OBJECTIVETo prepare the modified ZHER2V2 affibody with amino-terminal HEHEHE sequence and carboxyl-terminal GGGC sequence by gene recombinant expression,which is the basis for invasive HER2 imaging with affibody.

METHODSThe encoded affibody gene was optimized by codon preference of E. coli with gene designer software. The N-terminal of affibody was fused with HEHEHE sequence,while the C-terminal was fused with GGGC sequence. The synthetic gene was confirmed by Hind 3 endonuclease restriction and gene sequencing. The human epidermal growth factor receptor-2(HER2)affibody gene was sub-cloned into pET22b(+)plasmid and transformed into competent BL21(DE3)bacteria. The expression of modified affibody was induced with isopropyl Β-D-1-thiogalactopyranoside(IPTG)and identified by SDS-PAGE. The affibody was purified by nickel affinity binding and imidazole elution. The purified affibody was labeled with (68)Ga and its affinity was determined by saturation analysis with HER2-positive cells MDA-MB-361.

RESULTSThe affibody gene containing N-terminal HEHEHE and C-terminal GGGC sequences were confirmed by Hind 3 endonuclease restriction and gene sequencing. A newly expressed 8×10(3) protein was expressed from the induced recombinant bacteria identified by SDS-PAGE after sub-cloning HER2 affibody gene into pET22b(+)plasmid,transforming recombinant plasmid into competent BL21(DE3)bacteria and inducing the recombinant bacteria with IPTG. The expressed protein was purified from nickel agarose by 60 mmol/L imidazole eluting. The affinity Kd value of (68)Ga labeled affibody to HER2 positive MDA-MB-361 cells was 1.5 nmol/L.

CONCLUSIONThe affiibody ZHER2V2 containing N-terminal HEHEHE and C-terminal GGGC was successfully prepared by gene optimization,recombinant expression and affinity purification.

Affinity Labels ; isolation & purification ; Escherichia coli ; metabolism ; Gene Expression ; Humans ; Receptor, ErbB-2 ; genetics ; Recombinant Fusion Proteins ; biosynthesis

OBJECTIVETo observe the characteristics of the physiological uptake of uterus and ovaries on 18F-fluorodeoxyglucose positron emission tomography (FDG PET).

METHODSA total of 288 PET examinations performed in 247 women (164 with malignancies, 44 with benign diseases, and 39 without remarkable abnormality) were included for analysis, and clinical follow-ups were applied for at least 10 months to exclude pelvic diseases. The menstrual statuses, menstrual cycles, and related pelvic examinations with other modalities were inquired before each PET examination. PET scanning was performed from pelvis to neck with a Siemens ECAT EXACT HR + system. The uptake levels of uterus and ovaries were set as intense, moderate, and mild by comparing to liver uptake.

RESULTSIn 116 patients (131 examinations ) with regular menstruation, the endometrial uptake, usually in inverted cone shape surrounded by relatively low-uptake uterine wall, was observed with two peaks in the early menstrual flow phase and in the mid-cycle respectively; the ovarian uptake was more prominent in the mid-cycle, with the foci of uptake in ovoidal shape and located at the left and/or right side superior-posterior to the bladder. From the early menstrual flow phase to the late secretory phase of the menstrual cycles, the probabilities of mild uptake in both endometrium and ovaries were 7%, 86%, 80%, 58%, 20%, 40%, 64%, and 59%, respectively, indicating that the late menstrual flow phase and the early proliferative phase had the least probability of intense or moderate uptake. No intense uptake was observed in the 17 patients (19 examinations) presenting remarkably irregular menstrual cycle, 112 patients (136 studies) in menopause for 3 months to 39 years, and 2 patients without menstruation yet. Only one patient within 1 year of menopause and a 14-year-old girl expected to start menstruation showed mild to moderate uptake in the endometrium.

CONCLUSIONThe physiological endometrial and ovarian uptakes have specific shapes and positions on 18F-FDG PET images, which correlates well with the menstrual phases.

Adolescent ; Adult ; Female ; Fluorodeoxyglucose F18 ; pharmacokinetics ; Humans ; Menopause ; Menstrual Cycle ; Middle Aged ; Ovary ; diagnostic imaging ; metabolism ; Positron-Emission Tomography ; methods ; Uterus ; diagnostic imaging ; metabolism ; Young Adult

OBJECTIVETo develop a 18F-labeled amino acid, O-(2-[18F]fluoroethyl) - L-tyrosine(18F-FET), as a positron emission tomography (PET) tracer for imaging cerebral tumors.

METHODS18F-FET was synthesized. Preclinical studies including sterility, endotoxin, and toxicity tests were performed. Two brain tumor cases were studied using 18F-FET and compared with 18F-FDG.

RESULTSRadiochemical purity of 18F-FET was over 95% which remained stable for 6 hours. The 18F-FET injection was sterile and its endotoxin content accorded with the standards of Chinese Pharmacopoeia. The uptake of 18F-FET in the normal brain tissues was significantly lower than that of the tumor, and the images of the brain tumor were clearer than those of 18F-FDG.

CONCLUSION18F-FET can accumulate in the tumor tissues to give high quality images. It suggests that 18F-FET may be a safe and effective tracer for brain tumor imaging.

Adult ; Animals ; Brain Neoplasms ; diagnostic imaging ; Female ; Fluorine Radioisotopes ; Fluorodeoxyglucose F18 ; Glioblastoma ; diagnostic imaging ; Humans ; Male ; Mice ; Middle Aged ; Sarcoma 180 ; diagnostic imaging ; Tomography, Emission-Computed ; Tyrosine ; analogs & derivatives ; chemical synthesis

OBJECTIVETo find sensitive and specific micro-metastic markers for prostate cancer.

METHODSUsing nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people.

RESULTSThe expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage.

CONCLUSIONPSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.

Aged ; Antigens, Surface ; blood ; Biomarkers, Tumor ; blood ; Glutamate Carboxypeptidase II ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; pathology ; Prostatic Neoplasms ; blood ; pathology ; Tissue Kallikreins ; blood

OBJECTIVETo assess the correlation of O6-methylguanine-DNA methyltransferase (MGMT) to radiation sensitivity of nasopharyngeal carcinoma (NPC).

METHODSEighty randomly selected NPC patients were divided into high (+/++, n=62) and low (-/+/+, n=18) MGMT groups according to the results of MGMT detection using immunohistochemistry. All the patients received irradiation with external beam radiotherapy, and the radiation sensitivity of NPC was analyzed after the irradiation.

RESULTSThe rates of high and low radiation sensitivity were 83.3% and 16.7% in low MGMT group, respectively, showing significant differences from those of the high MGMT group (45.2% and 54.8%, respectively, chi(2)=4.393, P=0.036).

CONCLUSIONThe content of MGMT correlates to the radiation sensitivity of NPC and may serve as valuable indicators for predicting the radiation sensitivity of NPC.

Adolescent ; Adult ; Aged ; Carcinoma, Squamous Cell ; enzymology ; radiotherapy ; DNA Modification Methylases ; blood ; DNA Repair Enzymes ; blood ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; enzymology ; radiotherapy ; Radiation Tolerance ; physiology ; Tumor Suppressor Proteins ; blood ; Young Adult

OBJECTIVE: To determine the cellular compatibility of combined deproteinized bone(DPB) coated with hepatocyte growth factor (HGF), and to observe the adherent effect of osteoblasts in response to HGF. METHODS: Osteoblasts were isolated from fetal rabbits. Osteoblasts were cultured with DPB coated with HGF and deproteinized bone as experimental group and contral group, respectively. The proliferation and alkalinephosphatase activity were tested. Their growth was examined by inverted phase contrast microscope and scanning electronmicroscope. RESULTS: The osteoblasts were attached to the outside and inside surfaces and grew well. HGF/DPB could stimulate the alkalinephosphatase activity of the osteoblasts and improve the proliferation of the osteoblasts. CONCLUSION HGF/DPB has good biocompatibility and bone induction. HGF could improve the adherent effect of DPB on osteoblasts, and it could be used as scaffold material for the bone tissue engineering.
Animals ; Biocompatible Materials ; pharmacology ; Bone Substitutes ; metabolism ; Bone and Bones ; cytology ; Cell Proliferation ; Cells, Cultured ; Female ; Fetus ; Hepatocyte Growth Factor ; pharmacology ; Osteoblasts ; cytology ; Osteogenesis ; Pregnancy ; Rabbits ; Tissue Engineering ; methods ; Tissue Scaffolds

We constructed a recombinant vaccine of Mycobacterium tuberculosis rBCG-Rv2029c,and then identified it.Rv2029c antigen encoding gene was amplified by PCR.The enzyme digestion products were ligated into rpMV261-2029c recombinant plasmid,after double digestion of Rv2029c and pmv261 vector,and then we introduced the plasmid into BCG to construct rBCG-Rv2029c recombinant vaccine by electroporation method.Finally,we analyzed the expression of the recombinant protein by SDS-PAGE and Western blotting.A total of 1 020 bp Rv2029c gene successfully amplified by PCR was inserted into the plasmid pmv261,then the fusion gene was successfully transduced into BCG.After identified by double enzyme digestion,confirmed by gene alignment and by thermally induced with Western blotting,the recombinant protein had a free primary.The recombinant live vaccine of M.tuberculosis rBCG-Rv2029c is successfully constructed,which lay a foundation for the study of the immune mechanism of recombinant vaccine.

OBJECTIVETo explore the brain natriuretic peptide (BNP) on prognostic value in patients with viral myocarditis.

METHODSA total of 48 patients with viral myocarditis and 42 healthy people were enrolled and followed up for two years. The NYHA class and LVEF were recorded and the concentration of BNP were measured.

RESULTSThe concentration of BNP were higher and EF were lower in patients with viral myocarditis (P < 0.01) than contrast people. Higher levels of plasma BNP were related to higher mortality.

CONCLUSIONLevels of brain natriuretic peptide measured in the plasma could be a useful biochemical marker for the myocarditis, and high concentration of BNP may correlate with poor prognosis in patients with myocarditis.

Adolescent ; Adult ; Biomarkers ; blood ; Child ; Female ; Humans ; Male ; Middle Aged ; Myocarditis ; blood ; mortality ; physiopathology ; Natriuretic Peptide, Brain ; blood ; Prognosis ; Ventricular Function, Left ; Virus Diseases ; blood ; mortality ; physiopathology

OBJECTIVETo observe the effects of interleukin-8 monoclonal antibody on smooth muscle cell proliferation and balloon inflation-induced abdominal aorta stenosis in rabbits.

METHODSThirty-six New Zealand white rabbits were randomly assigned to balloon inflation group (group A, n = 12), interleukin-8 monoclonal antibody pre-treated rabbits (2 mg/kg for 3 days before balloon inflation, group B, n = 12) and sham-operated control group (group C, n = 12). Peripheral blood was collected before experiment and at 4 h, 1, 3, 7, 14, and 28 days post balloon inflation or sham operation and the levels of IL-8 were measured by enzyme linked immunosorbent assay (ELISA). The ratio of positive and negative masculine cells in the high power microscopic field was determined in proliferating cell nuclear antigen (PCNA) stained slide. Histopathologic examination was performed in abdominal aorta and luminal area, intima and tunica media area were measured.

RESULTSPlasma interleukin-8 began to rise at 4 h and peaked at 1 day and remained increased up to 28 days after balloon inflation in rabbits of group A, plasma interleukin-8 level in group A was significantly higher than in group B and C at 4 h and thereafter post operation. The ratio of positive and negative masculine cells was significantly increased in group A compared to group C and was significantly lower in group B than in group A. Abdominal aorta stenosis, luminal area, intima and tunica media area were significantly reduced in group B than in group A. Correlation analysis indicated that there were positive relations between plasma IL-8 level and intima thickness, area of intima and tunica media, respectively (r = 0.894, 0.783, 0.801, 0.912, all P < 0.01).

CONCLUSIONSPlasma IL-8 level is increased in this abdominal aorta stenosis model and is positively correlated to the severity of abdominal aorta stenosis. IL-8 monoclonal antibody could significantly reduce abdominal aorta stenosis in this abdominal aorta stenosis model.

Animals ; Antibodies, Monoclonal ; pharmacology ; therapeutic use ; Aorta, Abdominal ; pathology ; Aortic Coarctation ; drug therapy ; pathology ; Cell Proliferation ; drug effects ; Interleukin-8 ; immunology ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; Rabbits

AIMTo study the incidence of single nucleotide polymorphisms in ubiquitin-specific protease 26 (USP26) gene and its involvement in idiopathic male infertility in China.

METHODSRoutine semen analysis was performed. Infertility factors such as immunological, infectious and biochemical disorders were examined to select patients with idiopathic infertility. DNA was isolated from peripheral blood of the selected patients and control population, which were examined for mutations using polymerase chain reaction-single strand conformation polymorphism analysis. Furthermore, nucleotide sequences were sequenced in some patients and controls.

RESULTSOf 41 infertile men, 9 (22.0%, P = 0.01) had changes in USP26 gene on the X chromosome. A compound mutation (364insACA; 460G right triple arrow A) was detected in 8 patients (19.5%, P = 0.01) and a 1044T right triple arrow A substitution was found in 1 patient (2.4%, P > 0.05). All three variations led to changes in the coding amino acids. Two substitutions predict some changes: 460G right triple arrow A changes a valine into an isoleucine, and 1044T right triple arrow A substitutes a leucine for a phenylalanine. Another insertion of three nucleotides ACA causes an insertion of threonine. No other changes were found in the remaining patients and fertile controls.

CONCLUSIONThe USP26 gene might be of importance in male reproduction. Mutations in this gene might be associated with male infertility, and might negatively affect testicular function. Further research on this issue is in progress.

Asian Continental Ancestry Group ; ethnology ; genetics ; Case-Control Studies ; China ; Cysteine Endopeptidases ; genetics ; metabolism ; Endopeptidases ; genetics ; metabolism ; Humans ; Incidence ; Infertility, Male ; ethnology ; genetics ; Leydig Cells ; metabolism ; Male ; Polymorphism, Single Nucleotide ; genetics ; RNA, Messenger ; genetics ; metabolism ; Sertoli Cells ; metabolism ; Spermatogenesis ; genetics ; Testis ; metabolism ; Ubiquitin-Specific Proteases