Interferon-tau (IFN-tau) is a newly discovered IFN of type I. It was originally found for its role as a pregnancy recognition hormone in ruminant animals such as sheep and cows. Like the other type I IFNs, IFN-tau have the same biological activities including antiviral, antiproliferative and immunomodulatory effects. In order to clearly identify the function of IFN-tau, the coding sequence of IFN-tau was amplified by PCR from IFN-tau cDNA, this fragment digested by EcoR I and BamH I and was inserted into pBV220 to form the recombinant plasmid pBV220/IFN-tau which was then transformed into E.coli BL21. It was found that pBV220/IFN-tau was highly expressed as inclusion body in BL21. Next, the expressed protein was purified on S-100 High Resolution and the purified product was confirmed by amino acid sequence analysis. Moreover, the standard antiviral activity test indicated that the activity of IFN-tau was about 2.09?106IU/ml.

Objective To investigate the effects of ketamine on the expression of NMDA receptor-1(NRⅠ)in a rat model of focal cerebral ischemia and the possible mechanism of the neuroprotection.Methods Forty healthy male SD rats weighing 250-290g were randomly divided into 2 group(n=20 each):groupⅠketamine and groupⅡpentobarbital.The aminals were anesthetized with intraperitoneal ketamine 60 mg?kg~(-1) in groupⅠor pentobarbital 40 mg?kg~(-1) in groupⅡ.Focal cerebral ischemia was produced by permanent middle cerebral artery occludion(MCAO).The animals were killed at 24 h and 72 h of MCAO and their brains removed for determination of infarct size,the number of living neurons in the penumbra and the expression of NRⅠprotein(immuno- histochemistry).Results The infarct size was significantly smaller;the number of living neurons in penumbra significantly larger and NRⅠexpression significantly down-regulated in ketamine group than in pentobarbital group.Conclusion Ketamine can protect the brain against ischemia through downregulation of NMDA receptor-1.

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.

Objective To summarize the clinical characteristics,diagnosis and treatment of malignant tumor after renal transplantation.Methods The clinical data of 2106 renal transplants in 1945 patients undertaken in our hospital from September 1978 to December 2009 was retrospectively studied.Results Of these 1945 patients, 39 cases were diagnosed as having malignant tumor (incidence: 2.0 ％).The interval between transplantation and clinical diagnosis ranged from 8 to 124 months with a median of 57.0 months.Among the 39 cases of malignancy, there were 22 urinary system carcinomas, 8 digestive system carcinomas,2 lung cancers,2 breast cancers,2 lymphomas,1 dura small cell carcinoma,1 pleura poorly differentiated carcinoma and 1 metastatic carcinoma of liver with unknown primary tumor. Surgery was conducted in 28 patients,of which 16 were survived but the other 12 patients died of metastasis ranged from 3 months to 96 months (median,33 months) after operation.11 cases without operation died within from 3 d to 36 months (median,5 months) after diagnosis.Conclusions The incidence of malignant tumors in renal transplant recipients increased markedly.The most common type of the malignant tumors is urinary system carcinoma.The key measure of success in treating malignancy after renal transplantation is early diagnosis and surgical resection.

HIV p24 core protein can induce both cellular and neutralizing antibody responses.HIV-1 CA-virus-like particles(VLPs)vaccines provide a promising approach for the development of an effective vaccination strategy against HIV infection.Rhizosecreion of the recombinant proteins provides a new manufacturing platform that can simplify the extraction and purification procedure.Lycium barbarum L.was transformed by Agrobacterium tumefaciens EHA105 harboring the plant expression vector pCAMBIA1305.2-MA4-CA with a GRP signal peptide and MA4-CA fusion gene.Transgenic hairy roots were induced and cultivated in hydroponic culture.Western blotting indicated that the recombinant CA proteins were present in two forms,a glycosylated monomer(37 kDa)and a dimer(50 kDa)in the roots and hydroponic medium.It appeared from the present immunohistochemical data that the recombinant CA proteins fused with GRP signal peptide were confined to the cytoplasm,cell wall and intercellular space,indicating targeting into the secretory pathway.It demonstrated for the first time the rhizosecretion of HIV-1 recombinant capsid protein in Lycium barbarum L.hairy roots,and may offer a novel method for expressing HIV-1 CA-VLPs vaccines in plants.

Objective:To compare the therapeutic efficacy and safety of Hyper-CVAD/MA regimen and CHOP/CHOP-like regimen in the treatment of peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS). Methods:The 78 primary PTCL-NOS patients who were initially diagnosed and treated in Tianjin Medical University Cancer Institute and Hospital and Tianjin Union Medical Center from June 2004 to June 2012 were retrospectively analyzed. The patients were then divided into two groups:Hyper-CVAD/MA group (n=21) and CHOP/CHOP-like group (n=57). Curative efficacies and toxicities were analyzed by Chi-square test, and survival was estimated by Ka-plan-Meier method. Results: In the Hyper-CVAD/MA group, complete response (CR) was 42.9%, overall response rate (ORR) was 85.7%, median progression-free survival (PFS) was 20 months, and the three-year overall survival (OS) was 56.9%. In the CHOP/CHOP-like group, the CR, ORR, and three-year OS were 28.1%, 59.6%, and 49.6%, respectively, and the median PFS was 13 months. Compara-tive analysis showed that the ORR and three-year OS were statistically significant (P<0.05), but the relapse rates (57.1%versus 77.2%) and three-year OS were similar (P>0.05). The incidence rates ofⅢ/Ⅳneutrocytopenia and thrombocytopenia in Hyper-CVAD/MA group (66.7%and 61.9%, respectively) were significantly higher than those of the CHOP/CHOP-like group (22.8%and 14.0%, respec-tively) (P<0.05). Conclusion:Hyper-CVAD/MA regimen can achieve satisfactory efficacy in parents with PTCL-NOS, and toxicity can be controlled with granulocyte colony stimulating factor (G-CSF).

OBJECTIVETo screen human stem cell factor (hSCF) mimetic peptides in vitro with a phage-display random peptide library.

METHODSPhage clones with high hSCF receptor (rc-kit/Ig 1-3)-binding activity was screened from phage-displayed random hepta/dodecapeptide library by phage enzyme-linked immunosorbent assay (ELISA). Phage single DNA was extracted and sequenced. Four kinds of peptide with higher c-Kit/Ig 1-3 binding activity were chosen for synthesis and characterized by using cell proliferation assay with 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) method in UT-7 cells.

RESULTSEleven Ph.D.-C7C clones and eight Ph.D-12 phage clones with high hSCF receptor-binding activity were selected from phage-displayed random hepta/dodecapeptide library, respectively. Sequence analysis showed there were no homologous sequence between hSCF and these screened mimetic peptides except one homologous sequence DPSPHTH found in heptapeptide library. All these four synthesized peptides (CE3, CE16, LE4, and LE20), particularly CE16 and LE20, stimulated UT-7 cell proliferation.

CONCLUSIONFour hSCF mimetic peptides were successfully isolated from phage-displayed random peptide library..

Humans ; Peptide Library ; Peptides ; genetics ; isolation & purification ; Stem Cell Factor ; genetics ; isolation & purification

Objective To investigate the distribution and antimicrobial resistance profile of the common pathogens isolated during the period from 2009 to 2015.Methods All the bacterial strains isolated from pediatric inpatients in Beijing Children's Hospital during the period from 2009 to 2015 were analyzed. Antimicrobial susceptibility was determined by disk diffusion method and Phoenix 100 Automated Microbiology System. Results were analyzed according to the guidelines of CLSI (2014) using WHONET 5.6 software.Results The total strains were 26630. The most common gram-positive isolates were Streptococcus pneumoniae,Staphylococcusaureusand coagulase-negative Staphylococcus (CNS), while the most frequently isolated gram-negative microorganisms were Klebsiella spp.,Pseudomonas aeruginosa and Escherichia coli. The prevalence of S. pneumoniae was up to 25.7 % (4101/15973) in all respiratory tract specimens. About 50.2 % of the S. pneumoniae isolates were not susceptible to penicillin. The prevalence of methicillin-resistant strains was 20.6 % in S. aureus (MRSA) and 87.8 % in coagulase negative Staphylococcus (MRCNS) on average. The prevalence of MRSA increased from 11.1 % in 2009 to 29.8 % in 2015. No S. pneumoniae or staphylococcal strains were found resistant to vancomycin or linezolid. The Enterococcus strains were still highly susceptible to vancomycin and linezolid. Overall 0.3 % of the Enterococcus faecium isolates were resistant to vancomycin. The extended-spectrum beta-lactamases (ESBLs) producing strains accounted for 71.4 % -78.1 % of E. coli and 65.1 % - 76.9 % of K. pneumoniae isolates. The carbapenem-resistant E. coli and K. pneumoniae were reported for the first time in 2010, but in 2014, the strains resistant to carbapenems had increased to more than 7 % in E. coli, and higher than 20 % in K. pneumoniae. In 2015, up to 27.7 % and 25.7 % of P. aeruginosa isolates were resistant to imipenem and meropenem, respectively, and 59.9 % of the A. baumannii isolates were resistant to imipenem and meropenem. Beta-lactamase was positive in 46.3 % of the H. influenzae isolates. Conclusions MRSA and the carbapenem-resistant strains of E. coli,K. pneumoniae and A. baumannii are still on the rise in pediatric inpatients, which poses a serious threat to clinical practice and implies the importance of strengthening infection control.

OBJECTIVETo explore the clinical value of determination of ATP levels in CD4(+) cells of patients with cytomegaloviral pneumonia after kidney transplantation.

METHODSTwenty-eight patients with cytomegaloviral pneumonia following kidney transplantation and 30 healthy volunteers were enrolled in this study. ATP-bioluminescence assay (ATP-CVA) was used to assess the immune response of CD4(+) cells to phytohemagglutinin (PHA) stimulation in the normal volunteers and the recipients (before and at 1, 2, and 4 weeks after renal transplantation, before and at 2 and 4 week after the treatment).

RESULTSATP concentration in CD4(+) cells of the recipients was 402-/+58 ng/ml before the operation, significantly lower than that in normal volunteers (458-/+196 ng/ml, P<0.05), and reached the lowest level in the first week after operation especially in the recipients with antibody-inducing therapy; ATP level increased slowly since week 2 post-operation, but still remained significantly lower than the preoperative by the fourth week (266-/+87 ng/ml, P<0.05), especially in the recipients receiving antibody-inducing therapy. In the event of cytomegaloviral pneumonia, ATP level underwent a mild reduction to 152-/+78 ng/ml in comparison with the postoperative level at the first week (P>0.05), and was significantly lower than preoperative level (P<0.01); the decrease was especially obvious during the exacerbation of the condition. ATP level then increased slowly after effective treatment, but was still lower than the preoperative level at 4 weeks after the operation (336-/+92 ng/ml, P<0.05).

CONCLUSIONThe determination of ATP level in CD4(+) cells allows more accurate assessment of the cellular immunity in the renal transplant recipients with cytomegaloviral pneumonia to help in the clinical treatment of the patients.

Adenosine Triphosphate ; blood ; Adult ; Aged ; CD4-Positive T-Lymphocytes ; metabolism ; Case-Control Studies ; Cytomegalovirus Infections ; immunology ; Female ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Pneumonia, Viral ; immunology ; metabolism ; virology ; Postoperative Complications ; immunology ; metabolism ; Young Adult

OBJECTIVETo determine the lung expression of tissue factor (TF) mRNA in hepatopulmonary syndrome (HPS) using a rat model system and to investigate the potential significance of its differential expression.

METHODSForty male Sprague-Dawley rats were used to establish models of cirrhosis (n = 20) and HPS (n = 20). Blood gas analysis was used to investigate the effects of each model on pulmonary function. Effects on the expression of TF mRNA in lung were determined by qRT-PCR and on lung pathology by histological analysis.

RESULTSThe HPS rats showed significantly lower PaO2 than the cirrhosis rats (58.20 +/- 3.19 mmHg vs. 85.00 +/- 2.53 mmHg, P less than 0.05) but significantly higher TF mRNA expression in lung (0.77 +/- 0.22 vs. 0.33 +/- 0.14, P less than 0.05). TF mRNA expression was negatively correlated with the value of PaO2 (r = -0.565, P less than 0.05). The lungs of the cirrhosis rats showed widened alveolar intervals, diversified sizes of alveolar spaces, reduced lung capacity, inflammatory cell infiltration, and hyperemia in the pulmonary vessels. The lungs of the HPS rats showed all of the same changes but also with accumulated macrophages and micro-thrombosis in the pulmonary vessels. Among the HPS rats, those with micro-thrombosis in pulmonary vessels showed a greater increase in TF mRNA expression than those without (0.68 +/- 0.17 vs. 0.40 +/- 0.12, P less than 0.05).

CONCLUSIONThe expression of TF mRNA in lung of hepatopulmonary syndrome model rats was elevated and might increase the incidence of thromboembolism in the lung.

Animals ; Disease Models, Animal ; Hepatopulmonary Syndrome ; genetics ; metabolism ; Lung ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thromboplastin ; genetics ; metabolism