Objective To investigated the expression of intracellular adhesion molecule-1 (ICAM-1)in mouse to rat cardiac xenograft and the effects of leflunomide and cyclosporine.Methods NIH mice and Wistar rats served as donors and recipients respectively.Mouse to rat heterotopic heart transplantation was performed(in the neck).The experiments were divided into 4 groups:CsA group,leflunomide(Lef)group,CsA+Lef group,control group.It was the time of rejection that no pulsation could be detected in the transplanted heart.The expression of ICAM-1 protein in cardiac xenograft tissue was detected by immunohistochemistry and Western blot in each group.Results The survival time of cardiac xenograft in control group,CsA group,Lef group and CsA+Lef group was (2.17?0.41),(2.50?1.05),(4.17?1.33)and(6.50?2.56)days respectively.The expression of ICAM-1 protein(IOD)in cardiac xenograft tissue in control group,CsA group,Lef group and CsA+ Lef group was 155.40?5.33,150.73?5.13,104.65?6.15 and 29.24?2.76 respectively.Conclu- sion The expression of ICAM-1 protein was strong in mouse to rat cardiac xenograft.The combined use of Lef and CsA can significantly suppress the expression of ICAM-1 protein in the cardiac xenograft.

Based on site-specific transposition of an expression cassette into a baculovirus shuttle vector (Bacmid) which propagated in Escherichia coli, the Bac-to-Bac System provides a rapid and efficient method to generate recombinant baculoviruses and is widely used for high level expression of heterologous proteins. And the efficiency of recombinant baculovirus infecting cells plays an important role on the protein expression. In this study, we introduced an EGFP expression cassette driven by polyhedrin promoter into the p74 locus of Bacmid by homologous recombination. The target Bacmid-egfp was then transformed into E. coli DH10B containing the transposition helper plasmid to gain a new transposition receipt strain E. coli DH10Bac-egfp. Because of the intact attTn7 sites and lacZ', target gene cloned in a pFastBac vector can be transposed into the Bacmid-egfp shutter vector to construct recombinant baculovirus, which would allow the tracing of the target protein expression and the recombinant Bacmid transfection or recombinant baculoviral infection under fluorescence microscopes. Recombinant virus Bac-egfp-DsRed was constructed by transposing DsRed into the Bacmid-egfp in E. coliDHl0Bac-egfp, and the Sf9 cells infected with the recombinant virus expressed DsRed and EGFP efficiently. Another protein IL-6 fused with 6 x his tag was expressed and purified sucessfully from Sf9 cells infected with recombinant virus Bac-egfp-6 x his-IL6 constructed by the improved Bac-to-Bac system.
Animals ; Baculoviridae ; genetics ; Green Fluorescent Proteins ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Spodoptera

In this study, a 15-mer phage display peptide library was employed to pan against human rotavirus immobilized on solid phase. 4 different peptides were selected and could bind with rotavirus particles specifically. Plaque reduction neutralization test and MTT analysis results indicated that 3 of the peptides can inhibit rotavirus infecting in vitro. A peptide which sequence is QSNPIHIITNTRNHP showed the best efficiency--93% neutralization infectivity. Two other peptides, A and B, showed 40% and 50% neutralization infectivity respectively. Amino sequence analysis results indicate the 3 peptides containing 2 conserved motifs: SNPIHII and NIP. No putative trypsin hydrolysis site was found in C peptide, however, 4 and 3 potential sites were found in A and B peptides respectively. Using trypsin inhibitor, both A and B peptides showed the similar antiviral effect as that of C peptide. It suggests that the intactness of the 2 conserved motifs play an important role in counteracting virus infection. According to the results of this study, peptide C is hopeful to be exploited as an antiviral peptide drug.
Amino Acid Sequence ; Animals ; Antiviral Agents ; isolation & purification ; pharmacology ; Cell Line ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drug Evaluation, Preclinical ; Humans ; Molecular Sequence Data ; Neutralization Tests ; Peptide Library ; Peptides ; chemistry ; immunology ; pharmacology ; Protein Binding ; Rotavirus ; drug effects ; growth & development ; immunology ; Sequence Analysis, Protein ; Viral Plaque Assay

OBJECTIVETo analysis the efficacy and safety of lamivudine (made in China) therapy for 52 weeks in adolescent patients with chronic hepatitis B (CHB).

METHODSOne hundred and five teenage CHB patients were treated with lamivudine 100 mg once daily for 52 weeks. Patients with elevated ALT at baseline were in group 1 and those with normal ALT were in group 2. The changes of HBV DNA, HBV seromarkers and ALT at the end of 12, 24 and 52 weeks after lamivudine therapy were compared with those at baseline. Adverse events were recorded and evaluated.

RESULTSAt the end of 52 weeks of lamivudine therapy, HBV DNA-ve, HBeAg loss and anti-HBe seroconversion were observed in 92.0%, 24.4% and 22.0% in group 1 patients and 76.1%, 14.2% and 14.2% in group 2 patients respectively. No significant differences were found between two groups. At 12, 24 and 52 weeks, normalization rates of ALT were 59.0%, 66.7% and 76.0%, normal ALT with undetectable HBV DNA were 44.9%, 64.1% and 70.7% at the same time. During 52 weeks lamivudine treatment 26 mild adverse events were observed in 18 patients.

CONCLUSIONLamivudine can inhibit HBV replication rapidly and normalize ALT in majority adolescent CHB patients. HBeAg loss or seroconversion of anti-HBe was observed in some of these patients. All patients in this study were safety and well tolerated.

Adolescent ; Alanine Transaminase ; blood ; Antiviral Agents ; adverse effects ; therapeutic use ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B e Antigens ; blood ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Lamivudine ; adverse effects ; therapeutic use ; Male ; Mutation ; genetics ; Reverse Transcriptase Inhibitors ; adverse effects ; therapeutic use ; Safety ; Treatment Outcome

OBJECTIVETo study eleven organophosphorus insecticides residuals in four kinds of Chinese crude drugs.

METHODThe organophosphorus insecticides were extracted with dichloromethane and cleaned-up with a mixture of Celite 545-activated carbon. The extracts were analyzed by gas chromatography equipped with a flame photometric detector (FPD).

RESULTAnalysis of fortified Chinese crude drug showed that the average recoveries ranged from 77.5% -112.3% at three different levels, the RSDs were below 10% (n = 4). Trace organophosphorous pesticide residues were found in samples of Rhizoma Atractylodis Macrocephalae and Flos Chrysanthemi.

CONCLUSIONA method was established for determination multi-residues in Rhizma Atractylodis Macrocephalae, Radix Curcumae, Bulbus Fritillariae Thunbergii and Flos Chrysanthemi. It provides a method for the risk assessment of organophosphorous pesticide in Chinese crude drugs.

Atractylodes ; chemistry ; Chromatography, Gas ; Chrysanthemum ; chemistry ; Curcuma ; chemistry ; Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Fritillaria ; chemistry ; Organophosphorus Compounds ; analysis ; Pesticide Residues ; analysis ; Plants, Medicinal ; chemistry

OBJECTIVETo introduce and evaluate the clinical effects of percutaneous reduction and Kirschner pin fixation for the treatment of intraarticular fractures of the calcaneus in children.

METHODSFrom March 2001 to February 2009,12 patients with intraarticular calcaneal fractures were treated by percutaneous reduction and Kirschner pin fixation (13 feet). There were 8 males and 4 females,ranging in age from 3 to 14,with an average of 8.7 years. According to Essex-Lopresti classification, among 5 feet were tongue fractures and 8 feet were compressed fractures. According to Sanders classification, 9 feet were type II and 4 feet were type III. The Biihler angle and Gissane angle of the calcaneus were obtained before and after operation. All patients were evaluated according to Maryland Foot Score.

RESULTSAll the patients were followed up for 16-71 months (means 35.9 months),and all the incisions were healed without complications and infection. The preoperative X-ray film showed that Böhler angle was (19.7+/-5.3) degrees, Gissane angle was (137.3+/-7.5) degrees. The postoperative X-ray film demonstrated that Böhler angle was (32.6+/-3.7) degrees, Gissane angle was (125.4+/-2.9) degrees. There was a significant difference between preoperative and postoperative (P<0.01). The average Maryland score was 96.3+/-2.4 (range, 92 to 100 points).

CONCLUSIONPercutaneous reduction and Kirschner pin fixation is an effective minimally invasive way to treat intraarticular fractures of the calcaneus in children, it has many advantages such as minimal invasion, reliable fixation and satisfactory effects.

Adolescent ; Bone Nails ; Calcaneus ; injuries ; surgery ; Child ; Child, Preschool ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Intra-Articular Fractures ; surgery ; Male

OBJECTIVETo study the clinical effects of AO distal humerus plate (DHP) for the treatment of distal humeral fractures in elderly osteoporotic patients.

METHODSFrom September 2008 to January 2010, 18 elderly osteoporotic patients with distal humeral fractures were treated with open reduction and internal fixation with DHP. There were 3 males and 15 females, ranging in age from 62 to 83 years (averaged, 71.4 years). According to AO classification, 1 patient was Type A2, 3 patients were Type A3; 2 patients were Type B1, 2 patients were Type B2; 3 patients were Type C1, 4 patients were Type C2, 3 patients were Type C3. The surgical approaches were either bilateral or via olecranon process of ulna. Early mobilization was initiated after surgery. Functional results were evaluated according to the Mayo elbow performance score (MEPS).

RESULTSThe average duration of follow-up was 14.6 months (ranged from 11 to 24 months). All the patients had a complete healing of their fractures. There was no infection, hardware failure or loss of reduction after the operations. The average Mayo elbow performance score (MEPS) was (92.2 +/- 9.6) (65 to 100 points), among which the score of pain was (42.5 +/- 5.8) (30 to 45 points), the score of motion range was (19.2 +/- 1.9) (15 to 20 points), the score of stability was (9.2 +/- 1.9) (5 to 10 points), and the score of function was (21.4 +/- 2.9) (15 to 25 points). According to MEPS, 11 patients got excellent results, 5 good and 2 fair.

CONCLUSIONTreatment of distal humeral fractures in elderly osteoporotic patients with AO distal humeral plates can obtain immediate stabilization which facilitates early mobilization of the elbow.

Aged ; Aged, 80 and over ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; surgery

Animals ; Gene Expression ; Genetic Therapy ; Male ; Mice ; Spermatogenesis ; Stem Cell Transplantation ; Testis ; metabolism ; Transfection

OBJECTIVETo construct recombinant baculoviruses co-expressing three structural genes vp2, vp6 and vp7 of rotavirus, and assemble rotavirus-like particles (VLPs) in BmN cells.

METHODSHuman group A rotavirus was cultivated in MA104 cells, and the RNA was extracted and the three genes were obtained by RT-PCR. The PCR products were inserted into the transfer vectors pFBDM and pUCDM, respectively. A enhanced green fluorescent protein gene (egfp) driven by IE1 promoter was introduced into pFBDM to investigate the efficiency of infection. The expression baculoviruse was constructed by Tn7 and Cre-LoxP recombinant and transfected into BmN cells. The gene expression was determined by detecting 6-His tag fused into VP7 C-terminus, and the assembled VLPs were observed by transmission electron micrography.

RESULTSThree genes of rotavirus were cloned and BmMultiBac was constructed. The genes were expressed and the rotavirus-like particles assembled in BmN cells successfully as verified by ELISA and electron microscope.

CONCLUSIONWe have successfully constructed the recombinant baculovirus co-expressing the 3 structural genes of rotavirus, which provide the basis for producing protein complex containing multiple subunits and investigation of the structure of the macromolecules.

Animals ; Baculoviridae ; genetics ; metabolism ; Bombyx ; virology ; Capsid Proteins ; genetics ; Cell Line ; Gene Expression ; Genetic Vectors ; Rotavirus ; genetics ; metabolism

OBJECTIVETo evaluate the efficacy and safety of endovascular aneurysm repair (EVAR) for abdominal aortic aneurysm (AAA), and to compare the prognosis between patients of different ages.

METHODSThe hospitalization and follow-up data of 81 AAA patients treated by EVAR from May 2005 to May 2011 were retrospectively analyzed. All the patients were divided into advanced age group (age ≥ 75 years, 24 cases) and relatively young group (age < 75 years, 57 cases). General conditions, comorbidity, procedure, in-hospital complications, and follow-up were compared between these two groups.

RESULTSAll covered stents were successfully deployed, a technical success rate of 91.4% (74/81) was achieved. There was no intraoperative death. In-hospital mortality was 1.2% (1/81). The follow-up rate was 91.4% (74/81), with a mean follow-up of 47.5 months. Twelve deaths were recorded during follow-up, 1, 2, 3, 4, and 5-year survival rates were 98.6%, 92.2%, 80.8%, 58.7%, and 44.1%, respectively. When compared with relatively young group, the advanced age group had a lower rate of abdominal pain as the major symptom, but a higher rates of renal diseases and coronary artery diseases. Furthermore, the advanced age group had a longer stay in intensive care unit and higher morbidity of endoleaks, and also tended to have increased rates of pulmonary infection and access site hematoma, while the other parameters were similar between the two groups.

CONCLUSIONSEVAR of AAA is less invasive, safe, and effective during short to mid-tern follow-up. The patients of advanced age suffer from higher rates of some complications, thus careful perioperative preparation and intensive monitor are mandatory for preventing or treating potential complications and improving prognosis for these patients.

Aged ; Aged, 80 and over ; Aortic Aneurysm, Abdominal ; surgery ; Blood Vessel Prosthesis Implantation ; adverse effects ; methods ; Endoleak ; etiology ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Prognosis ; Retrospective Studies ; Survival Rate ; Treatment Outcome