Objective To detect the atherosclerotic progression in low density lipoprotein receptor (LDL-R) gene knock-out mouse by ultrasound biomicroscopy(UBM) technique,and to monitor the intima media thickness (IMT) and changes in plague of aortic wall.Methods 10 male LDL-R gene knock-out mice of 16 weeks age and 10 LDL-R gene knock-out mice of 24 weeks age were in the experimental group,and 10 male C57BL/6 mice of 16 and 24 weeks age were in the control group.The shapes of their aortic roots,ascending aorta,aortic arch and CCA were detected by UBM,and the IMT at aortic root view and carotid artery bifurcation were measured,then the data were compared with histopathology of the corresponding vascular segments.Results The difference between the IMT of aortic root and carotid artery bifurcation of the 16 week old LDL-R mice and the control group of the same age had no statistical significance.The difference between the IMT of carotid artery bifurcation of the 24-week-old LDL R mice and the control group of the same age had no statistical significance.The IMT of aortic root thickened compared with control group of the same age(P ＜0.01).Conclusions The UBM technique can be used to detect the atherosclerotic progression in LDL-R gene knock-out mouse.

Objective To investigate the expression of osteopontin and matrix metalloproteinase-9 (MMP9),and the relationship of osteopontin and MMP9 in malignant melanoma.Methods Expression of osteopontin and MMP9 was measured by immunohistochemical SP method in 23 patients with primary cuta- neous malignant melanoma,17 patients with metastatic melanoma and 20 patients with pigmented nevus. Results Osteopontin and MMP9 were expressed respectively in 87.5% and 75.0% of 40 malignant melanoma specimens,15.0% and 10.0% of 20 pigmented nevus specimens.The expression of both osteo- pontin and MMP9 was significantly higher (both P＜0.05) in malignant melanoma than in pigmented ne- vus.There was no correlation between the expression of osteopontin and MMP9,with age,sex,lymph node metastasis or location of lesions (P＞0.05).Twenty-nine cases were positive for both osteopontin and MMP9,4 negative for either osteopontin or MMP9.Conclusion Both osteopontin and MMP9 were over- expressed in malignant melanoma,but neither was related to lymph node metastasis.

Objective To study the clinical outcomes of titanium-coated interbody fusion system in treating lumbar isthmic spondylolisthesis.Methods A retrospective study was made in 22 cases with lumbar isthmic spondylolisthesis including 15 with degree I spondylolisthesis and seven with degree II spondylolisthesis treated by posterior lumbar interbody fusion using titanium-coated fusion cages from June 2002 to December 2003 in our hospital.There were six males and 16 females,with age range of 41-68 years(mean 52.5 years).In all cases,double cages were implanted with additional posterolateral bone graft,supplemented with pedicle screw systems.The average preoperative and postoperative disc space height,foraminal height,segmental and lumbar lordosis,spondylolisthesis and domain of sagittal rotation on flexion-extension radiographs were measured and fusion state observed.The Oswestry disability index (ODI)was used to evaluate the clinical outcome.Results The fusion rate was 95.4%.There was significant recovery of disc space height,foraminal height and domain of sagittal rotation.ODI decreased significantly.Conclusion The anatomic titanium-coated fusion system is effective and safe in treatment of lumbar isthmic spondylolisthesis because it has advantages in restoring sagittal alignment and keeping bony endplate intact.

To analyze the causes and treatments of high intraocular pressure ( > 21mmHg ) of angle - closure glaucoma underwent compound trabeculectomy 1mo after surgery.
●METHODS: This was a retrospective study of our hospital, from March 2010 to March 2013. Thirty-four (38 eyes) of angle-closure glaucoma patients were collected, who underwent compound trabeculectomy with high intraocular pressure ( > 21mmHg) 1mo after operation. We analyzed the causes and summarized the treatments.
●RESULTS:The causes which lead to early postoperative high intraocular pressure included malignant glaucoma (9 eyes of 8 cases), blood clot and connective tissue block under the scleral flap (15 eyes of 13 cases), hyphema after surgery(5 eyes of 5 cases), sclera flap incision was incarcerated with iris tissue ( 3 eyes of 3 cases ), preoperative high intraocular pressure for a long time(5 eyes of 4 cases), 1 eye of 1 case for unknown reason. After proper treatments, intraocular pressures of all patients were bellowed 21mmHg.
● CONCLUSlON: Early postoperative high intraocular pressure of angle - closure glaucoma underwent compound trabeculectomy is caused by various factors, and the early prevention and timely treatment are key points of a successful operation.

Objective:To investigate the expressions of miR-20a-5p and lysine (K) demethylase 6B (KDM6B) in osteosarcoma tissues and the effects of miR-20a-5p targeting KDM6B on the proliferation, migration and invasion of osteosarcoma cells and tumor growth.Methods:The clinicopathological and paracancerous tissues of 20 patients with osteosarcoma admitted to the First Affiliated Hospital of Chinese Medical University from January 2017 to March 2019 were collected. Quantitative real-time PCR (qRT-PCR) was used to detect the expression levels of miR-20a-5p and KDM6B mRNA in tissues. The osteosarcoma MG63 cells were divided into control group, mimic NC group, miR-20a-5p mimic group, and NC+ empty vector group, miR-20a-5p+ empty vector group, miR-20a-5p+ KDM6B group. The expression levels of miR-20a-5p and KDM6B mRNA of all groups were detected by qRT-PCR. Western blotting was used to detect the expression level of KDM6B. CCK-8 assay, cell scratch test and Transwell test were used to detect cell proliferation, migration and invasion ability. According to the random number table method, nude mice were divided into NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group, with 5 mice in each group. Tumor growth ability was detected by tumor xenograft nude mouse models.Results:The relative expression level of miR-20a-5p mRNA in osteosarcoma tissues was 0.55±0.27, and that in paracancerous tissues was 1.22±0.28, with a statistically significant difference ( t=7.701, P<0.001). The relative expression level of KDM6B mRNA in osteosarcoma tissues was 1.66±0.19, and that in paracancerous tissues was 1.00±0.15, with a statistically significant difference ( t=12.219, P<0.001). After transfection of miR-20a-5p, KDM6B mRNA and protein expression levels decreased with the increase of miR-20a-5p expression level. After miR-20a-5p transfection for 48 h, the cell proliferation abilities of the blank control group, mimic NC group and miR-20a-5p mimic group were 0.83±0.04, 0.81±0.03 and 0.52±0.01 ( F=89.655, P<0.001), compared with the blank control group and mimic NC group, the cell proliferation ability was significantly inhibited in the miR-20a-5p mimic group (both P<0.001). The cell proliferation abilities of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were 0.83±0.05, 0.52±0.01 and 0.67±0.05 ( F=43.919, P<0.001), compared with the NC+ empty vector group, the cell proliferation ability was significantly inhibited in the miR-20a-5p+ empty vector group ( P<0.001); compared with the miR-20a-5p+ empty vector group, the cell proliferation ability of miR-20a-5p+ KDM6B group increased significantly ( P<0.001). The scratch healing rates of the blank control group, mimic NC group and miR-20a-5p mimic group were (32.51±2.73)%, (30.26±3.22)% and (13.52±1.77)% ( F=46.314, P<0.001), compared with the control group and the mimic NC group, the scratch healing rate of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The scratch healing rates of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (31.34±3.11)%, (12.15±1.64)% and (28.93±2.89)% ( F=47.511, P<0.001), compared with the NC+ empty vector group, the scratch healing rate of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the scratch healing rate of miR-20a-5p+ KDM6B group was significantly increased ( P=0.001). The numbers of transmembrane cells in the blank control group, mimic NC group and miR-20a-5p mimic group were 114±16, 108±11 and 42±6 ( F=36.282, P<0.001), compared with the control group and mimic NC group, the number of transmembrane cells of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The numbers of transmembrane cells in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group was 143±11, 39±4 and 139±12 ( F=112.120, P<0.001), compared with the NC+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ KDM6B group was increased significantly ( P<0.001). The tumor volumes of mice for 21 d in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (1 667.50±250.40) mm 3, (129.20±21.00) mm 3 and (775.41±77.51) mm 3 respectively, with a statistically significant difference ( F=77.651, P<0.001). The tumor weights of the 3 groups were (1.35±0.18) g, (0.12±0.01) g and (0.61±0.03) g respectively, with a statistically significant difference ( F=104.191, P<0.001). Conclusion:The expression of miR-20a-5p is significantly decreased in osteosarcoma tissues, and the expression of KDM6B is significantly increased in osteosarcoma tissues. Overexpression of miR-20a-5p may inhibit the proliferation, migration and invasion of osteosarcoma cells and tumor growth by targeting to reduce the expression of KDM6B.

The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF.
Animals ; Cell Line ; Chickens ; Chorioallantoic Membrane ; blood supply ; Disease Models, Animal ; Genetic Therapy ; Humans ; Male ; Myocardial Infarction ; metabolism ; pathology ; therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

This study is to investigate the protective effect of longistyline A against corticosterone-induced neurotoxicity in PC12 cells. While PC12 cells were exposed to 100 micromol x L(-1) corticosterone for 48 h, cell survival rate was reduced and lactate dehydrogenase (LDH) release increased. In parallel, corticosterone caused significant elevations of DNA fragmentation, [Ca2+]i and caspase-3 activity. However, when the PC12 cells were incubated with longistyline A (4.0, 8.0 and 16.0 micromol x L(-1)) in the presence of 100 micromol x L(-1) corticosterone for 48 h, the effects were evidently alleviated, but dose-dependent manner was not obvious. In summary, longistyline A could generate a neuroprotective effect against corticosterone-induced neurotoxicity in PC12 cells possibly by decreasing [Ca2+]i and caspase-3 activity.
Animals ; Cajanus ; chemistry ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Corticosterone ; toxicity ; DNA Fragmentation ; drug effects ; L-Lactate Dehydrogenase ; metabolism ; Molecular Structure ; Neuroprotective Agents ; isolation & purification ; pharmacology ; PC12 Cells ; Phenols ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats

Drying is the critical link during pharmaceutical process of traditional Chinese medicine (TCM), which is directly related to the quality of drugs. The key to technology upgrading of pharmaceutical equipment in Chinese materia medica enterprise is the development of new drying techniques, which concerns the modernization of TCM. The study provides new ideas for the drying technology and equipment by means of reviewing the research status of drying process for the traditional Chinese medicinal materials and preparations, and analyzing the traditional and modern drying methods and equipment, as well as their existing problems and corresponding measures for the drying processes and equipment. In addition, this paper expounds the development trend of traditional Chinese medicinal materials and preparations of drying process and equipment.
Chemistry, Pharmaceutical ; instrumentation ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; Humans ; Medicine, Chinese Traditional ; instrumentation ; standards ; Plants, Medicinal ; chemistry

Objective To compare the effects of hepatic vein occlusion with tourniquet and Satinsky clamp in reseeting liver tumor involving the second hepatic portal.Methods From Jan 2003 to Jun 2006,180 patients underwent major liver resection with the selective hepatic vascular exclusion (SHVE).According to methods of hepatic vein occlusion,they were divided into two groups:Occlusion with tourniquet(tourniquet group,n=95)and occlusion with Satinsky clamp(Satinsky clamp group,n= 85).In tourniquet group,the hepatic veins were encircled and occluded with tourniquet,and in Satinsky clamp group,the hepatic veins were not encircled and clamped directly with Satinsky clamp.Data regarding the intraoperative and postoperative courses of the patients were analyzed.Results There was no difference between the two groups regarding the operating time,ischemia time,intraoperative blood loss and postoperative complications rate.The dissecting time of hepatic veins was significantly shorter in Satinsky group(6.2?2.4 min vs 18.3?6.2 min).lu the tourniquet group,five hepatic veins(one fight hepatic vein and four common trunk of left-middle hepatic veins)could not be dissected and encircled because of the tumors involving the cava hepatic junction.Another patient's common trunk of left-middle hepatic vein was inadvertently lacerated during the dissection.Hepatic veins in these 6 patients were occluded with Satinsky clamp successfully.Conclusion Occlusion with Satinsky clamping is safer and easier procedure than tourniquets in the resection of liver tumor involving the second porta hepatis.

OBJECTIVETo study the cytotoxicity induced by chrysotile asbestos (CA), rock wool (RW) and wollastonite (WS).

METHODSV79 cells were divided into 4 groups. i.e. CA group, WS group, RW group and control group (200 microl PBS). The exposure concentration of dusts was 100 mg/L, The cell viability was detected by MTT and lactate dehydrogenase (LDH) activity assays. The technique of scanning electron microscopy was used to examine the change of V79 cells.

RESULTSSiO2 was main constituent for 3 kinds of dusts. In MTT assay, the cell viability of RW and WS groups was 64.8% and 65.7%, respectively, which were significantly higher than that (54.5%) of CA group (P < 0.01). In LDH assay, the LDH activity of RW and WS groups [(15.7 +/- 50.9), (12.3 +/- 3.7) U/L, respectively] was significantly lower than that [(20.2 +/- 0.9) U/L] of CA group (P < 0.05). In scanning electron microscopy examination, it was found that the two ends of V79 cells in CA group contained a great deal of fibers remaining bodies, but the V79 cell appearance in RW and WS groups was normal.

CONCLUSIONThe cytotoxicity induced by RW and WS is significantly lower than that induced by CA for V79 cell.

Animals ; Asbestos, Serpentine ; toxicity ; Calcium Compounds ; toxicity ; Cell Line ; drug effects ; Cricetinae ; Cytotoxins ; toxicity ; Lactate Dehydrogenases ; metabolism ; Mineral Fibers ; toxicity ; Silicates ; toxicity