colony type Y-goat strain. PCR amplification of long DNA fragments showed that the six Chinese strains, the PG1 strain and the Y-goat strain, just like Afade, did not have the 8.84 kb deletion characteristic of the European strains C305 and 138/5. In comparison, the lppB gene sequence of the six MmmSC Chinese strains was found to be 99% homologous to that of PG1and Afade, but93% homologous to the Y-goat sequence. The anti-rLppB antiserum reacted with PG1, Y-goat and the six Chinese strains at 67 kDa sites in Western blot, indicating that the lppB gene and its encoding protein exist in the Chinese strains. Multilocus sequence analysis (MLSA) of MmmSC strains from various regions confirmed that the Chinese strains were identical to the African and Australian cluster. This finding was further supported by the outcome of selective primer amplification. Based on these results, it is suggested that CBPP in China may have originated from Australia.

The effects of high pressure (400–600 MPa) and moderate heat (60–80 °C) treatments at various process times (10–20 min) on the reduction of Bacillus cereus As 1.1846 spores, suspended in milk buffer were investigated. In the present work, response surface methodology (RSM) was employed, and a quadratic equation of high hydrostatic pressure inactivation was built with RSM. By analyzing response surface plots and corresponding contour plots and by solving the quadratic equation, experimental values were shown to be significantly in agreement with predicted values, since the adjusted determination coefficient ( R Adj 2 ) was 0.9752 and the level of significance was P

Our published results and our studies for optimization of process conditions to inactivate Bacillus subtilis by high hydrostatic pressure and mild heat using response surface methodology indicated that the optimum process parameters for a six-log-cycle reduction of B. subtilis were obtained as temperature, 46 °C; pressure, 479 MPa; and pressure holding time, 14 min. Based on the results, response surface methodology (RSM) was employed in the present investigation, the effects of food constituents like soybean protein, soybean oil, sucrose, and pH of food matrix on the B. subtilis reduction during high pressure and moderate heat was studied, and a quadratic polynomial predictive model for the effects of food constituents and pH of food matrix on B. subtilis reduction during high pressure and moderate heat was built with RSM accurately. The experimental results showed that the efficiencies of B. subtilis reduction in milk buffer and food matrix designed in the present work, under the condition of high pressure treatment process parameters described above, had some differences. The soybean protein ( P P P = 00006) significantly affected reduction of B. subtilis . The effect of soybean oil on reduction of B. subtilis was not significant ( P = 0.8363). The adequacy of the predictive model equation for predicting B. subtilis reduction in food matrix by high pressure and moderate heat was verified effectively using experimental test date that was not used in the development of the model.

Our formerly published experimental results of studies on inactivation of Bacillus subtilis As 1.173 spores by high hydrostatic pressure and heat using design of experiments showed that the optimum process parameters for a six log-cycle reduction of spores of B. subtilis were obtained as: pressure, 576.0 MPa; temperature, 87 °C; and pressure holding time, 13 min. Based on the results, response surface methodology (RSM) was performed in the present investigation, to determine effects of food composition (soybean protein, bean oil, sucrose) and pH on the inactivation of B. subtilis As 1.173 spores by high pressure and mild heat were studied. A quadratic predictive model was built for the effects of food compositions and pH on the reduction levels of B. subtilis spores by HPP using RSM. The experimental results showed that the reduction of B. subtilis spores in buffer and foods differed depending on the HPP treatment process parameters. The soybean protein ( P = 0.0005), sucrose ( P = 0.0003), and pH ( P = 0.0080) significantly influenced the reduction of B. subtilis spores, and the effect of bean oil on the reduction of B. subtilis spores was only slightly significant ( P = 0.0810). Moreover, the adequacy of the model equation for predicting the reduction of B. subtilis spores was verified effectively using experimental test data that was not used in the development of the model.

The combination of high hydrostatic pressure with mild temperature was explored to achieve a predictive model of microbial inactivation in food matrix processing. The pressure processing conditions were fixed at 448 MPa for 11 min at the treatment temperature of 41 degrees C, which have been determined as the optimum processing conditions considering six log-cycle reductions of Listeria monocytogenes. Based on the results, response surface methodology (RSM) was performed in the present work, the influence of food components like soybean protein (0-5.00%), sucrose (0.25-13.25%), bean oil (0-10.00%), and pH (4-10) of the food matrix on survival of L. monocytogenes by high pressure and mild heat was studied, and a quadratic predictive model for the influence of food components and pH of food matrix on L. monocytogenes reduction by high pressure and mild heat was built with RSM accurately. The experimental results showed that the efficiency of L. monocytogenes reduction in milk buffer and food matrix designed in the present work, under the HPP treatment process parameters described above, were different. The soybean protein (P=0.0086), sucrose (P

The results of fractional factorial experiments showed that the significant external factors affecting high-pressure processing (HPP) inactivation were pressure, temperature and pressure holding time. Based on these results, response surface methodology (RSM) was employed in the present work and a quadratic equation for HPP inactivation was built. By analyzing the response surface plots and their corresponding contour plots as well as solving the quadratic equation, the experimental values were shown to be significantly in good agreement with predicted values since the adjusted determination coefficient ( R Adj 2 ) was 0.9384. The optimum process parameters for six log-cycles reduction of Bacillus subtilis spores were obtained as: temperature, 87 °C; pressure, 576.0 MPa and pressure holding time, 13 min. The adequacy of the model equation for predicting the optimum response values was verified effectively by the validation data.

Previous unpublished experimental results of fractional factorial experiments showed that significant external factors affecting high-pressure processing (HPP) inactivation were pressure, temperature, and pressure holding time. Based on these results, response surface methodology (RSM) was employed in the present work, and a quadratic equation for HPP inactivation was built with RSM. By analyzing response surface plots and their corresponding contour plots and by solving the quadratic equation, experimental values were shown to be significantly in good agreement with predicted values, since the adjusted determination coefficient (RAdj2) was 0.9812 and the level of significance was P0.0001. Optimum process parameters for a 6-log cycle reduction of Listeria monocytogenes were obtained: pressure, 448.0 MPa; temperature, 41 degrees C; and pressure holding time, 11 min. The adequacy of the model equation in predicting optimum response values was verified effectively by validation data.

The previous experimental results for effect of high pressure and mild heat on Staphylococcus aureus in milk using response surface methodology showed that the optimum process parameters for six log-cycles reduction of S. aureus were obtained as: temperature, 34.4 °C; pressure, 329.8 MPa; pressure holding time, 15.5 min. Based on the results, response surface methodology (RSM) was employed in the present work, the effect of food main constituents like soybean protein, sucrose, bean oil and pH of the food matrix on the S. aureus inactivation by high pressure and mild heat was studied, and a quadratic predictive model for the effect of food constituents and pH of food matrix on S. aureus reduction by high pressure and mild heat was built with RSM accurately. The experimental results showed that the efficiencies of S. aureus reduction in milk and food matrix designed in the present work under the HHP treatment process parameters described above had some differences. The soybean protein (P = 0.0082), sucrose (P

The results of fractional factorial experiments showed that the significant external factors affecting high pressure processing (HPP) inactivation were pressure, temperature and pressure holding time. Based on these results, response surface methodology (RSM) was employed in the present work and a quadratic equation for HPP inactivation was built. By analyzing the response surface plots and their corresponding contour plots as well as solving the quadratic equation, the experimental values were shown to be significantly in good agreement with predicted values since the adjusted determination coefficient ( R Adj 2 ) was 0.9800. The optimum process parameters for six log-cycles reduction of Bacillus subtillis were obtained as: temperature, 46 °C; pressure, 479.0 MPa and pressure holding time, 14 min. The adequacy of the model equation for predicting the optimum response values was verified effectively by the validation data.

The results of fractional factorial experiments showed that the significant external factors affecting high pressure processing (HPP) inactivation were pressure, temperature and pressure holding time. Based on these results, response surface methodology (RSM) was employed in the present work and a second order quadratic equation for HPP inactivation was built. By analyzing the response surface plots and their corresponding contour plots as well as solving the quadratic equation, the experimental values were shown to be in good agreement with predicted values since the correlation coefficient was 0.9891. The optimum process parameters for six log-cycles reduction of Staphylococcus aureus were obtained as: temperature, 34.4 °C; pressure, 329.8 MPa and pressure holding time, 15.5 min. The adequacy of the model equation for predicting the optimum response values was verified effectively by the validation data.

To observe the clinical and coronary angiographic features of young women (≤ 44 years) with acute myocardial infarction (AMI).Clinic and coronary angiographic features were compared among 57 young women with AMI and 60 non-CHD young women, 78 young men with AMI and 80 elderly women with AMI were included, all patients were admitted to hospital from June 2003 to December 2010 and underwent coronary angiography. Body mass index (BMI), smoking history, familial history of early coronary artery disease, essential hypertension, type 2 diabetes mellitus, hyperlipidemia, serum uric acid (UA), menopause, lipids, total bilirubin (TBil), direct bilirubin (DBil), hemoglobin (Hb), red cell distribution width (RDW), fasting blood glucose (FBG) were measured.(1) Prevalence of essential hypertension, hypertriglyceridemia and levels of fasting blood glucose, triglyceride, serum uric acid were significantly higher while high density lipoprotein cholesterol, and hemoglobin were significantly lower in young women with AMI group than in age-matched non-CHD control group (all P0.05). (2) Logistic regression analysis showed that essential hypertension (OR = 23.187), lower hemoglobin level (OR = 1.010) and uric acid (OR = 1.040) were independent risk factors for young women with AMI (all P0.05). (3) Coronary angiography revealed normal coronary finding in 6 cases, single-vessel disease in 35 cases (26 cases with left anterior descending artery disease) and two-vessel disease in 12 patients. The ratio of single vessel disease involved left anterior descending artery in young women was higher than that of young men and old women with AMI (all P0.05).Essential hypertension, lower hemoglobin and uric acid are risk factors of young women with AMI. Single vessel coronary disease is the most common coronary angiographic feature of young women with AMI.

Left ventricular hypertrophy associated with the use of tacrolimus is a rare complication of solid organ transplantation in adult recipients. We present a cardiac transplant recipient who developed severe concentric left ventricular hypertrophy with congestive heart failure related to myocardial hypertrophy on tacrolimus. Hypertrophy improved when the drug was discontinued and replaced with sirolimus.

Recombinant baculovirus containing sigmaC gene of Avian reovirus was constructed using Bac-To-Bac Baculovirus expression system, and recombinant sigmaC protein was expressed by infecting the sf9 cell with recombinant baculovirus. Firstly, sigmaC gene of Avian reovirus was cloned and inserted into donor plasmid pFastBacHTA to obtain recombinant donor plasmid pFsigmaC. Plasmid pFsigmaC was transformed into E. coli DH10Bac for integration into bacmid vector and the recombinant bacmid plasmid BacmidsigmaC was obtained. Recombinant baculovirus rBacsigmaC was obtained by transfection of the sf9 cells with BacmidsigmaC. Western blot and indirect immunofluorescence assay (IFA) were carried and the results showed that the recombinant sigmaC protein with 37 kDa molecular weight was expressed successfully.

Endothelial lipase, which is a newly identified member of the lipase family, plays an important role in high-density lipoprotein metabolism, which catalyzes the hydrolysis of high-density lipoprotein phospholipids and facilitates the clearance of high-density lipoprotein from the circulation. In addition, inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta), upregulate endothelial lipase expression, and endothelial lipase also affects the expression of cytokines, which in turn play an important role in atherogenesis. Endothelial lipase expression has been associated with macrophages within human atherosclerotic lesions. However, an important challenge is to determine how endothelial lipase alters the progression of atherosclerosis. Although few data are available from human studies, it seems that plasma endothelial lipase levels in individuals with atherosclerosis might be higher than that measured in healthy individuals. Therefore, we believe that endothelial lipase might be a promising marker for atherosclerosis in clinical settings in the future.

The complete genomic sequence of very virulent infectious bursal disease virus (vvIBDV) Gx strain was determined, including the sequences of segment A, encoding the precursor polyprotein, and segment B, encoding the viral RNA polymerase (VP1) and 5'- and 3'-untranslating regions. Alignment of segment A of Gx with the sequences of 12 other vvIBDV strains showed 97.5% to 99.0% amino acid identity, whereas alignment of segment B of Gx with nine other vvIBDV strains revealed high sequence divergence, ranging from 10.3% to 11%. Phylogenetic analysis of segments A and B showed that they were in different branches, indicating that the reassortment occurred in this strain and that segment A and segment B derived from different pathotype strains. The mutant spectrum analysis of quasispecies virus demonstrated that the mean minimum mutation frequency in VP1 was 8.78-fold higher than in the polyprotein. The most frequent mutations were in the first 1986 nucleotides (nonsynonymous mutations) and the last 660 nucleotides (synonymous mutations), indicating that the 219 amino acid residues in the C-terminal of the VP1 form a functional region.

Infectious bursal disease virus(IBD) causes infectious bursal disease (IBD), which infects bursal of chicken and can evoke immune suppression. This study identified the antigenic epitopes of four McAbs to IBDV VP3(HRB-3F, HRB-7B, HRB-7C and HRB-10E)with pepscan. A set of 17 partially overlapping or consecutive peptides (P1-P17) spanning VP3 were expressed for epitope screening by pepscan. Finally, two antigenic epitopes, 109-119aa and 177-190aa of IBDV VP3, were identified by Western blot and ELISA. The peptides on epitopes could react with IBDV, and they had better immunnogenicity. The sequences of epitopes were compared with that of several other IBDV strains in the same region, and was found they were totally homologous. This study showed the two epitopes were novel conserved linear B cell epitopes on the VP3 of IBDV. This study provides basis for the development of immunity-based prophylactic, therapeutic and diagnostic measures for control of IBD and further for structural and functional analysis of IBDV.

In the present work, we studied the combined effects of pressure (300.0-700.0 MPa), temperature (30-70 degrees C) and the presence of nisin (0-333 IU/ml) on the inactivation of Clostridium botulinum 33A spores at various pressure holding times (7.5-17.5 min). Moreover, response surface methodology (RSM) was employed and a quadratic equation for HPP and nisin-induced inactivation was built with RSM. By analyzing the response surface plots and their corresponding contour plots as well as solving the quadratic equation, the experimental values were shown to be significantly in good agreement with predicted values because the adjusted determination coefficient (R(Adj)(2)) was 0.9261 and the level of significance was P

Infectious bursal disease virus (IBDV), the causative agent of a highly contagious disease in chickens, carries a small nonstructural protein (NS). In this study, vvIBDV Gx-VP5 genes were cloned into plasmid pET30a( + ) and expressed in E. coli with IPTG inducing. BALB/c mice were immunized with the purified recombinant fusion protein. SP2/0 myeloma cells and spleen cells of BALB/c mice were fused by PEG(MW1500), three hybridoma cell lines were examined by indirect ELISA and clone for three times by limited dilution, and were named as 4B4, 6D12, 3E8. The subtype of the monoclonal antibodies were IgG1 with a subtype identified ELISA kit, and light chains were kappa. The ascites titers of monoclonal antibodies were 5 x 10(4), 3.5 x 10(4), 3 x 10(4) by indirect ELISA, respectively. Indirect ELISA and Western blot results showed that the monoclonal antibodies only acted with VP5 protein, IF analysis indicated that three monoclonal antibodies acted with IBDV Gt. There were specific fluorescence in detected Vero E6 cells which transient expressed VP5 protein by IFA. Therefore, monoclonal antibodies specific to IBDV VP5 proteins are specific method for detected VP5 proteins, and base on establish stabilize expressed VP5 protein Vero cell lines to research IBDV VP5 protein function.

Chicken embryo fibroblast (CEF) is a primary cellular material to research the infectious bursal disease virus (IBDV). Constructing the cDNA expression library of CEF is the foundation to research cell tropism and find cell receptors of IBDV from CEF. In order to achieve that purpose, a high-quality cDNA expression library of CEF was constructed by Gateway technology, which could avoid using the restriction enzyme for cloning to solve technical limitation of roution method. The mRNA was extracted from chicken embryonic fibroblast. Moreover, single-strand cDNA and double-strand cDNA were synthesized by using biotin-conjugated Oligo (dT) primer in turn. The double-strand cDNA was ligated Adapter and then purified by the cDNA Size Fractionation Columns. After BP recombination reaction, a cDNA entry library was constructed with a titer of 1 x 10(6) cfu/mL, total clones of 1.2 x 10(7) cfu and an average insertion size of about 2243 bp. After LR recombination reaction, the cDNA entry library was transformed into expression library which took on a titer of 5 x 10(5) cfu/mL, total clones of 5.5 x 10(6) cfu and an average insertion size of about 2411bp. The results indicate that the constructed cDNA expression library performs a remarkable high value in both recombination rate and library coverage. As a result, the cDNA expression library, with its good quality, may facilitate to identify the receptors associated with the resistance against IBDV in chicken embryonic fibroblast and to cast new light on the mechanism of cellular tropism. Moreover, it may also provide data of chicken embryonic fibroblast in transcription level and may be helpful to study its biological functions.

The coding region of VP2 gene from Chicken Infectious Anemia was amplified from genome extracted from chicken liver tissue by PCR. PCR product was double digested with restriction enzymes BamH I and Sal I and cloned into pET-28a digested with BamH I and Sal I. Subsequently, the recombinant plasmid pET-28-VP2 was extracted and double digested with restriction enzymes BamH I and Sal I. After confirming its rightness by PCR and analysis of restriction endonucleases, the recombinant plasmid pET-28-VP2 was transformed into E. coli BL21 (DE3) strain. The culture was induced by 1 mmol/L IPTG at 37 degrees C for three hours and analyzed with SDS-PAGE. The result shows that gene encoding VP2 of CIAV was expressed successfully in E. coli and the fusion protein existed in supernatant, which was about 31kDa and showed specific immunoreactivity with anti-CIAV sera in Western blot. The fusion protein was purified by Ni2+ -affinity chromatography and quantitated by Bradford method. Then BALb/c mice were immunized with purified protein emulsified with Freund's complete adjuvant on day 0 and boosted twice on day 14 and 28 with the same dose of antigens emulsified with Freund's incomplete adjuvant, respectively. The serum isolated were examined by an enzyme-linked immunosorbant assay (ELISA) using the purified VP2 and CIAV as coating antigens and the serum could react with target protein and CIAV in ELISA detection test.