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 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 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

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

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

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.

Objective To explore the brain natriuretic peptide (BNP) on prognostic value in patients with viral myocarditis.Methods A 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.Results The 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.Conclusion Levels 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.

Objective To evaluate the value of heart rate variability (HRV ) and sympathetic skin response (SSR) in differential diagnosis of Parkinson's disease (PD) .Methods We selected 36 PD ,17 essential tremor (ET) and 59 Parkinsonism-Plus (PD-plus) patients diagnosed in our department .We recorded HRV during the period of quiet breath ,deep breath and Vasaval as well as SSR incubation period of the upper limbs and lower limbs . Results ① During the period of quiet breath ,HRV decreased significantly in PD group compared with that in ET group (P< 0 .05) ,and abnormality rate of HRV in PD group was higher than that in ET group .HRV did not significantly differ between PD and PD-plus .HRV did not significantly differ among the three groups during the period of deep breath and Vasaval .② SSR incubation period showed no significant difference among PD ,PD-plus and ET groups .③ The specificity and sensibility of HRV in PD and PD-plus differential diagnosis were 22 .8% and 82 .6% , while for PD and ET differential diagnosis the values were 66 .9% and 82 .6% . The specificity and sensibility of SSR in PD and PD-plus differential diagnosis were 39 .0% and 52 .6% , 29 .4% and 52 .6% , respectively ,for PD and ET differential diagnosis .Conclusion Compared with those in ET patients ,HRV in PD patients decreases and HRV abnormality rate increases .HRV is of great value in differential diagnosis of PD and ET but not in differential diagnosis of PD and PD-plus .Determining SSR incubation period does not have much value in differential diagnosis of PD ,PD-plus and ET .

Objective To explore the feasibility of fabricating a self-powered and implantable stimulator . Methods Based on pyroelectric effect and high penetrability of infrared ray , an implantable stimulator was designed and fabricated .The electrical performance of stimulator was measured under infrared ray illumination .We conducted the animal experiments in vitro and in vivo to observe the response of gastrocnemius contraction under stimulation .Results The stimulator could output the electrical signal under the periodical infrared ray illumination .The output voltage and current were proportional to the intensity of infrared ray ,which could reach up to 1 .2 V . A real-time electrical stimulation of frog gastrocnemius was conducted and obvious contraction was observed .The tension values increased with the increase of infrared intensity .We further carried out the in vivo experiment with a frog in order to evaluate the performance of the stimulator after being implanted into the body . The gastrocnemius would also be made to contract even though the infrared intensity decayed when through the skin .Conclusion The proposed pyroelectric stimulator can be self-powered and controlled through near-infrared illumination .This study can provide some guidance for solving the problems of implantable power .