Alfimeprase(ALF)is a recombinantly modified variant of non-hemorrhagic zinc metalloproteinase fibrolase.The target gene alf was obtained from the clone vector p43-alf and cloned into the Pichia pastoris expression vector pPICZ? A.Through high efficiency transformation and Zeocin selection,the recombinant strains of pPICZ?A-alf /GS115 were isolated.In order to achieve a high level expression of recombinant Alfimeprase(rALF),optimization of pH value,methanol daily addition concentration,cell density and methanol induction time points were carried out,and the production of rALF reached up to 425 mg/L.By His?Bind chromatography,the purity of secreted rALF was as high as 95 %.SDS-PAGE and Western blot analysis show that rALF has a molecular weight of about 24 kDa and is bound specifically to anti-His?tag monoclonal antibody.Activity identification results of the modified fibrin plate method demonstrate that the secreted rALF has high fibrinolytic activity.Thus sets up an optimized expression system for ALF,which will play an important role in its further studies and industrial production.

Objective To investigate the differences of apparent diffusion coefficient(ADC)value in normal breasts and to evaluate the correlation between ADC value and corresponding histology.Methods Sixty-two normal breasts including 42 normal breasts of 42 patients with unilateral lesions and 20 normal breasts of 10 volunteers were studied.The ADC value of all 62 normal breasts were calculated when b value was given from 1000 to 0 s/mm~2,1000 to 500 s/mm~2and 500 to 0 s/mm~2.The MRI features of 60 normal breasts were classified into 3 types(dense,lobular-speckled,degenerative types)according to Wolf's classification and histology.Results DWI and ADC images were different in 3 types of normal breasts because of different histologic structures.The mean ADC value of the dense type breasts was(1.70? 0.37)?10~(-3)mm~2/s,the lobular-speckled type was(1.93?0.46)?10~(-3)mm~2/s and the degenerative type was(1.18?0.65)?10~(-3)mm~2/s(F=12.998,P=0.000).There were no significant differences between the dense type and the lobular-speckled type(F=2.167,P=0.147),but significant differences between the dense type and the degenerative type,the lobular-speckled type and the degenerate type(F=5.593 and 19.128;P=0.029 and 0.000).When b value decreased,the ADC value of the dense type and the lobular- speckled type increased correspondingly,but the degenerative type didn't increase apparently.Conclusion ADC value was influenced by histologic structures in normal breasts and also was influenced by b value in the dense type and lobular-speckled type breasts.

OBJECTIVETo explore the effect of Yanghe Huayan Decoction (YHD, a representative recipe for warming yang mass dissipating) in inhibiting precancerosis of breast cancer (PBC) and on the protein and mRNA expression of ki67.

METHODSThe PBC rat model was established by dimethylbenzanthracene (DMBA), and 9 weeks later rats were randomly divided into the blank control group, the model group, the YHD group, the Sanjie Huatan Decoction group (SHD), the Pingxiao Tablet group (PT), and the tamoxifen group. Rats in the model group were administered with water by gastrogavage. Rats in the YHD group received YHD (deglued antler powder 12 g, prepared rhizome of rehmannia 9 g, cassia bark 6 g, white mustard seed 3 g, zedoary root 12 g, appendiculate cremastra pseudobulb 15 g, chekiang fritillary bulb 9 g, licorice root 6 g) at the daily dose of 7.2 g/kg by gastrogavage. Rats in the SHD group received SHD (oldenlandia diffusa 15 g, Scutellaria Barbata 15 g, Trichosanthes Kirilowii 15 g, pinellia 9 g) at the daily dose of 5.4 g/kg by gastrogavage. Rats in the PT group received PT at the daily dose of 144 mg/kg by gastrogavage. Those in the tamoxifen group received tamoxifen at the daily dose of 4 mg/kg by gastrogavage. Pathomorphological changes of the breast tissue were observed by HE staining. The positive rate and the gray value of ki67 expression were detected by immunohistochemical assay. And the expression of ki67 mRNA was detected by q-PCR.

RESULTSCompared with the model group, the general hyperplasia and the occurrence rate of precancerous lesion were higher and the occurrence rate of invasive carcinoma was lower in each treatment group (P < 0.05). Except the SHD group, the intensity of ki67 grey value increased in each treatment group (P < 0.05, P < 0.01). Except the PT group, the positive rate of ki67 and mRNA expression of ki67 increased in the rest treatment groups (P < 0.05, P < 0.01). Compared with the YHD group, there was no statistical difference in the occurrence rate of infiltration or the occurrence rate of precancerous lesion (P > 0.05). The positive rate of ki67 expression and mRNA expression of ki67 increased in the PT group, showing statistical difference (P < 0.05).

CONCLUSIONSYHD could partially inhibit and reverse canceration of breast cancer. It also could inhibit ki67 protein and mRNA expression. Its effect was similar to tamoxifen and superior to PT. So it was suitable for prevention and treatment of precancerous lesion of breast cancer.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ki-67 Antigen ; metabolism ; Mammary Neoplasms, Experimental ; chemically induced ; drug therapy ; metabolism ; Precancerous Conditions ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley

This paper introduces an 16-lead digital EEG signal acquisition system, which applies MCU MSP430 as central control unit with high performance analog devices and high speed multi-channel, multi-bit analog-to-digital converter as peripheral to retrench analog circuit. Data is transferred to PC by USART interface. Software on PC based on virtual instrument technology realizes real-time detection, display and storage. The system has many advantages such as high precision, stable performance, small volume and low power dissipation, thus provides a new means for digital EEG signal acquisition.
Analog-Digital Conversion ; Electroencephalography ; instrumentation ; Equipment Design ; Signal Processing, Computer-Assisted ; instrumentation ; Software

Adult ; Humans ; Male ; Middle Aged ; Neoplasms, Neuroepithelial ; radiotherapy ; surgery

Dental age plays an important role in age estimation. It has often been used together with skeletal age to improve the accuracy of age estimation abroad, but seldom performed in China. As a noninvasive technology, dental radiological imaging has been widely used in age estimation. By observing the age-related changes such as the pulp cavity and development of crown and root on radiographs. Gleiser and Hunt, as well as Demirjian have developed different methods to determine the age of human. Demirjian's method has been proved to be more accurate but with limitation when used in persons of eighteen and above. The accuracy and reliability of the measurements on pulp cavity could be improved as the development of computed tomography with its high resolution and intelligent software. As a convenient and accurate method, age estimation from dental computed tomographs would be more promising in the future for forensic scientists and anthropologists.
Age Determination by Teeth/methods* ; Dental Pulp/diagnostic imaging* ; Forensic Dentistry ; Humans ; Radiography, Panoramic/methods* ; Tomography, X-Ray Computed/methods*

BACKGROUNDIt has been confirmed that the concentration of cefepime in cerebrospinal fluid (CSF) could reach the 10% of its concentration in plasma, exceeding the inhibitory concentration to 90% of organisms (MIC(90)) for common bacteria. However, the blood-brain barrier (BBB) penetration ability of cefepime is still unclear. The aim of this study was to measure the CSF concentration of cefepime in patients after neurosurgical operations, and to determine the penetration of the drug through an incomplete BBB.

METHODSEight patients who received ventricular drainage (VD group) and 5 who underwent lumbar puncture drainage (LPD group) were enrolled into this study. Cefepime (2 g) was injected intravenously in 30 minutes after the neurosurgeries. The concentrations of cefepime in the CSF and plasma were measured by high-pressure liquid chromatography (HPLC) at different time points.

RESULTSThe CSF concentrations of cefepime at different time points in the VD group were significantly higher than those in the LPD group (P < 0.05). In the VD group, the concentration of cefepime in CSF reached the peak ((22.54 +/- 14.06) microg/ml) at 1 to 2 hours after the injection, while in the LPD group at 4 hours ((5.61 +/- 3.73) microg/ml). In both groups, the peak was higher than the MIC(90) of most common bacteria in intensive care unit. The ratio of CSF to plasma cefepime concentrations ranged from 0.30 to 2.14 in the VD group and 0.03 to 1.14 in the LPD group.

CONCLUSIONAfter neurosurgeries, CSF concentration of cefepime can reach a therapeutic level. Thus, the drug could be used to prevent and treat postoperative intracranial infection.

Anti-Bacterial Agents ; cerebrospinal fluid ; Blood-Brain Barrier ; Cephalosporins ; cerebrospinal fluid ; Humans ; Neurosurgical Procedures

Child, Preschool ; Female ; Histiocytosis, Non-Langerhans-Cell ; diagnosis ; etiology ; therapy ; Humans

OBJECTIVETo explore changes of Clara cell protein (CC16) and surfactant protein-D (SP-D) in the serum of patients with silicosis.

METHODThe concentrations of CC16 and SP-D were measured in the serum by sandwich enzyme-linked immunosorbent assays. The subjects consisted of 30 healthy volunteers and 90 silica-exposed workers including silica-exposed group, the silicosis of suspects group (0(+)) and the silicosis phase I group, 30 subjects each groups.

RESULTSThe concentrations of CC16 in the serum was significantly decreased in silica-exposed workers compared to controls (P < 0.01); The concentrations of CC16 in the serum were higher in lifelong nonsmokers than the current smokers in control subjects (P < 0.05), but they were no differences between lifelong nonsmokers and current smokers of 90 silica-exposed workers. Compared with control subjects, the levels of SP-D in the serum of silicosis suspects (0(+)) and silicosis phase I groups were significantly elevated (P < 0.01, respectively), which were also higher than silica-exposed group (P < 0.05 and P < 0.01, respectively), Discriminant equations set by CC16 and SP-D were used in diagnosis of silicosis, and the rate of accuracy in healthy volunteers, the silica-exposed group and the silicosis phase I group were 86.7%, 86.7% and 76.7%, respectively, The total rate of correct classification hit 84.2%.

CONCLUSIONThe serum CC16 of long-term silica-exposed workers is decreased, and SP-D is increased gradually.

Adult ; Case-Control Studies ; Epithelial Cells ; metabolism ; Humans ; Male ; Middle Aged ; Pulmonary Surfactant-Associated Protein D ; blood ; Silicosis ; blood ; Uteroglobin ; blood

OBJECTIVETo determine whether ischemia-induced bone marrow-derived EPCs mobilization is impaired in diabetic mice and the association with vascular endothelial growth factor (VEGF) release post ischemia.

METHODSC57Bl/6 mice were injected with 40 mg x kg(-1) x d(-1) streptozotocin for 5 days to induce diabetes and mice with fasting glucose > 10 mmol/L were included to DM group, control mice were injected with placebo. Two months later, hindlimb ischemia was induced by left femoral artery dissection and ligation. The ischemia was visualized by tetrazolium dye staining and pre-mortem angiography. The percentage of c-Kit(+)/Sca-1(+)/flk-1(+) early EPCs in peripheral blood mononuclear cells (PBMCs) was detected by flow cytometric analysis on days 0 (pre-ligation), 1, 3, 5, 7 and 14 days post-ligation. The plasma VEGF level was measured with a standardized ELISA-Kit.

RESULTSCirculating EPCs number was significantly lower in diabetic mice than that in control mice (0.60% +/- 0.03% vs. 0.95% +/- 0.09%, P < 0.001) and the plasma VEGF was undetectable in all animals before ligation Similar EPCs kinetics following induction of hindlimb ischemia were shown in both groups. However, EPCs mobilization was significantly impaired in diabetic mice compared with control mice within 3 days post ischemia (day 1: 1.16% +/- 0.20% vs. 1.80% +/- 0.32%, P < 0.05; day 3: 1.38% +/- 0.34% vs. 2.37% +/- 0.52%, P < 0.05). In parallel, plasma VEGF increase post ischemia was significantly less in diabetic mice than that in control mice (day 1: 73.1 pg/ml +/- 18.6 pg/ml vs. 128.5 pg/ml +/- 44.2 pg/ml, P < 0.05).

CONCLUSIONOur data suggest that ischemia-induced bone marrow-derived EPCs mobilization is impaired in diabetic mice, which may be related to the insufficient release of plasma VEGF post ischemia.

Animals ; Bone Marrow Cells ; cytology ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; physiopathology ; Hematopoietic Stem Cell Mobilization ; Ischemia ; metabolism ; physiopathology ; Mice ; Mice, Inbred C57BL ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism