Objective To investigate the effects of inositol hexaphosphate (IP6) on proliferation of human osteosarcoma cell line MG-63 and primary cultured osteoblasts so as to explore the optimal concentration for achieving anti-cancer effects.Methods We primary cultured and identified human osteoblasts.Then we made recovery and normal culture of human osteosarcoma cell line MG-63.We tested the proliferation of two kinds of cell lines under different concentrations of IP6 by MTT to determine the optimal concentration and then detected MG-63 cell cycle and apoptosis by flow cytometry.Results When IP6 concentration was more than 1 mmol/L,IP6 began to inhibit the proliferation of MG-63 cell line in the time-dose dependent manner.When the concentration reached 4 mmol/L,this inhibitory effect was the maximum.When IP6 concentration was 0.5 mmol/L or 1 mmol/L,the proliferation of osteoblasts was not obviously inhibited.When it was 2 mmol/L,the proliferation was slightly inhibited.A concentration of 4 mmol/L caused the apoptosis of osteoblasts.Conclusion IP6 can inhibit the proliferation of osteosarcoma cell line MG-63 and lead to its apoptosis.The optimal concentration is 2 mmol/L for achieving anti-cancer effects.

Angioplasty, Balloon, Coronary ; Coronary Artery Disease ; therapy ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; therapy ; Stents ; Vascular Malformations ; therapy

group Nor(P0.05).Conclusions The changes of ambulatory pulse pressure can reflect the degree of EH.The obvious increase of PP and decrease of DBP maybe are the exclusive features of ABPM in EH patients with target organs damage.

OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

Objective To measure the learning passion of medical students and evaluating its in-fluencing factors. Methods Taking 879 medical students as research subjects to conduct a questionnaire survey according to specialty and grade stratified sampling. The questionnaire contained two parts, includ-ing learning passion scale and general situation questionnaire. The effective recovery rate was 69.28%, 609 valid questionnaires were recovered. SPSS 22.0 and AMOS 21.0 software were used for statistical analysis of data, analyzing the reliability and validity of the questionnaire with internal consistency reliability coeffi-cient and confirmatory factor analysis. The factors of learning passion of medical students were analyzed by logistic regression analysis. Results The questionnaire of learning passion of medical students contained 12 measurement items, including 2 dimensions: harmonious passion and obsessive passion. The questionnaire was of fine reliability (Cronbach's Alpha=0.916) and validity (χ2/df=3.401,RMSEA=0.073,GFI=0.958). The
learning passion of medical students was at upper middle level (4.390±0.934). The influence of achievement level (OR=1.691, 95%CI=1.415 to 2.021), school satisfaction (OR=0.586, 95%CI=0.402 to 0.854) and profession plan (short-term plan OR=2.121, 95%CI=1.310 to 3.434;long-term plan OR=3.822,95%CI=1.972 to 7.405) on learning passion were statistically significant. Conclusion The questionnaire of learning pas-sion of medical students has fine reliability and validity. Achievement level, school satisfaction and profes-sion plan are factors affecting the learning passion of medical students.

Aged ; Aged, 80 and over ; Basilar Artery ; abnormalities ; Female ; Humans ; Male ; Middle Aged ; Trigeminal Neuralgia ; etiology

To express recombinant carboxypeptidase from Thermus aquaticus (Cpase Taq) in Pichia pastosis, the open reading frame coding thermostable Cpase Taq was optimized based on the preference of P. pastoris codon usage and synthesized in vitro. The novel gene was cloned into P. pastoris expression vector pHBM905A and the sequence coding 6xHis tag was fused with the ORF of Cpase Taq gene. The recombinant plasmid was named pHBM905A-Cpase Taq and transformed into P. pastoris GS 115. Transformants were induced with 1% methanol for 72 h until the enzyme yield reached 0.1 mg/ml. The enzyme was purified and its enzymatic properties were analyzed. The results showed that the specific enzyme activity reached maximum at 75 °C and pH 7.5, which was about 80 U/mg. It was the first report about the secretory expression of Cpase Taq in P. pastoris GS115. Because of its large-scale preparation, this enzyme may be applied in industrial hydrolysis of peptides into amino acids in the future.
Bacterial Proteins ; biosynthesis ; genetics ; Carboxypeptidases ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; Hydrolysis ; Open Reading Frames ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Thermus ; enzymology

BACKGROUND: Previous studies have demonstrated that neural stem cells play potential therapeutic effects on the repair of spinal cord injury. However, the time for acquiring the best allotransplantation effects remains unclear.OBJECTIVE: This study was designed to observe the repairing effects of allotransplantation of embryonic neural stem cells on the motor function of rat two posterior limbs after spinal cord injury and investigate the time effectiveness of the allotransplantation.DESIGN: A controlled observational experiment.SETTING: Laboratory of Molecular Biology, Dalian Medical University; Laboratory of Biomedicine, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, Liaoning Province, China.METHODS: This study was performed at the Laboratory of Molecular Biology, Dalian Medical University & Laboratory of Biomedicine, School of Environmental and Biological Science and Technology, Dalian University of Technology between July and August 2003. One albino rat of gestational 14-16 days was sacrificed for harvesting embryonic rat brain cells. Embryonic rat cerebral cortex and subcortical periventricular brain tissue were taken for in vitro culture of rat embryonic neural stem cells. An additional 30 adult Sprague Dawley rats were randomly divided into 3 groups with 10 rats in each group: control, early allotransplantation and delayed allotransplantation groups. All 30 rats were subjected to spinal cord transection injury, leading to rat paralysis of both lower extremities. Embryonic rat neural stem cells were transplanted into the rats in the early and delayed transplantation groups at 3 days and 3 weeks after injury, respectively. Following allotransplantation, motor function of rat two lower extremities was followed. At 4 weeks after allotransplantation of neural stem cells, rat spinal cord was harvested from transplanted region for immunohistochemistry in order to observe and compare the morphological change of rat spinal cord tissue among the 3 groups. The following protocol was performed in accordance with ethical guidelines stated in Guide for the use and care of laboratory animals, approved by the Committee on the Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources Commission on Life Scineces, National Research Council, China (1985).MAIN OUTCOME MEASURES: Motor functional recovery of rat two lower extremities after neural stem cell transplantation. Histomorphological change of rat spinal cord at 4 weeks after neural stem cell transplantation.RESULTS: Thirty rats were included in the final analysis. In the early and delayed transplantation groups, the motor function of rat two lower extremities was noticeably improved, in particular in the early transplantation group. In the two experimental groups, muscular strength of paralyzed rat two lower extremities began to recover 5 or 6 days after transplantation of neural stem cells. Two or three weeks later, all rats in the two experimental groups could crawl and four weeks later, two extremities could move actively (approximately approaching to score 3 prescribed as follows). In the control group, no recovery of paralyzed extremities was found. At 4 weeks after transplantation, in the early transplantation group, proliferative tissue could be visible in the spinal cord transplantation region. Through the use of microscope, a considerable number of new cells were found that presented with neuronal and glial cell-positive staining. In the control group, a cavity between two broken ends could be visible. Meanwhile, necrosis and vacuolar degeneration, and other symptoms in the stump of spinal cord were observed with a microscope. In the delayed transplantation group, the histomorphological change of spinal cord region was between the other two groups. No typical histomorphological change was found. A number of new cells were apparent with a microscope, but the number was less compared with the early transplantation group.CONCLUSION: Allotransplantation of embryonic neural stem cells promotes the recovery of rat motor function after spinal cord transection. Early transplantation acquires better therapeutic effects.

Objective To compare the position,volume and matching index (MI) of esophagus between quiet end-inspiration and end-expiration in three dimensional CT (3D-CT) assisted with active breathing control (ABC) and the corresponding phases in four dimensional CT (4D-CT).Methods Eleven patients with peripheral lung cancer underwent 4D-CT simulation scan and 3D-CT simulation scans in end-inspiratory hold (CTEIH) and end-expiratory hold (CTEEH) in succession.The 0％ phase was defined as end-inspiratory phase (CT0),while the 50％ phase was defined as end-expiratory phase (CT50).The proximal,mid-,and distal thoracic esophagus were delineated separately on CT0,CT50,CTEIH and CTEEH images.The position,volume and MI of each segment esophagus between CT0 and CTEIH,CT50 and CTEEH were compared.Results In the left-right (x) direction,the position differences in the proximal,mid-,and distal thoracic esophagus between CT0and CTEIH were (-0.02 ±0.16)cm,(0.06 ± 0.26)cm and (0.10 ± 0.33) cm respectively,and in the anterior-posterior (y) direction,the position differences were (0.04 ±0.24)cm,(0.04 ±0.12) cm and (0.08 ±0.15) cm respectively,and the position differences in the same direction were not statistically significant.In the x direction,the position differences of the proximal,mid-,or distal thoracic esophagus between CT50 and CTEEH were (-0.02 ±0.24) cm,(0.12 ± 0.37) cm and (0.26 ± 0.33) cm respectively,and in the y direction,the position differences were (0.03 ±0.21)cm,(0.04 ±0.17)cm and (0.14 ±0.18)cm respectively,and the position differences in x and y directions of proximal and mid-thoracic esophagus between CT50 and CTEEH were not statistically significant,while the position differences in x and y directions of distal thoracic esophagus between CT50and CTEEH were both statistically significant (t =0.025,0.024,P ＜ 0.05).The volumes of the proximal,mid-and distal thoracic esophagus were all larger in CT0and CT50 than those in CTEIHand CTEEH,but without statistical differences.The MIs of the volumes of the proximal,mid-and distal thoracic esophagus between CT0 and CTEIH were (0.50 ± 0.17),(0.50 ± 0.19) and (0.56 ± 0.08),respectively,and those between CT50and CTEEH were (0.50 ±0.16),(0.47 ±0.14) and (0.51 ±0.15),respectively.The MI of each segment esophagus between CT0and CTEIHwas larger than that between CT50 and CTEEH,but without statistical differences.Conclusions The influence of breathing modes on the centroid positions of the proximal,mid-thoracic normal esophagus were not significant and there were spatial mismatches for any segment esophagus between 3D-CT assisted with ABC and 4D-CT.

Objective To investigate the differences in position and volume between planning target volumes (PTV) based on positron emission tomography?computed tomography (PET?CT) images with an standardized uptake value ( SUV) no less than 2?5, 20% of the maximum SUV ( SUVmax ), or 25% of SUVmax , three?dimensional ( 3D ) CT, and four?dimensional ( 4D ) CT in thoracic esophageal cancer. Methods Eighteen patients with thoracic esophageal cancer sequentially received chest 3DCT, 4DCT, and [18F]fluoro?2?deoxy?D?glucose (FDG) PET?CT scans. PTV3D was obtained by conventional expansion of 3DCT images;PTV4D was obtained by fusion of target volumes from 10 phases of 4DCT images. The internal gross tumor volumes ( IGTV) , IGTVPET2.5 , IGTVPET20%, and IGTVPET25%, were generated based on PET?CT images with an SUV no less than 2?5, 20% of SUVmax , and 25% of SUVmax , respectively. These IGTVs were expanded longitudinally by 3?5 cm and radically by 1 cm to make PTVPET2.5 , PTVPET20%, and PTVPET25%, respectively. Results PTV3D was significantly larger than both PTV4D and PTVPET(P=0?000 -0?044), while there was no significant difference between PTV4D and PTVPET ( P= 0?216 -0?633 ) . The mutual degrees of inclusion ( DIs ) between PTV3D and PTV4D were 0?70 and 0?95, respectively, which were negatively correlated with 3D?Vector ( P=0?039). The mutual DIs between PTVPET2.5, PTVPET20%, and PTVPET25% were 0?74, 0?72, 0?78, 0?73, 0?77, and 0?70, respectively, which showed no correlation with 3D?Vector (P=0?150 -0?822). The mutual DIs between PTV3D and PTVPET were 0?86, 0?84, 0?88, 0?63, 0?67, and 0?59, respectively. Conclusions It is difficult to achieve complete volumetric overlap of PTVs based on 3DCT, 4DCT and PET?CT in thoracic esophageal cancer due to different target volume information. PET scan during free breathing should be used with caution to generate PTVs in thoracic esophageal cancer.