Objective To investigate the significance of the expression of endostatin and vascular endothelial growth factor(VEGF)in hepatocellular carcinoma(HCC).Methods The expression of endostatin and VEGF in 46 specimens which were collected at Second Affiliated Hospital of Fnjian Medical University from January 2000 to Apfil 2005 was detected by immunohistochemistry.The relationship between the expression of endostatin and VEGF and the progression of HCC was determined.All data were analyzed via t test,paired t test,Pearson rank correlation coefficient.LSD-t test or Tamhane's-t test.Results The expression of endostatin and VEGF was detected mainly in the cytoplasm of HCC cells and adjacent tissues.The values of mean optical density (MOD)of endostatin in HCC tissue,adjacent tissue and normal liver tissue were 0.11±0.02,0.14±0.0l and 0.09±0.01.respectively,and the values of integrated optical density(IOD)of endostatin in the above mentioned tissues were(1.8±0,2)×10~4,(3.8±2.2)×10~4 and(0.9±0.4)×10~4,respectively.The values of MOD and IOD of endostatin in HCC tissue were significantly lower than those in adjacent tissue(t=2.032,7.927,P＜0.05).The values of MOD of VEGF in HCC tissue,adjacent tissue and normal liver tissue were 0.13±0.02.0.12±0.02 and 0.11±0.02,respectively,and the values of IOD of VEGF were(5.4±3.1)×104,(3.9±2.5)×10~4 and(3.0 ±3.0)×10~4,respectively.The values of MOD and IOD of VEGF in HCC tissue were significantly hisher than those in adjacent tissue(t=5.871,8.723,P<0.05).There was positive correlation between the expression of endostatin and the recurrence of HCC(r=0.669,P<0.05),while VEGF had no influence on the recurrence of HCC(t=0.892,P>0.05).Conclusions Endostatin is mainly expressed in HCC tissue,but the level of its expression is lower in HCC tissue than in adjacent tissue.The expression of VEGF is higher in HCC tissue than in adjaneet tissue.Endostatin,rather than VEGF,may be used as a independent prognostic factor of the prognosis of patients with HCC.

Objective To study the distribution of IL-6 gene promoter 572C/G, 634C/G polymorphisms in Han population of Guangxi province, and analyze the relation between IL-6 gene polymorphisms and plasma lipid, platelet count. Methods Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) was used to detect IL-6 genotype in 198 healthy Han adults. At the same time the plasma lipid level and platelet count were determined by routine methods. Results Serum lipid levels had not significant difference among the different genotypes of IL-6 (P

Objective To investigate the effects of dihydrofolate reductase (DHFR) gene expression up-regulating on cisplatin resistance in epithelial ovarian cancer cell lines.Methods The cDNA length of DHFR gene was amplified by PCR and was connected to lentiviral vector pWPI, the recombinant retroviral vector DHFR-pWPI was infected SKOV3 cells by lipofectamine 2000.The groups included DHFR-pWPI-SKOV3 cell, pWPI-SKOV3 cell and SKOV3 cell group.Western blot was used to detect the expression of DHFR.Flow cytometry was applied to measure the cell apoptosis rate of 3 groups cells in different cisplatin concentrations (2.5, 5.0, 10.0, 20.0 μg/ml) and at different time period (24, 48 and 72 hours), and half maximal inhibitory concentration (IC50) treated with cisplatin concentration (6.0, 4.0, 4.9 μg/ml).High performance liquid chromatography (HPLC) was applied to test intracellular cisplatin concentration in different cisplatin concentration (4.0, 6.0, 8.0 μg/ml) at 24 and 48 hours.Transmission electron microscope was used to observe ultrastructural changes cells under IC50 cisplatin concentration.Results The recombinant plasmid DHFR-pWPI was constructed and then infected into SKOV3 cell successfully.(1) The expression of DHFR detected by western blot in transfection group was higher than those in the negative control group and blank control group (10.280±0.009 vs 2.050±0.003 vs 3.480±0.003;P＜0.01).(2) Treated with cisplatin concentration (2.5, 5.0, 10.0, 20.0 μg/ml) at 24, 48 hours, the apoptosis rate detected by flow cytometry results shown that they were lower than those in the negative control group and blank control group (P＜0.05), while treated at the concentration of 5.0 and 10.0 μg/ml for 72 hours, whose apoptosis rate in transfection group was higher than those in the negative control group and blank control group (P＜0.05).When treated cells under IC50 cisplatin concentration (6.0, 4.0, 4.9 μg/ml) at for 24 and 48 hours, the results indicated thatthere were mainly G0/G1 stage cell cycle rate in 3 groups, it was obviously higher in transfection group than those in two control groups (P＜0.05).However, mainly G2/M, S stage cell cycle rate for 72 hours, and S stage cell cycle rate in transfection group obviously higher than those in two control groups, but G2/M stage cell cycle rate were lower (P＜0.01).(3) After treated with cisplatin concentration (4.0 μg/ml) for 24, 48 hours and cisplatin concentration (6.0 μg/ml) for 24 hours, the intracellular cisplatin content tested by HPLC method in the transfection group were significantly lower than those in two control groups (P＜0.01).While, at 6.0 μg/ml of cisplatin concentration for 48 hours and 8.0 μg/ml of cisplatin concentration for 24 and 48 hours, the intracellular cisplatin content of transfection group were obviously higher than those two control groups (P＜0.05).(4) Treated with IC50 (6.0, 4.0, 4.9 μg/ml) cisplatin concentration at different time to obeserve ultrastructural changes by transmission electron microscopy.The results shown that the microwire gathered together at 24 and 48 hours, and the number and structure of mitochondria had obvious change in transfection group, while there was rare microfilament, the number of mitochondria decreased but structure change was not apparent in two control groups.There were appeared expansion of endoplasmic reticulum and had rare normal organelles among three groups.After treated with cisplatin for 72 hours, there were inordinate microfilament, a part of nuclear membrane disappeared, a lot of ribosomes gathered together in two control groups, and there were rare microfilament in transfection group, nuclear membrane completely disappeared, many white cystic matter were seen in cytoplasm, mitochondrial structure disappeared completely, which seems most cells on the verge of death.Conclusion The lentiviral expressing vector harboring human DHFR gene were successfully constructed.When the up-expression of DHFR gene, the drug-resistant in ovarian cancer cell may be increase, which suggest that there were certain contact between resistance increases with microfilament gathered and the change of the mitochondria.

Background and purpose:Dihydrofolate reductase (DHFR) is expressed highly in platinum-resis-tant ovarian cancer. This study aimed to explore the relationship between the silence ofDHFR gene and platinum drug resistance in ovarian cancer, and lay the foundation for the treatment of platinum-resistant ovarian cancer.Methods:To design targeting hairpin siRNA ofDHFR gene, the optimal siRNA silent sequence was selected, and lentiviral vector carryingDHFR gene was constructed successfully, named DHFR-pGCSIL-SKOV3 cell. Flow cytometry was used to detect the cell apoptosis of DHFR-pGCSIL-SKOV3 cells, pGCSIL-SKOV3 cells and SKOV3 cells incubated in various concentrations of cisplatin (2.5, 5.0, 10.0 and 20.0 μg/mL) at different time points (24, 48 and 72 h), and cell cycle changes of these cells at IC50 cisplatin concentration (4.4 μg/mL). High performance liquid chromatography was used to test intracellular concentration of cisplatin at different induction concentration of cisplatin (2.5, 5.0 and 7.5 μg/mL) and various time points (24 and 48 h). Ultrastructural changes of these cells at concentration of cisplatin IC50 (4.4 μg/mL) were observed by transmission electron microscope.Results:After annealing double-strand nucleotide was connected to pGCSIL/GFP vector, sequencing result was correct. SKOV3 cell were transfected with virus particles followed by Western blot detection of interference effect. Flow cytometry was used to detect apoptosis in three groups of cells, and increased apoptosis rate was found at the raised cisplatin concentration (2.5, 5.0, 10.0 and 20.0 μg/mL) at 24, 48 and 72 h in DHFR-pGCSIL-SKOV3, pGCSIL-SKOV3 and SKOV3 cells. The apoptosis rate in DHFR-pGCSIL-SKOV3 was signiifcantly higher than that in pGCSIL-SKOV3 and SKOV3 cells at 24 and 48 h (P<0.05). Flow cytometry was adopted to test cells cycle of 3 groups at different time period under IC50 cisplatin concentration (4.4 μg/mL), the results indicated that G0/G1 phase cell rate of DHFR-pGCSIL-SKOV3 was much more than the others, of which G2/M and S phase cell rates were on the contrary. While at 72 h, 3 groups were mainly G2/M and S phase cell rates, DHFR-pGC-SIL-SKOV3 was lower than the others. High performance liquid chromatography method was used to detect intracellu-lar cisplatin concentration at 24 and 48 h after the cells were incubated at various concentrations of cisplatin (2.5 and 5.0μg/mL). The results showed the intracellular cisplatin content of DHFR-pGCSIL-SKOV3 cell was signiifcantly higher than that of pGCSIL-SKOV3 and SKOV3 cells. However, after incubation at cisplatin concentration of 7.5 μg/mL, the intracellular cisplatin content of DHFR-pGCSIL-SKOV3 cell was signiifcantly lower than that of pGCSIL-SKOV3 and SKOV3 cells at 24 h, while higher than pGCSIL-SKOV3 and SKOV3 cells at 48 h (P=0.034,P=0.014). We observed ultrastructural changes of three different cell lines induced by IC50 cisplatin concentration(4.4 μg/mL) at different time points by the electron microscope. We found that the microiflaments were increased and gathered together and mitochondrial structure was also changed obviously without the drug. However, there was rare microiflament in three groups of cells at 24 and 48 h, while at 72 h, obviously increased inordinate microiflaments were observed.Conclusion:We successfully constructed pGCSIL lentivirus interference carrier carryingDHFR gene. The research indicates that down-regulation ofDHFR gene is related to cisplatin drug resistance in ovarian cancer. The results laid the foundation for us to investigate the molecular mechanisms of multidrug-resistance in tumor.

Objective To explore the mid to long term therapeutic efficacy of Graves′ disease after interventional embolization. Methods Twenty five patients of Graves′ disease treated with interventional embolization were followed up for 24-57 months. T 3 and T 4 were monitored at pre operation, six months, 12 months, 2, 3, and 4 years after operation, respectively. Other references included pulse, thyroid size, and vessel′s murmur. Results Twenty two patients completely relieved from the hyperthyroidism during the follow up. Only one patient suffered from recurrence. Other two patients were still on maintaining dosage of antithyroid drug therapy. No hypothyroidism or hypoparathyroidism was found during this term. Conclusion Mid to long term follow up showed satisfactory efficacy of interventional therapy, offering another alternative for treatment of refractory Graves′ disease.

Objective Stress effect plays an important role in the development of some myocardial diseases. We hypothesized it was important nosogenesis to myocardial damage and cardiac allograft vasculopathy. Methods The transplanted hearts from Lewis to Wister rats served as allografts and from Lewis to Lewis rats as isografts based Ono's model. The differential proteins in the transplanted hearts were separated by comparative proteome, and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and searched by Matrix Science software system.Results All transplanted hearts were characterized by lumen loss [(total vessel area-luminal area)/total vessel area] in the coronary artery 2 weeks after the operation [(2.07%±0.93%) vs. (27.58%±11.14%), P＜0.01], but more predominant after 8 weeks [(2.34%±1.06%) vs. (72.29%±20.57%), P＜0.01]. All samples of the left ventricle were analyzed by proteomic techniques and 37 distinct proteins involving their respective isoforms and subunits were identified. Nine proteins were correlated to endoplasmic reticulum stress effect and myocardial damage, and 2 proteins were verified by Western blot.Conclusion Stress plays an important role in cardiac allograft damage and the development of rat cardiac allograft vasculopathy.

Objective To determine whether autologous skeletal myoblasts implantation improves the cardiac function after myocardial infarction and the possible mechanism. Methods Myocardial infarction was induced by ligation of the left anterior descending coronary artery in rabbits. At 2 weeks, 1. 34 × 107 to 1.75 × 107 autologous skeletal myoblasts were infused into the lesion via direct intramuscular injection. In the control group, the postinfarction hearts were infused with medium alone. Buxco invasive cardiac function testing and histopathological examination were utilized to evaluate the functional and structural changes in the myocardium 4 weeks later. Results Both maximum rising rate of the left intraventricular pressure [+dp/dtmax,( 1 217.77 +89.91 )mmHg/s vs. (897.83 ±70.04) mmHg/s] and maximum falling rate of the left intraventricular pressure [- dp/dtmax,( -1174.58 ± 91.5 ) mmHg/s vs. ( - 753.67 ± 69.66 ) mmHg/s] were improved in the myoblast transplanted group compared with medium infusion group. The positive desmin immunostaining skeletal myofibers in the myocardium were found throughout the infracted areas and the border zone. Conclusion Autologous skeletal myoblasts can establish muscle tissue when transplanted into postinfarction hearts, and this mucle can treat myocardiac infarction effectively.

Background As assessment of the peripapillary retinal nerve fiber layer (RNFL) has been an important approach for detecting structural damage in patients with glaucoma and myopia is a vital risk factor of primary open glaucoma,it is urgent to establish the correlation between RNFL thickness and myopia,not only for understanding the characteristics of RNFL with the change of the degree of myopia,but also for identifying those myopic patients with the early stage of glaucoma.Objective This study was to assess the influence of myopia for the thickness of RNFL measured by 3D optical coherence tomography (3D-OCT).Methods Two hundred and fifty-eight eyes of 258 myopic subjects from General Hospital of Ningxia Medical University were recruited.The myopic eyes were divided into low myopia group (42 eyes,-0.5 D ≤ SE ≤-3.0 D),middle myopia group (120 eyes,-3.0 D＜SE≤-6.0 D),high myopia group (58 eyes,-6.0 D＜SE≤-8.0 D) and extreme high myopia group (38 eyes,SE ＞-8.0 D).The peripapillary RNFL thickness profile including temporal,superior,nasal and inferior quadrants and each of the 12 clocks was measured by 3D-OCT.The measured values were compared among different degrees of myopia,and the correlations between spherical equivalent (SE) and axial length with RNFL thickness were analyzed using linear regression equation.Results The RNFL thickness was gradually declined with the increase of SE and elongation of axis,showing significant differences among the 4 groups in the superior,nasal and inferior quadrants and mean RNFL thickness (F=10.48,15.60,3.31,8.98,all at P＜0.05),but temporal RNFL thickness was increased with the SE rise,with markedly difference among the 4 groups (F =2.92,P =0.03) ; and RNFL thicknesses in the superior,nasal,inferior quadrants and mean RNFL thickness were evidently declined in the high and extreme high myopia group in comparison with low myopia group (all at P＜0.05).The overall RNFL parameters at 1:00,2:00,3:00,4:00,5:00,6:00,8:00,12:00 o'clock sectors were thinning as the increase of SE (all at P＜0.05) and unchanged at the 7:00,9:00,10:00,11:00 sectors in different SE groups (all at P＞ 0.05).Negative correlations were found between axial length or SE with the RNFL thicknesses at superior,nasal and inferior quadrants,average thickness as well as 1:00,2:00,3:00,4:00,5:00,6:00,11:00,12:00 o 'clock,and positive correlation was seen between the axial length or SE with the RNFL thicknesses at temporal quadrant.Conclusions The thickness of RNFL varys with the different degree of myopia and axial length.

Autophagy is known as cellular degradation of long-lived proteins and organelles, by a double-membranound structure fusing with the lysosome, for recycling intraceUular constituents and reusing nutri-tion. Defective autophagy is closely correlated with tumorigenesis and tumor progress. There are crosstalks between autophay associated/regulative genes and oncogene/anti-oncogene. More over, regulative autophagic cell death may lead to an novol way for cancer therapy.

BACKGROUND: By now, safety of collagenase application is still controvertible and some scholars believed that collagenase might induce the peripheral tissue injury. It attracts much attention in clinic that whether there is nerve injury induced by collagenase chemonucleolysis (CCN) around the injection sites.OBJECTIVE: To observe the effects of collagenase on spinal nerve conduction velocity (NCV) of rats detected with evoked potential method so as to probe into the safetyof collagenase application and further demonstrate the safety of percutaneous intervertebral disc CCN. DESIGN: Randomly grouping design, animal experiment.SETTING: Department of Interventional Radiology, Affiliated First Hospital of Sun Yat-sen University. MATERIALS: The experiment was conducted in the Basic Medical College of Sun Yat-son University from July to September 2002. A total of 57 male SD rats were randomly divided into normal group (n=9), acute sham-operation group (n=10), subacute sham-operation group (n=8), chronic sham-operation group (n=7), acute experimental model group (n=9), subacute experimental model group (n=7) and chronic experimental model group (n=7). METHODS: After being anaesthetized by intraperitoneal injection of continal (45 mg/kg) to separate and identify dorsal root ganglion (DRG),rats in the experimental group were locally dripped with 1 mL of collagenase (300 U/ML) and those in the sham operation group locally dripped with 1 mL of normal saline. Stimulating electrode was placed in the A point of sciatic nerve and recording electrode in the B point of ganglionic central process segment of DRG in L5 nerve root. Evoked potentials A and B were simulated to continuously record latency twice, and the average value was calculated; Distance between A and B were measured and recorded. NCV = distance between A and B / latency. Evoked potential of a segment ofnerve including DRG was measured in the acute group at one hour after administration, in the sub acute group at one week after administration and in the chronic group at one month after administration. MAIN OUTCOME MEASURES: NCV of each group. RESULTS: A total of 57 enrolled animals were involved in the analysis. NCV in normal group, acute experimental group, acute sham-operation group, subacute experimental group, subacute sham-operation group, chronic experimental group, chronic sham-operation group were (45.4±10.7), (43.4±5.9), (46.3±6.5), (52.4±10.4), (49.7±8.1), (46.7±11.0) and (44.6±6.5) m/s respectively. There were no remarkable differences in NCV among all the groups by using one-way analysis of variance (F=1.010,P=0.430); It was showed that there were no marked differences in NCV between each two groups by using multiple comparisons (P=0.336). CONCLUSION: Collagenase at the therapeutic concentration applied in clinical CCN has no remarkable effects on NCV of rat spinal nerve, and to a certain extent, percutaneous intervertebral disc CCN is relatively reliable.