OBJECTIVETo screen and characterize the short peptides which bind specifically to interleukin-2 (IL-2) receptor alpha chain (IL-2Ralpha) for acquisition of small antagonists for blocking the binding of IL-2 with IL-2Ralpha.

METHODS12-mer phage displayed peptide library was screened with the target cells of MT-2 cells which expressed IL-2Ralpha at high levels. The binding phage clones were eluted by anti-IL-2Ralpha monoclonal antibody. After 3 rounds of screening, the positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, and the amino acid sequences of the positive clones were deduced from the DNA sequences.

RESULTSSeven positive clones were screened out of the 17 phage clones bound to MT-2 cells. The positive clone M15 could bind specifically to MT-2 cell and PHA-activated peripheral blood monouclear cells. Amino acid sequence analysis identified 6 sequences, all of which contained hydrophilic residues, and 5 of these 6 sequences included Tyr, Phe and Leu conservative residues.

CONCLUSIONThe peptide sequences containing Tyr, Phe conservative residues identified in this study can bind to cell surface IL-2Ralpha.

Amino Acid Sequence ; Cell Line, Transformed ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Peptide Library ; Peptides ; genetics ; metabolism ; Protein Binding ; Receptors, Cell Surface ; metabolism ; T-Lymphocytes ; cytology ; metabolism

OBJECTIVETo study the clinical efficacy and adverse reaction of comprehensive therapy mainly with Chinese anti-cancer medicinal perfusion/embolization and assisted with Chinese drug-therapy based on TCM Syndrome Differentiation in treating primary liver cancer.

METHODSForty-one patients with liver cancer were divided into the treated group and the control group. To the treated group turmeric oil microballoon, cinobufotalin, Aidi injection and iodized oil were given via hepatic artery perfusion/embolization, and to the control group chemotherapeutic agents and iodized oil were given for instead. Besides, both groups were given Chinese herbs according to TCM Syndrome Differentiation additionally.

RESULTSThe tumor inhibitory rate in the treated group and the control group was 77.78% and 69.57% respectively, with insignificant difference between them. The improvement of fatigue and anorexia in the treated group was better than that in the control group (P < 0. 05). The 6-month, 12-month and 24-month survival rate in the treated group and control group was 61.11% vs 56.62%, 27.78% vs 30.43% and 22.22% vs 26.09%, respectively, the difference between the two groups was insignificant. The occurrence of adverse reactions such as decreasing of white blood cells, platelet and hemoglobin, nausea and vomiting were obviously lower in the treated group than those in the control group (P < 0.05).

CONCLUSIONChinese anti-cancer medicinal perfusion/embolization has affirmative short-term clinical effect in treating primary liver cancer with few adverse reactions, which was tolerable to patients, but its long-term clinical efficacy needs further observation.

Adult ; Aged ; Antineoplastic Agents, Phytogenic ; administration & dosage ; Carcinoma, Hepatocellular ; drug therapy ; therapy ; Chemoembolization, Therapeutic ; methods ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Liver Neoplasms ; therapy ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy

Multiple displacement amplification (MDA) is a new technology for whole genome amplification (WGA), which can generate large amount of high-quality DNA and features high amplification efficiency and fidelity. MDA combined with conventional PCR techniques has been successfully applied for preimplantation genetic diagnosis, which has broaden latter's clinical applications.
Genome, Human ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Polymerase Chain Reaction ; methods ; Preimplantation Diagnosis ; methods

To investigate the interaction between tumor necrosis factor alpha (TNF alpha) mimotopes and TNF alpha-binding peptides screened from random phage display peptide library with TNF alpha mimotopes displayed on phage clone as the target, the computational docking program AutoDock (with confirmation calculations using Discover) was used to predict and analyze the binding modes of LLT-18 (TNF alpha binding peptide, sequence EHMALTYPFRPP) with TNF alpha, after which LCS-7 (TNF alpha mimic phage clone, displayed positive sequence c-RRPAQSG-c) was docked to LLT-18 manually. The binding between LLT-18 and TNF alpha or LCS-7 was stabilized predominantly through electrostatic interaction and H-bond formation. The Arg residues in TNF alpha or LCS-7 were important for their interaction with LLT-18. For LLT-18, the key amino acid residues were Glu1, His2, Met3 and Tyr7. These results suggest the feasibility of screening ligand to single epitope with specific phage clone as the target, and of predicting the interaction between small peptides by computer-aided molecular modeling.
Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; immunology ; Biotinylation ; Computer Simulation ; Drug Evaluation, Preclinical ; methods ; Epitopes ; immunology ; Humans ; Ligands ; Models, Molecular ; Oligopeptides ; chemistry ; metabolism ; Peptide Library ; Protein Conformation ; Solubility ; Tumor Necrosis Factor-alpha ; chemistry ; immunology ; metabolism

BACKGROUND: Polymethylmethacrylate (PMMA), commonly known as bone cement, has been widely used in the orthopedic surgery. It ensures the immediate stability of prosthesis and the minimal micromotion at the cement-bone interface, allowing early weight-bearing after surgery. OBJECTIVE: To investigate the biomechanical performance of Sanders type III fracture of the calcaneus by using PMMA bone cement as a treatment. METHODS: Eight adult cadaveric ankle and calcaneus specimens were selected and served as normal controls after detection of biomechanical properties. Another eight specimens were collected and randomized into experimental group and control group to make a model of Sanders type III fracture in the calcaneus. In the experimental group, PMMA bone cement was injected into the defect area. In the control group, the artificial bone was implanted in the defect area and a steel plate was used to fix the lateral calcaneus. Biomechanical properties of the specimens in the experimental and control group were detected. RESULTS AND CONCLUSION: (1) Strain and stress of the calcaneus: The stress distribution of the calcaneus in the normal control group was consistent with that in the experimental group, and there was no significant difference between the two groups. The stress of the calcaneus in the experimental group was similar to that in the control group with no significant difference. (2) Displacement and axial stiffness of the calcaneus: Compared with the normal control group, the calcaneal displacement in the experimental group only decreased slightly, and there was no significant difference between the two groups, and likewise, the calcaneal displacement in the control group increased slightly. In the experimental group, the axial compression strength was (21.98±1.88) MPa and the axial compression stiffness was (1 633±150) N/mm. Therefore, there was no significant difference between the experimental group and the normal control group (P > 0.05). (3) Contact strength of the subtalar joint: Fractures basically recovered with good outcomes after PMMA bone cement injection. To conclude, by using PMMA bone cement in the treatment of calcaneus fractures, the scientific validity and clinical utility can be ensured.

OBJECTIVETo evaluate the mid-term clinical outcome of endoscopic greater saphenous vein harvesting (EVH) in coronary artery bypass grafting (CABG). Method A total of 205 patients receiving off-pump CABG between July, 2012 and April, 2013 at our department were enrolled in this study, including 66 patients (35 male and 31 female patients with a mean age of 60.3±7.92 years) undergoing EVH and 139 patients (109 male and 30 female patients with a mean age of 59.20±8.37 years) undergoing open greater saphenous vein harvesting (OVH).

RESULTSThe surgical procedures were completed smoothly in all the cases. The perioperative mortality rates was 3.03% (2/66) in EVH group, as compared with 3.60% (5/139) in OVH group (P=1.00). Acute myocardial infarction (AMI) occurred during the perioperative period in 3 (2.16%) patients in OVH group and in 1 (1.52%) patient in EVH group. Perioperative low cardiac output syndrome was diagnosed in 4 (2.88%) patients in OVH group and in 2 (3.03%) in EVH group (P>0.05). During the follow-up, 8 (8.80%) patients in OVH group and 5 (8.06%) in EVH group had recurrent angina (P=0.93). No patients experienced AMI during the follow-up. The 2-year patency rate of the venous grafts was 83.59% in OVH group and 82.22% in EVH group (P=0.73).

CONCLUSIONEVH has significant advantage in reducing the complications of the incision in the lower limbs. The mid-term patency rates of venous grafts are similar between OVH and EVH, but the long-term patency rate needs further evaluation.

Aged ; Coronary Artery Bypass ; Endoscopy ; Female ; Humans ; Lower Extremity ; Male ; Middle Aged ; Saphenous Vein ; transplantation ; Tissue and Organ Harvesting ; Vascular Surgical Procedures

OBJECTIVETo observe the impact of specific short hairpin RNA (shRNA) targeting survivin gene on tumorigenesis and angiogenesis of human brain glioblastoma U251 cells in vivo of nude mice.

METHODSU251 cells, U251-SR cells transfected stably with shRNA eukaryotic expression vector pWH1-SR targeting survivin gene, and U251-P cells transfected stably with blank pWH1 vector, were inoculated respectively into subcutaneous tissue in flank of 15 nude mice (each group 5 mice), and the tumor growth status was observed and measured. Protein expressions of survivin, proliferating cell nuclear antigen (PCNA) and factor VIII related antigen (F VIII RAg) were investigated by immunohistochemistry SABC method, apoptotic cells were screened by TUNEL method, furthermore proliferative index (PI), apoptotic index (AI) and microvessel density (MVD) were measured respectively in each group of tumor specimens.

RESULTSComparing with those in U251 and U251-P groups, in U251-SR group, the tumorigenesis time delayed, tumor grew slowly, both tumor volume and tumor weight decreased significantly (P < 0.01 for both); Survivin protein expression was down-regulated markedly; PI and MVD decreased significantly, whereas AI increased remarkably (P < 0.01 for all).

CONCLUSIONSThe specific shRNA targeting survivin gene can inhibit significantly tumorigenesis and angiogenesis of U251 cells in vivo.

Animals ; Apoptosis ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Glioblastoma ; metabolism ; pathology ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Mice ; Mice, Nude ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Transplantation ; Neovascularization, Pathologic ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Repressor Proteins ; Transfection

OBJECTIVETo investigate the prevalence of psychiatric disorders in a representative sample of primary and middle school students in Hunan Province.

METHODSA total of 9 495 children aged 5-17 years from Hunan urban and rural schools were enrolled by a cluster sampling and a two-phase design. The students' psychiatric status was assessed using the Investigation Screening Inventory for Child Mental Disorder and a semi-structured interview designed based on the DSM-IV criteria.

RESULTSThe overall prevalence of psychiatric disorders was 16.22%. Attention-deficit and disruptive behavior disorders were the commonest in the diagnostic categories of psychiatric disorders (10.69%). Regarding specific disorders, the most prevalent was attention-deficit/hyperactivity disorder (5.95%). Psychiatric disorders were more prevalent in boys than in girls (20.49% vs 11.16%; p<0.01). The prevalence of attention-deficit and disruptive behavior disorders in boys was higher than in girls (14.76% vs 5.87%; p<0.01). The prevalence of psychiatric disorders in middle school students (12-17 years) was significantly higher than in primary students (5-11 years) (18.38% vs 14.64%; p<0.01). There were no significant differences in the prevalence of psychiatric disorders between urban and rural students.

CONCLUSIONSPsychiatric disorders are common among primary and middle school students in Hunan Province. The prevalence of this disorder in boys is higher than in girls. The middle school students have higher prevalence than primary students.

Adolescent ; Age Factors ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Mental Disorders ; epidemiology ; Prevalence ; Sex Factors

Hepatitis E is an acute hepatitis casused by hepatitis E virus (HEV) in developing countries, where it occurs as cases sporadic and in epidemics form. The causative agent, hepatitis E virus, is transmitted primarily by the fecal-oral route. HEV is icosahedron non-enveloped virus, and its genome is a single-stranded, positive-sense, 3'-polyadenylated RNA about 7.5 kb in length. It contains three open reading frames (ORFs). Of which ORF1 codes for a polyprotein of 1693 amino acids and contain domains homologous to a viral methyltransferase, a papainlike cysteine protease, an RNA helicasre, and an RNA-dependent RNA polymerase, besides the most hypervariable region of the HEV genome. And ORF3 codes for a 123-amino-acide-long polypeptide with unknown function. While the major viral capsid protein (pORF2, ORF2 codes) of 660 amino acid was showed to contain the protective epitope. The bacterially expressed polypeptide disignated as NE2 has been proved to be a protective antige. And the anti-NE2 monoclonal antibodies (mAb) was screend, two of these mAbs 8C11 and 8H3 were showed to be against separate conformational neutralization epitope of hepatitis E virus (HEV). And these two mAb were used to screen for binding peptides from a 7-peptides phage display library. After four rounds of panning, tweenty-one positive monoclonal phages (11 for 8C11, and 10 for 8H3) were selected and the inserted fragments were sequenced. The DNA sequence coding for the obtained dominant peptide 8C11 (N'-His-Pro-Thr-Leu-Leu-Arg-Ile-C', named 8C11A) and 8H3 (N'-Ser-Ile-Leu-Pro-Tyr-Pro-Tyr-C', named 8H3A) were then synthesized and cloned to insert between amino acid 78 to 83 of hepatitis B core antigen (HBcAg), then expressed in E. coli. The recombinant proteins aggregate into homodimer or polymer on SDS-PAGE, and could bind with mAb 8C11 and 8H3 in Western blotting. Respectively, the recombinant protein C8C11A showed to be dimer mainly, which can bind with mAb 8C11. The monomer and dimer of C8H3A are in the same amount on SDS-PAGE, but only the dimer could bind with mAb 8H3 on Western blotting. The renatured recombinant proteins were all showed to aggregate into virus like particles which were similar as HBcAg on transmission electron micrograph. The dominant peptide 8H3A (N'-Ser-Ile-Leu-Pro-Tyr-ProTyr-C') that selected out by mAb 8H3 was further chemo-synthesized, and its binding activity was confirmed by BIAcore biosensor. The result showed that this 7-peptide can bind with mAb 8H3 in a big Ka and Kd form, which means the binding is not stable. These results implicated that conformational dependent neutralization epitope could be partially modeled by short peptide, which provided a feasible route for subunit vaccine development.
Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Epitopes ; chemistry ; genetics ; immunology ; metabolism ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis E virus ; genetics ; immunology ; metabolism ; Mice ; Microscopy, Electron ; Molecular Sequence Data ; Peptide Library ; Peptides ; chemistry ; genetics ; immunology ; metabolism ; Recombinant Proteins ; genetics ; immunology ; metabolism ; ultrastructure ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics ; immunology ; metabolism ; ultrastructure

OBJECTIVETo establish a bone metastasis model of prostate cancer by intratibia injection of Du145 in nude mice, observe the local growth of tumor in tibia and then assess application value of this model.

METHODSFor 9 male nude mice, Du145 (5 x 10(6)) was injected in tibia by a TB syringe with a 29-gauge needle at a dose of 30 microl per mouse. Then the vital signs of the nude mice were observed. When the mice were dying, they were sacrificed, and the tissues of right hindlimbs, lymphatic nodes, lungs and livers were taken out, fixed in 10% formalin, embedded in paraffin, stained by HE and then observed microscopically.

RESULTSIncidence of bone tumor after intratibia injection was 67% (6 out of 9). About 48 days later, there were some small palpable nodes in right hind-limbs of the 6 mice and they couldn't walk normally. About 55 days later, cachexia occurred in them. After dissection, some carrion-like tissue grew from marrow cavity to muscular spatium, which was identified as tumor tissue by HE. The envelop of livers became crampy, and acute hepatitis could be diagnosed through microscopy, which represented a large scale of hepatocytic death, liver sinus dilatation and hyperemia, hepatic lobule infiltrated by lymphocyte, macrophage and inconspicuous hyperplasia. Since hypohepatia occurred too early, we couldn't detected distant metastases.

CONCLUSIONThe intratibia injection model is an optimal animal model to study metastasis of prostate cancer. It mimics the natural situation of human prostate cancer and will help to understand the mechanisms of androgen-independence and osseous metastasis, and tumor-host determinants of PSA expression.

Animals ; Bone Neoplasms ; secondary ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Prostatic Neoplasms ; pathology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays ; methods